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. 2026 Jun 18;21(6):e0348280. doi: 10.1371/journal.pone.0348280

Intelligent automation cognitive system for accurate malaria diagnosis using digital blood smears

Emad Malaekah 1, Husham Saied 1, Othman Alfahad 1, Tatyana Utkina 2, Marwa A Saleh 3,*, Ahmad abduaziz Aljaffer 4
Editor: José Luiz Fernandes Vieira5
PMCID: PMC13278421  PMID: 42313826

Abstract

Malaria is a potentially fatal illness caused by a parasite of the genus Plasmodium that humans get by being bitten by female Anopheles mosquitoes carrying the infection. The incidence of malaria worldwide is disproportionately high in the African continent. Automated systems and cognitive analysis of digitized images of blood smears were used to diagnose Plasmodium malaria. This method is implemented in the Aidos intelligent system, which is easily accessible online. For the study, the database included images of 191 blood smears of patients infected with malaria and 227 images of blood samples from healthy patients. The images were digitized using the method developed by Professor Lutsenko E.V. The images were digitized for 12 light spectra. Then, spectral analysis of the blood smear images was carried out only for 18 new patients, and the duration was 10 seconds. The average similarity value of Plasmodium malaria recognition in patients was achieved at 66.965%. No false positive decisions were obtained for digitalized blood smears from healthy patients. The automated system-cognitive analysis of digitized blood smears provides instant diagnostic support. It allows medical workers with limited knowledge in microscopy and artificial intelligence to perform diagnostics.

1. Introduction

Humans contract malaria via the bites of female Anopheles mosquitoes carrying the tiny parasite called Plasmodium that causes the sickness. A person’s red blood cells are destroyed by the Plasmodium malaria parasite, which then replicates in other cells. Typical symptoms of malaria include fever, headache, muscle soreness, and exhaustion [1]. WHO estimates that there will be 282 million cases of malaria and 610,000 deaths in 2024 – about 9 million more than the previous year [2].

A correct diagnosis at an early stage will reduce the cost of malaria treatment. Plasmodium malaria is detected by examining a drop of a patient’s blood, spread as a “blood smear” on a microscope slide, under a microscope [3]. Modern imaging techniques make digitizing microscopic blood smears with high optical resolution possible. A decision support system that assists microscopists in malaria diagnosis is available using computer vision to screen digital thin blood films, but it is prone to false negative decisions [4]. To count the number of infected cells in a single blood thick film, a skilled microscopist usually takes 20–30 minutes to carefully examine the film. Plasmodium malaria screening becomes a lengthy and error-prone process when there are many patients, and manual blood film evaluation mostly depends on skilled staff. When evaluating treatment results and the effectiveness of antimalarial medications, the parasite clearance rate is crucial [5]. Rapid tests, which use a small kit to identify antigens from malaria parasites, can be used to identify malaria. Through a hole in the kit, a drop of blood is added, and the tests are conducted inside the apparatus, yielding findings quickly [6]. However, the rapid diagnostic tests (RDTs) is imprecise, and any incorrect result may impact patient treatment [7].

Automated methods can help many patients detect Plasmodium malaria quickly, cost-effectively, and accurately. Sophisticated image processing techniques using shape, color, intensity, size, and texture automate Plasmodium malaria detection [8,9]. In laboratories, peripheral blood smears are stained with Giemsa to improve the reliability of malaria diagnostics [10]. Giemsa staining allows one to detect morphological features present in certain types of samples and absent in others. Lasers with an energy of 80–95 nJ remove the blood plasma covering the infected blood cells of interest from blood smears [11]. Diagnostics for Plasmodium malaria should be accurate, identify parasites in areas where they are endemic, and produce fewer false negative results [12]. Oxford University’s Visual Geometry Group (VGG) created a convolutional neural network for large-scale image recognition [13]. There are systems for processing pictures of blood smears captured by a smartphone camera mounted on a microscope’s ocular [14,15]. Convolutional neural network (CNN)-based deep learning techniques for smartphones may identify malaria parasites in thick blood smears by extracting and categorizing features [16]. Images of disease-infected and uninfected blood cells are processed by CNN automatically, without the need for human assistance [17,18]. However, these methods and tools are not available for widespread use. Deep belief networks (DBNs), probabilistic generative models consisting of different layers of hidden variables with associations between layers, are used for malaria detection [19,20]. However, DBNs require a labor-intensive setup procedure to be applied. An imperative capsule neural network is used to detect malaria. However, processing data using this network takes more than 33 minutes [21]. The primary challenge in all published research utilizing artificial intelligence and digital microscopy is the absence of high-quality blood smears or thick films that can identify sparse low parasitemia [22,23].

For molecular diagnostics of malaria at the point of care, integrated point-of-care devices (Point-of-Care) are used. These are compact automated systems that detect DNA or RNA of Plasmodium parasites directly at the patient’s site (in a clinic, field hospital, or laboratory). They utilize molecular biology methods such as Polymerase Chain Reaction or Loop-Mediated Isothermal Amplification [24]. Although the devices are automated, humans still play a vital role in malaria diagnostics. A specialist collects a drop of blood from a finger or vein, places the blood in a cartridge, inserts the cartridge into the device, checks the expiration date of the reagents, and initiates the analysis. The physician must maintain sterile technique and consider the epidemiological situation, the patient’s symptoms, and the results of other tests.

Smoothing filters such as Gaussian, median, and geometric mean filters suppress noise in microscopic images [25]. Even though the use of computer diagnoses is expanding rapidly, there are still issues with identifying low malaria parasites in blood smear microscopic pictures [26].

For more than 30 years, Aidos, an open intellectual system, has been used with success. It can be applied to any subject where a scientist or practitioner uses the most recent advancements in artificial intelligence to solve difficulties in their line of work and continuously expand their expertise [27]. It is suggested that an automated system-cognitive (ASC) analysis, which is incorporated into the universal intellectual system Aidos and was created by Professor Lutsenko, be used to identify blood smears infected with malaria [28]. The Aidos system’s ASC analysis model is based on systemic fuzzy interval mathematics, which makes it possible to handle big, complicated datasets that are interconnected, noisy, and fragmented. It accepts data in a variety of units of measurement and measurement scales, including numeric, ordinal, and nominal [29]. The Aidos system is also used for intelligent image processing, i.e., for their digitization, creation of models of specific images, abstraction, identification, classification of generalized images and solving several other problems. as a result of generalization, the value of image features for their differentiation is revealed, as well as the degree of specificity of certain features for specific images [30]. This allows, without compromising the adequacy of the model, to remove low-value features from it, i.e., to carry out abstraction of generalized images, which subsequently ensures a reduction in the costs of various types of resources for collecting and processing graphic information.

The present work aims to develop an intelligent system that can provide effective preliminary diagnostics of malaria for use in areas with insufficient medical care.

2. Methods

2.1. Formalization of the subject area

To study complex systems, it is necessary to transform source data into information, and then into knowledge. This knowledge is used to solve problems of classification, decision support, and domain research by exploring its system-cognitive model, generating a large number of tabular and graphical output forms. The process of transforming source data into information, and then into knowledge, is shown in Fig 1. For this study, it is necessary to first convert the graphic data into a text file.

Fig 1. The process of transforming raw data into information and knowledge.

Fig 1

Images for malaria diagnosis obtained by Machine Learning Engineer Nagadia from AURO University (Surat, Gujarat, India) and published online [31] were used for malaria diagnostics. This dataset contains 550 images of malaria (Plasmodium)-positive and negative blood smears, arranged in two folders, Train and Test, which can be used for training and testing, respectively. In this study, images from the Train folder were used. Examples of malaria-negative blood smear images are shown in Fig 2, and examples of malaria-positive are shown in Fig 3.

Fig 2. Malaria Plasmodium-negative blood smear samples.

Fig 2

Fig 3. Malaria Plasmodium-positive blood smear samples.

Fig 3

To solve the problems in the article, we will use the software interface from the Aidos system to study images by their spectrum. To do this, download the Aidos system from the author’s website at the link: http://lc.kubagro.ru/aidos/_Aidos-X.htm and install it per the instructions on the website. Write the scanned images of blood smears to the folder  ..\AID_DATA\Inp_data\. A fragment of the structure of the original image files in JPG format is shown in Fig 4. The blood smears were not pre-processed and were used as they are posted in the Kaggle repository (https://www.kaggle.com/datasets/imdevskp/malaria-dataset). The blood smear photographs in the Kaggle repository dataset are publicly available, and the patient names are not provided, so no ethical approval or permission to use the samples for research is required.

Fig 4. Structure of source image files.

Fig 4

Mode 2.3.2.5 of the Aidos intelligent system was used to digitize the images. The appearance of this image processing mode is shown in Fig 5. This mode provides the conversion of images in BMP or JPG format into the lnp_data.xlsx source data file, in which a line represents each image. The subject area is formalized, models are created, and tested using this source data file.

Fig 5. Interface of image processing mode by pixels and spectrum.

Fig 5

The image digitization method developed by Professor E.V. Lutsenko [32] was used in this study. A patent protects the essence of the image digitization method by the intelligent Aidos system [33]. The method of digitizing images into a numerical feature vector proposed by Professor Lutsenko has the following features: the object’s contour is used; the image is converted to a polar coordinate system; a radial shape function is constructed; the obtained parameters are used as descriptors for image comparison. A typical algorithm (script) of the method consists of the following steps: scanning or photographing an object; converting the image to a standard format; performing object segmentation; obtaining a contour image. To convert the image to a standard format, the following are performed: centering the object, scaling it, and rotating it to a standard position. This makes the analysis independent of image orientation. The image center is taken as the origin of coordinates. For each pixel, its radial function is calculated, which determines: the average radius, dispersion, spectral characteristics, shape harmonics, and derivatives of the radial function. The resulting feature vector is used for classification, clustering, object identification, and constructing shape prototypes.

The Aidos system detects the colors in a picture with high accuracy by measuring the spectra of graphic objects. The picture spectrum is defined by the system as the percentage of pixels of various hues in the total number of pixels in the image. To measure the image spectra, the number of color ranges is specified. The Aidos system calculates the brightness of the Red, Green, and Blue rays to create a spectrum, and each color range represents a mix of these rays. The spectra of particular items are then compared with those of training classes. In this instance, the entire amount of information about an object’s color spectrum that pertains to its inclusion in the class’s generalized picture is computed. Digitizing 418 blood smear images was labor-intensive and took 1 hour 25 minutes on a computer with an AMD Athlon 3.1 GHz 32-bit processor and Windows 10.

Table 1 shows a fragment of the file  .. \AID_DATA\Inp_data\inp_data.xlsx with the results of digitizing images for twelve spectra. Column A shows the number of the objects of study, i.e., blood smears. Column B shows the codes of the analyses that correspond to the names of the files with the images of blood smears. When conducting a study in the Aidos system, the data in this column is not used, but it can be used for visual control of the correctness of the identification of objects.

Table 1. Fragment of a file with the results of digitizing images of blood smears.

Object Analysis code Class SPECTRINTERV: 1/12 SPECTRINTERV: 2/12
A B C D E
No1 C133P94ThinF_IMG_20151004_155721_cell_111 Parasite 0.0000 0.0000
No2 C133P94ThinF_IMG_20151004_155721_cell_112 Parasite 0.0000 0.0825
No3 C133P94ThinF_IMG_20151004_155721_cell_113 Parasite 0.0000 0.0000
No4 C133P94ThinF_IMG_20151004_155721_cell_114 Parasite 0.0000 0.0000
No5 C133P94ThinF_IMG_20151004_155721_cell_115 Parasite 0.0000 0.6708

Column C is a classification and contains information about whether the patient is infected with malaria or not. The columns starting with column D are descriptive and contain digitized characteristics of the spectra of blood smear images.

2.2.  Synthesis of statistical and system-cognitive models

A formalized cognitive concept created by Professor E. V. Lutsenko enables the creation of mathematical models of the field under study. Conducting experiments reveals connections between elements: some elements are observed frequently, while others are rarely found together. Stable connections between elements suggest that they reflect an unavoidable reality, integral to these elements [34].

Mathematical models of the Aidos system are based on a matrix of absolute frequencies, reflecting the total counts, indicating the frequency of occurrences where levels of descriptive scales intersect with levels of classification scales (observations). But instead of using this matrix directly to solve every problem, matrices of conditional and unconditional percentage distributions are used. Calculated systemic cognitive models represent the data from experiencing a certain descriptive scale gradation, causing the modeling object to enter a state that corresponds to a specific classification scale (class) gradation. The models that are employed are A. Kharkevich’s measure of the amount of information, Karl Pearson’s χ-square, and the return on investment (ROI) coefficient, which is used in economics to manage an investment portfolio. The main idea behind these approaches is that the modeling object will be put into a particular state that corresponds to the class based on the quantity of information in the factor value. This makes it possible to process heterogeneous information about the states of the modeling object comparably and accurately [35].

Directly based on the empirical data, the matrix of absolute frequencies is calculated and presented in Table 2. Based on this, the matrix of conditional and unconditional percentage distributions is calculated and presented in Table 3. Then, based on Table 3, the matrices of systemic-cognitive models are calculated and presented in Table 4.

Table 2. Matrix of absolute frequencies (ABS statistical model).

Classes Sum
1 ... j ... W
Factor values 1 N11 N1jw N1W
...
i Ni1 Nij NiW NiΣ=∑j=1WNij\hspace{0.33em}
...
M NM1 NMj NMW
Total quantity
signs by class
NΣj=∑i=1MNij\hspace{0.33em} NΣΣ=∑i=1W∑j=1MNij
Total quantity
learning objects
samples by class
NΣj NΣΣ=∑j=1WNΣj\hspace{0.33em}

Table 3. Matrix of conditional and unconditional percentage distributions (statistical models PRC1 and PRC2).

Classes Unconditional
probability
sign
1 ... j ... W
Factor values 1 P11 P1j P1W
...
i Pi1 Pij=NijNΣj PiW PiΣ=NiΣNΣΣ
...
M PM1 PMj PMW
Unconditional
probability
class
PΣj

Table 4. System cognitive model matrix.

Classes Significance
factor a
1 ... j ... W
Factor values 1 I11 I1j I1W σ1Σ=1W−1∑j=1W(I1j−I―1)22
...
i Ii1 Iij IiW σiΣ=1W−1∑j=1W(Iij−I―i)22
...
M IM1 IMj IMW σMΣ=1W−1∑j=1W(IMj−I―M)22
Degree
reduction
class
σΣ1 σΣj σΣW H=1(W·M−1)∑j=1W∑i=1M(Iij−I―)22

Designations in tables:

i – value of the past parameter;

j – value of the future parameter;

Nij – number of meetings of the j-th value of the future parameter with the i-th value of the past parameter;

M – total number of values of all past parameters;

W – total number of values of all future parameters.

Ni – number of occurrences of the i-th value of the past parameter throughout the sample;

Nj – number of occurrences of the j-th value of the future parameter throughout the sample;

N – number of occurrences of the j-th value of the future parameter with the i-th value of the past parameter throughout the sample.

Iij – a particular knowledge criterion: the amount of knowledge in the fact of observing the i-th value of the past parameter that the object will go into the state corresponding to the j-th value of the future parameter;

Pi – unconditional relative frequency of meeting the i-th value of the past parameter in the training sample;

Pij – the conditional relative frequency of meeting the i-th value of the past parameter at the j-th value of the future parameter.

For the INF3 model, which is most often the most reliable, the following partial χ-square criterion is used – the difference between the actual and theoretically expected absolute frequencies:

Iij=Nij−NiNjN.

Identification-related problems like classification, recognition, diagnosis, and prediction, as well as decision-making assistance and subject-domain exploration through examination of its system-cognitive representation, are successfully handled by using system-cognitive models [36]. Three statistical models and seven knowledge models can be used concurrently in the Aidos system, which aids in resolving issues with identification, decision-making, and subject-area study based on two essential criteria. The Aidos system assesses how well different integral and private criteria work to solve these issues. The absolute frequency matrix shown in Fig 6 was computed directly from the data collected during the digitalization of blood smear images. It served as the foundation for the calculation of the conditional and unconditional percentage distribution matrices, which are shown in Figs 7 and 8, respectively.

Fig 6. Matrix of absolute frequencies.

Fig 6

Fig 7. Matrix of conditional percentage distributions.

Fig 7

Fig 8. Matrix of unconditional percentage distributions.

Fig 8

Typically, the acquired images have been contaminated by a variety of sounds, such as those caused by varying camera angles and microscope settings. To detect and reduce noise in the original data, as well as to identify disinformation in the original data and restore the real information, software tools are required. The training sample item is deemed atypical (an artifact) if the recognition results show that its degree of similarity with the class is below the desired threshold. There is a method for identifying, eliminating, and adding artifacts in the Aidos system. The percentage represents the threshold degree of similarity [37]. Fig 9 shows the model’s degrees of adequacy after removing artifacts with recognition accuracy below 25%. The L1 measure for the INF3 model, an information/knowledge model used to build “information portraits” and compute integrated similarity measures. Then equaled 0.976, which is a satisfactory result for medical intelligent applications.

Fig 9. Generalized form of estimation of model reliability.

Fig 9

The database includes digitized data of 191 blood samples of patients infected with malaria and 227 blood samples of healthy patients, and unrecognized digitized data were excluded from the database. To obtain an alternative solution, the virtual patient features equal to 45.0000 were added to the database containing digitized values of patient blood samples. If the database includes blood samples of only infected or only uninfected patients, the Aidos system will not be able to recognize them.

3. Results and discussion

3.1. Results of recognition of the examined blood smears

Using the INF3 model, digitalized blood smear image data were recognized with a high degree of similarity; examples of malaria recognition are shown in Fig 10. For several patients, a malaria recognition accuracy rate of 100% was achieved.

Fig 10. Visualization of malaria recognition results.

Fig 10

The diagnostic accuracy for detecting malaria-infected blood smears was 100%. All infected samples were correctly recognized at varying similarity levels. Fig 11 presents a pie chart illustrating these recognition results: different colors represent different similarity ranges for malaria-positive samples. Notably, 31.9% of infected records were identified at a similarity threshold of 80%. Across the full database of malaria cases, diagnostic accuracy remained 100%, with an average similarity score of 66.97% compared with the reference “Parasite” class. In the right-hand column, color-coded bars display the achieved similarity levels for each detected malaria case.

Fig 11. Diagrams of malaria blood sample recognition results.

Fig 11

Legend shows the levels of malaria detected in different colors. A fragment of the recognition of digitized blood smears of healthy patients is shown in Fig 12. The obtained similarity level is low. Still, the most important thing is that these samples are not classified as false positive solutions. The results of recognizing the absence of malaria in healthy patients are lower than those of known methods [17,18], for using the intelligent Aidos system, there is no need to purchase a license; it is available on the Internet and is recommended for use in developing countries.

Fig 12. Results of recognition of digitalized blood smears of healthy patients.

Fig 12

3.2.  Results of the selection of light spectra for the study

The Aidos system allows for the determination of the light spectra that can detect malaria. The result of this spectrum is shown in Fig 13.

Fig 13. The importance of light spectra for detecting malaria.

Fig 13

Fig 14 shows the Pareto curve of the significance of descriptive scales (features) [38]. 7% of the most significant features provide 50% of the total importance. Half (50%) of the most considerable features provide 96% of the total importance. The number of gradations of descriptive scales can be significantly reduced without significant loss of model quality by removing insignificant gradations from the model. In this case, the dimensionality of the model will be significantly reduced, and its performance will increase accordingly.

Fig 14. Pareto curve for the significance of descriptive scales.

Fig 14

The Y-axis shows the total significance of the gradations of descriptive scales in %. The X-axis shows the gradations of descriptive scales in descending order of significance (as a percentage of their number).

3.3.  Results of SWOT-analysis

One well-known and accepted technique for strategic planning is SWOT (i.e., Strengths, Weaknesses, Opportunities, and Threats) analysis. This does not stop criticism, though, which is frequently valid, well-reasoned, and reasonable. The SWOT diagram for the Active Outcome class represents a feature contribution analysis specific to this class. In this context, SWOT is interpreted as an analytical framework highlighting the strengths, weaknesses, opportunities, and threats associated with feature–class relationships. The diagram illustrates the most significant associations derived from ASC analysis, where positive and negative contributions are encoded by color (red/blue), and the thickness of the connecting lines reflects the strength of each feature’s influence on the outcome. A thorough evaluation of SWOT analysis revealed numerous flaws, the main one being the requirement to consult specialists in order to determine the direction and magnitude of each factor’s influence [39]. Because of their expertise and professional experience, specialists do this instinctively. However, specialists’ talents are limited, and for a variety of reasons, they frequently are unable or unwilling to do this. Consequently, the challenge of performing a SWOT analysis without consulting professionals emerges [40,41]. By automating expert functions, such as determining the direction and magnitude of a factor’s influence based on empirical data, the Aidos system solves this issue [42]. The SWOT diagram of the Parasite class is shown in Fig 15.

Fig 15. SWOT diagram of the Parasite class.

Fig 15

The SWOT diagrams display the 12 most significant relationships, with the connection sign displayed in color (red plus, blue minus), and the value displayed by the thickness of the line. It is possible to display charts with only positive or only negative relationships. As a method for automated quantitative SWOT analysis, quantitative SWOT analysis of classes allows for the creation of a SWOT matrix for a particular class, indicating the degree of effect of facilitating and hindering elements directly based on the generated database. Factors that have a favorable impact on the control object’s transition to a certain state are listed in the class’s left column, followed by those that have no apparent impact and, finally, those that hinder the transition. It is feasible to identify the color spectrum range that will enable malaria to be reliably identified based on the findings of the SWOT analysis.

3.4. Limitations

Blood smear images must not exceed 800 x 600 pixels. Image digitization is performed using 12 spectral ranges. The Aidos system stores the intermediate data it can process in up to 2 GB of memory, limiting the number of images it can process. To ensure high reliability, blood smears should be of uniform size and shape (e.g., round) and use consistent staining and color hues.

Processing color images of blood smears is computationally intensive and demands substantial memory, as each fundus image contains hundreds of thousands of pixels. If large collections of smears are pre-digitized and stored, future spectral analysis needs to be applied only to new patient images, thereby shortening the malaria diagnostic process. To import converted data into the Aidos intelligent system, users must employ its universal software interface and specify the correct data format. All input files must be in Excel format. Although the current Aidos platform can handle millions of individual objects, its knowledge bases are limited to no more than 4,000 classes and 4,000 factor gradations.

Another significant limitation is the complexity of the Aidos user interface, which provides 55 operating modes (excluding the Exit mode). Even minor errors in data preparation or interface navigation can halt processing or generate inaccurate results. To reduce the learning curve, it is strongly recommended to review the training presentations available at [https://www.patreon.com/user?u=87599532], which explain the principles of automated system-cognitive analysis for solving complex diagnostic tasks [43].

4. Conclusion

A training database of blood smear parameters was successfully created. To evaluate its medical applicability, 18 test images were digitized—nine from malaria-infected patients and nine from healthy individuals. The digitization process for this test set of images took three minutes. The digitized data was saved to an Excel file. To digitize the test set of blood smear images, they were added in pairs to the training database, which required additional processing time. For large-scale malaria screening, new blood smear images should be added to the training database one by one, otherwise diagnostic reliability may be reduced. An expanded study of 436 digitized blood smears demonstrated 100% diagnostic accuracy, with all infected and healthy samples correctly classified. The number of blood smear images in the training set can be increased, but they must be digitized only once. Blood smear images in the test set can be digitized separately. This requires an additional step of merging the digitized data files from the training and test sets into a single Excel file. The additional one-time cost of digitizing a large number of training set images will only slightly increase the overall diagnostic time for test blood smear images.

Professor Lutsenko’s L1 measure of reliability of the INF3 model is 0.974. S-precision model INF3 equals 1.000, S-completeness equals 0.950. The confidence interval for malaria classification is defined by similarity values ranging from >25–100 when comparing digital smears with reference classes (parasites and healthy individuals). Images with a similarity score below 25 should be excluded from consideration as unrecognized. No such images were found in this study.

The Aidos intelligent system offers the following advantages over traditional microscopy:

  • The system automatically detects red blood cells and parasites;

  • Reduces the need for experienced microscopists;

  • Eliminates laboratory technician bias;

  • High diagnostic accuracy of blood smears, approximately 97–98%, is achieved, comparable to that of experienced parasitologists;

  • Analysis is completed in minutes, whereas manual microscopy can take 20–30 minutes per sample;

  • The system can automatically screen millions of cells, which is virtually impossible to do manually.

The Aidos intelligent system transforms malaria diagnosis from an expert manual procedure into an automated digital process. This is especially important for countries in Africa and Southeast Asia, which are remote from laboratories, and where experienced microscopists are often unavailable in endemic regions.

Supporting information

S1 File. Inp_data.

(XLSX)

pone.0348280.s001.xlsx (46.2KB, xlsx)

Acknowledgments

We thank Learning Engineer Nagadia from AURO University, Surat, Gujarat, India for providing the blood smear image samples.

Data Availability

All relevant data are within the paper and its Supporting Information files.

Funding Statement

The authors did not receive any funding for this research.

References

  • 1.Maqsood A, Farid MS, Khan MH, Grzegorzek M. Deep malaria parasite detection in thin blood smear microscopic images. Applied Sciences. 2021;11(5):2284. doi: 10.3390/app11052284 [DOI] [Google Scholar]
  • 2.World Health Organization. Malaria microscopy quality assurance manual-version 2. World Health Organization. 2016. [Google Scholar]
  • 3.Uzun Ozsahin D, Mustapha MT, Bartholomew Duwa B, Ozsahin I. Evaluating the Performance of Deep Learning Frameworks for Malaria Parasite Detection Using Microscopic Images of Peripheral Blood Smears. Diagnostics (Basel). 2022;12(11):2702. doi: 10.3390/diagnostics12112702 [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 4.Linder N, Turkki R, Walliander M, Mårtensson A, Diwan V, Rahtu E, et al. A malaria diagnostic tool based on computer vision screening and visualization of Plasmodium falciparum candidate areas in digitized blood smears. PLoS One. 2014;9(8):e104855. doi: 10.1371/journal.pone.0104855 [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 5.Maity M, Maity AK, Dutta PK, Chakraborty C. A web-accessible framework for automated storage with compression and textural classification of malaria parasite images. Int J Comput Appl. 2012;52:31–9. [Google Scholar]
  • 6.Shambhu S, Koundal D, Das P, Hoang VT, Tran-Trung K, Turabieh H. Computational Methods for Automated Analysis of Malaria Parasite Using Blood Smear Images: Recent Advances. Comput Intell Neurosci. 2022;2022:3626726. doi: 10.1155/2022/3626726 [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 7.Moon S, Lee S, Kim H, Freitas-Junior LH, Kang M, Ayong L, et al. An Image Analysis Algorithm for Malaria Parasite Stage Classification and Viability Quantification. PLoS ONE. 2013;8:e61812. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 8.Aregawi S, Li L, Miraglia CM. Malaria rapid diagnostic test and Giemsa – stained peripheral blood smear discrepancies in the diagnosis of Plasmodium ovale infection in New England. Clin Lab Sci. 2017;30(2):75–83. doi: 10.29074/ascls.30.2.75 [DOI] [Google Scholar]
  • 9.Maltha J, Gillet P, Jacobs J. Malaria rapid diagnostic tests in travel medicine. Clin Microbiol Infect. 2013;19:408–15. [DOI] [PubMed] [Google Scholar]
  • 10.de Almeida JG, Gudgin E, Besser M, Dunn WG, Cooper J, Haferlach T, et al. Computational analysis of peripheral blood smears detects disease-associated cytomorphologies. Nat Commun. 2023;14(1):4378. doi: 10.1038/s41467-023-39676-y [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 11.Tangpukdee N, Duangdee C, Wilairatana P, Krudsood S. Malaria Diagnosis: A Brief Overview. Korean J Parasitol. 2009;47(2):93–102. doi: 10.3347/kjp.2009.47.2.93 [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 12.Rajaraman S, Antani SK, Poostchi M, Silamut K, Hossain MA, Maude RJ, et al. Pre-trained convolutional neural networks as feature extractors toward improved malaria parasite detection in thin blood smear images. PeerJ. 2018;6:e4568. doi: 10.7717/peerj.4568 [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 13.Yang F, Yu H, Silamut K, Maude RJ, Jaeger S, Antani S. Smartphone-Supported Malaria Diagnosis Based on Deep Learning. Lecture Notes in Computer Science. Springer International Publishing. 2019. 73–80. doi: 10.1007/978-3-030-32692-0_9 [DOI] [Google Scholar]
  • 14.Rosadoa L, Correia da Costa JM, Elias D, Cardoso JS. Automated detection of malaria parasites on thick blood smears via mobile devices. Procedia Computer Science. 2016;90:138–44. [Google Scholar]
  • 15.Masud M, Alhumyani H, Alshamrani SS, Cheikhrouhou O, Ibrahim S, Muhammad G, et al. Leveraging Deep Learning Techniques for Malaria Parasite Detection Using Mobile Application. Wireless Communications and Mobile Computing. 2020;2020:1–15. doi: 10.1155/2020/8895429 [DOI] [Google Scholar]
  • 16.Dong Y, Jiang Z, Shen H, David Pan W, Williams LA, Reddy VVB, et al. Evaluations of deep convolutional neural networks for automatic identification of malaria infected cells. In: 2017 IEEE EMBS International Conference on Biomedical & Health Informatics (BHI), 2017. 101–4. doi: 10.1109/bhi.2017.7897215 [DOI] [Google Scholar]
  • 17.Bibin D, Nair MS, Punitha P. Malaria Parasite Detection From Peripheral Blood Smear Images Using Deep Belief Networks. IEEE Access. 2017;5:9099–108. doi: 10.1109/access.2017.2705642 [DOI] [Google Scholar]
  • 18.Kumar S, Vardhan H, Priya S, Kumar A. Malaria detection using deep convolution neural network. IJAER. 2020;4(3):6–12. [Google Scholar]
  • 19.Magotra V, Rohil MK. Malaria Diagnosis Using a Lightweight Deep Convolutional Neural Network. Int J Telemed Appl. 2022;2022:4176982. doi: 10.1155/2022/4176982 [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 20.Madhu G, Mohamed AW, Kautish S, Shah MA, Ali I. Intelligent diagnostic model for malaria parasite detection and classification using imperative inception-based capsule neural networks. Sci Rep. 2023;13(1):13377. doi: 10.1038/s41598-023-40317-z [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 21.Beck H-P. Digital microscopy and artificial intelligence could profoundly contribute to malaria diagnosis in elimination settings. Front Artif Intell. 2022;5:510483. doi: 10.3389/frai.2022.510483 [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 22.Fatima T, Farid MS. Automatic detection of Plasmodium parasites from microscopic blood images. J Parasit Dis. 2020;44(1):69–78. doi: 10.1007/s12639-019-01163-x [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 23.Poostchi M, Silamut K, Maude RJ, Jaeger S, Thoma G. Image analysis and machine learning for detecting malaria. Transl Res. 2018;194:36–55. doi: 10.1016/j.trsl.2017.12.004 [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 24.Han JH, Han ET. Malaria diagnostics: from traditional techniques to cutting-edge solutions. Ann Clin Microbiol. 2024;27(3). doi: 10.5145/ACM.2024.27.3.2 [DOI] [Google Scholar]
  • 25.Shewajo FA, Fante KA. Tile-based microscopic image processing for malaria screening using a deep learning approach. BMC Med Imaging. 2023;23(1):39. doi: 10.1186/s12880-023-00993-9 [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 26.Lutsenko EV. Automated System-Cognitive Analysis of the Dependence of Agrophysical Indicators of the Soil on its Processing, Fertilizers and the Phase of Wheat Vegetation. 2022. doi: 10.13140/RG.2.2.34682.77766 [DOI] [Google Scholar]
  • 27.Lutsenko EV. ASC-Analysis and the Aidos System as a Method and Tools for Solving Problems. 2021. doi: 10.13140/RG.2.2.29823.74407 [DOI] [Google Scholar]
  • 28.Lutsenko EV, Bandyk DK, Troshin LP. The solution of problems of ampelography by using ASC-analysis of images of leaves in their external contours (generalization, abstraction, classification and identification). Scientific Journal of KubGAU. 2015;112(08):1–49. [Google Scholar]
  • 29.Ichhpujani P, Ryabtsev V, Utkina T. Intellectual System Diagnostics Glaucoma. Jurnal Ilmiah Teknik Elektro Komputer dan Informatika (JITEKI). 2023;9(4):1045–59. doi: 10.26555/jiteki.v9i4.26969 [DOI] [Google Scholar]
  • 30.Lutsenko EV. System-cognitive analysis of images (generalization, abstraction, classification and identification). Scientific journal of KubSAU. 2009;(02 (046)):146–64. [Google Scholar]
  • 31.Malaria Dataset. https://www.kaggle.com/datasets/meetnagadia/malaria-dataset
  • 32.Lutsenko EV. Theoretical Foundations, Mathematical Model and Software Tools for Automated System-Cognitive Analysis. 2020. [Google Scholar]
  • 33.Lutsenko EV, Bandyk DK. Interface for entering images into the Aidos system (Aidos-img subsystem). 2015. http://lc.kubagro.ru/aidos/2015618040.jpg [Google Scholar]
  • 34.Lutsenko EV. Scenario and spectral automated system-cognitive analysis. 2021. doi: 10.13140/RG.2.2.22981.37608 [DOI] [Google Scholar]
  • 35.Al-Ansi AMA, Ryabtsev V, Utkina T. The Use of Large Databases for Diagnosing Human Diseases at Early Stage. IJAIDM. 2023;6(2):197. doi: 10.24014/ijaidm.v6i2.24525 [DOI] [Google Scholar]
  • 36.Al-Ansi AM, Almadi M, Ichhpujani P, Ryabtsev V. Aidos System Prediction of Myopia in Children in Early Education Stages. Jurnal Ilmiah Teknik Elektro Komputer dan Informatika (JITEKI). 2023;9(2):411–9. doi: 10.26555/jiteki.v9i2.26292 [DOI] [Google Scholar]
  • 37.Lutsenko EV. Identification of atypical objects and artifacts in the source data, assignment to new classes of atypical objects and removal of artifacts in mathematical models of automated system-cognitive analysis. Scientific Journal of KubGAU. 2022;184(10):1–50. doi: 10.13140/RG.2.2.22198.98881 [DOI] [Google Scholar]
  • 38.Orlov AI, Lutsenko EV, Loiko VI. Organizational, economic, mathematical and software support for controlling, innovation and management: monograph. Falko SG. Krasnodar: KubSAU. 2016. [Google Scholar]
  • 39.Orlov AI, Lutsenko EV. Systemic fuzzy interval mathematics. Krasnodar: KubSAU. 2014. [Google Scholar]
  • 40.King T, Freyn S, Morrison J. SWOT analysis problems and solutions: Practitioners’ feedback into the ongoing academic debate. JISIB. 2023;13(1):30–42. doi: 10.37380/jisib.v13i1.989 [DOI] [Google Scholar]
  • 41.Palazzo M. The SWOT analysis: an evolving decision-making model. 2024. [Google Scholar]
  • 42.Lutsenko EV. Methods of artificial intelligence: Textbook. Manual. Krasnodar: VTsSKI “Aidos”. 2020. [Google Scholar]
  • 43.Ryabtsev VG. Lectures on Electronic Medicine. 2023. https://www.patreon.com/user?u=87599532 [Google Scholar]

Decision Letter 0

Segun OYEDEJI

8 Sep 2025

-->PONE-D-25-34548-->-->Intelligent Automation Cognitive System for Malaria Diagnosis Using Digital Blood Smears-->-->PLOS ONE

Dear Dr. Saleh,

Thank you for submitting your manuscript to PLOS ONE. After careful consideration, we feel that the study addresses an important health topic, but does not fully meet PLOS ONE’s publication criteria as it currently stands. Therefore, we invite you to submit a revised version of the manuscript that addresses the points raised during the review process.

==============================

First, there are substantial grammatical issues majorly bordering on the use of non-professional terminologies. I will suggest that the authors engage a Professional (biologist, parasitologist, epidemiologist or clinician) with sufficient kowledge of the disease to assist with the write-up.

Second, it was observed that standard performance metrics such as sensitivity, specificity, accuracy, precision, recall, F1 score, and ROC/AUC score, were not used to evaluate the result of the Model. The manuscript would improve significantly with the inclusion of the above standard performance metrics.

Third, evaluating only 18 images for the possibility of using the spectral analysis method for medical purposes is grossly inadequate. The authors could consider significant increase in this number to provide sufficient evidence to validate the results. The authors should also provide the procedure for this in the Methods Section.

Please go through each of the Reviewer’s Comments to address each point raised.

==============================

Please submit your revised manuscript by Oct 23 2025 11:59PM. If you will need more time than this to complete your revisions, please reply to this message or contact the journal office at plosone@plos.org. When you're ready to submit your revision, log on to https://www.editorialmanager.com/pone/ and select the ’submissions Needing Revision' folder to locate your manuscript file.

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If you would like to make changes to your financial disclosure, please include your updated statement in your cover letter. Guidelines for resubmitting your figure files are available below the reviewer comments at the end of this letter.

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We look forward to receiving your revised manuscript.

Kind regards,

Segun Isaac OYEDEJI, Ph.D

Academic Editor

PLOS ONE

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When you resubmit, please ensure that you provide the correct grant numbers for the awards you received for your study in the ‘Funding Information’ section.

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King Salman Center for Disability Research     KSRG-2024-281.

Please state what role the funders took in the study. If the funders had no role, please state: "The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript."

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7. Please amend your authorship list in your manuscript file to include author Marwa Saleh.

8. Please amend the manuscript submission data (via Edit Submission) to include author Marwa A. Saleh.

9. If the reviewer comments include a recommendation to cite specific previously published works, please review and evaluate these publications to determine whether they are relevant and should be cited. There is no requirement to cite these works unless the editor has indicated otherwise.

[Note: HTML markup is below. Please do not edit.]

Reviewers' comments:

Reviewer’s Responses to Questions

-->Comments to the Author

1. Is the manuscript technically sound, and do the data support the conclusions?

The manuscript must describe a technically sound piece of scientific research with data that supports the conclusions. Experiments must have been conducted rigorously, with appropriate controls, replication, and sample sizes. The conclusions must be drawn appropriately based on the data presented. -->

Reviewer #1: Yes

Reviewer #2: Partly

Reviewer #3: No

**********

-->2. Has the statistical analysis been performed appropriately and rigorously? -->

Reviewer #1: Yes

Reviewer #2: I Don't Know

Reviewer #3: No

**********

-->3. Have the authors made all data underlying the findings in their manuscript fully available?

The PLOS Data policy requires authors to make all data underlying the findings described in their manuscript fully available without restriction, with rare exception (please refer to the Data Availability Statement in the manuscript PDF file). The data should be provided as part of the manuscript or its supporting information, or deposited to a public repository. For example, in addition to summary statistics, the data points behind means, medians and variance measures should be available. If there are restrictions on publicly sharing data—e.g. participant privacy or use of data from a third party—those must be specified.-->

Reviewer #1: Yes

Reviewer #2: Yes

Reviewer #3: No

**********

-->4. Is the manuscript presented in an intelligible fashion and written in standard English?

PLOS ONE does not copyedit accepted manuscripts, so the language in submitted articles must be clear, correct, and unambiguous. Any typographical or grammatical errors should be corrected at revision, so please note any specific errors here.-->

Reviewer #1: Yes

Reviewer #2: No

Reviewer #3: No

**********

-->5. Review Comments to the Author

Please use the space provided to explain your answers to the questions above. You may also include additional comments for the author, including concerns about dual publication, research ethics, or publication ethics. (Please upload your review as an attachment if it exceeds 20,000 characters)-->

Reviewer #1: Compulsory revisions

Abstract

The graphical abstract needs proper labelling so that readers can quickly understand the process, key components, or findings.

Introduction

• The introduction part contains too much technical detail, and dense information such as laser energy in nJ, types of filters without context. It is better to write in a clear form for better interation with the reader.

• The WHO death statistics quoted is old, use a recent report.

• There is inconsistent flow in the write up: the transition from classical methods to deep learning is abrupt. Then the Eidos system is introduced without first motivating the gap that it fills. Give a brief introduction of Eidos and ASC.

Methods

• Figure 1 and 2 looks blurred, replace with clearer images

Results and Discussion

• You mentioned that using the INF3 model, digitalized blood smear image data were recognized with a high degree of similarity.

Give a clear explanation of this model before reporting it in this section.

• You mention using SWOT analysis, can you give the full meaning and a brief description of the analysis.

• Figure 13 needs to be properly labelled at the X and Y axis.

Reviewer #2: Dear Editor,

Please find below, my comments and suggestions on the reviewed manuscript:

General:

The manuscript describes a study which employed an automated system-cognitive analysis of digitized blood smears as a diagnostic tool for malaria. This is a step forward in malaria diagnosis. However, there is the need for a critical revision of the manuscript to attend to the reporting style in order to ensure the validity and reliability of the findings. There is also a need for clarity and consistency throughout the manuscript so as to strengthen its overall quality and contribution to the understanding of the use of automation cognitive system for digital blood smears in malaria diagnostics.

• The English language quality of the manuscript needs improvement to make for better reading.

• The technical language of the manuscript needs to be revised to make it more scientifically sound, and easily comprehensible across the readership community.

• The authors need to state/include at least, the genus of the causative agent(s) of malaria (i.e., Plasmodium) in the manuscript. This can be used interchangeably with malaria where applicable.

Graphical Abstract:

• Line 1: The authors should consider including the highlighted words: “…. by species of Plasmodium parasites …”

• Line 2: The genus name, Anopheles, should be italicized; and subsequently, throughout the manuscript.

Introduction:

• Line 1: “contract” is a better word for ‘’obtain’’. Also consider including the highlighted texts: “…. female Anopheles mosquitoes carrying …..”

• Lines 3-5: The statement “According to the ……., 438,000 … in 2015, … 620,000 … in 2017, …. 300-500 million …… [2]” is obsolete and should be replaced with more recent data. The authors are advised to refer to: (263 million and 597,000 malaria cases and deaths respectively, in 2023 [World Malaria Report, 2024]).

• Line 12: The authors need to be more specific with the statement “To count the number of infected cells in a single blood film ….” by indicating the type of cells and blood film (thin or thick)?

• Lines 16-19: The statement “Rapid tests …….. impact patient treatment [7].” should be recast for better comprehension. For instance, “Rapid tests” could be re-written as “rapid diagnostic tests (RDTs)”

• Third paragraph: The authors need to be more specific with the statement: ”Sophisticated image processing ….. automate malaria detection [8]“. ….. using shape, color, intensity, size and texture …..“. The question is: Of what? The malaria parasite, or blood cells?

• Third paragraph: The authors need to be more specific with the statement: ”Lasers …. the cells of interest from blood smears [8]“. The authors should state the cells of interest.

• Third paragraph: Consider revising the statement as highlighted: ”Images of diseased (infected) and uninfected red blood cells (RBCs) … [16, 17]“.

• Third paragraph: Consider revising the statement as highlighted: ” … films that can identify low parasitemia [21, 22]“.

• Third paragraph. If what the authors mean by tiny in the phrase? … with identifying tiny malaria parasites in blood … [24]“ is in regard to the malaria parasite density, then they should consider using low.

• Page 4: Line 2: “….. and solving several other problems.” should be referenced.

• Last paragraph. Consider the following “The present work aims to ….”.

Methods

2.1. Formalization of the subject Area:

AURO University is in India. However, it is not in Mumbai, Maharashtra, as stated by the authors, but rather in Surat, Gujarat.

• Consider revising “…. 550 images of infected and uninfected blood smears ….” as: “…. 550 images of malaria (Plasmodium)-positive and negative blood smears, ….”

• ….. can be used for training and testing. Respectively???

• “Examples of malaria-unaffected ….” to “Examples of malaria-negative ….”

• “… examples of affected …. are demonstrated ….” to “… examples of malaria-positive …. are shown ….”

• Consider revising Titles of Figs. 1 and 2 as Malaria/Plasmodium-negative blood smear samples and Malaria/Plasmodium-positive blood smear samples, respectively.

• Page 5, lines 7/8: “The spectra ….. are then compared with those of classes”. What classes? According to the Eidos system or what?

• Last paragraph: “…. starting with column Dare …” to “…. starting with column D are …”

2.2. Synthesis of statistical and system-cognitive models:

• Consider revising Title of Fig. 6 as Matrix of conditional percentage distributions.

• Page 8: Line 9: What is the full meaning of the acronym INF3?

• Page 9: Line 5: Consider replacing ‘sick’ with ‘infected’ and ‘healthy’ with ‘uninfected’.

Results and Discussion

3.1. Results of recognition of the examined blood smears:

• Fig. 9: Given that a malaria recognition accuracy rate of 100% was recorded with several of the studied smears, the authors should consider showing at least, one of such in Fig. 9.

• “All 100% of infected ……” Better to use either of the two (i.e., All or 100%).

• Consider changing “The circular histogram of malaria ….” to “The pie chart of malaria …...”

• Fig. 10: This is not a circular histogram, but rather, a pie chart! Also, the , (commas) in the listed values should be replaced by . (i.e., dots).

• Page 9, Last paragraph: Consider replacing “The right column ……. colors” with “Legend:”. This should be placed atop the column showing the different colors and values.

• Fig. 11: The data shown here is a duplication of Fig. 9! Moreover, Fig. 11 is supposed to show the results of recognition of digitized blood smears of healthy patients!

3.2. Results of the selection of light spectra for the study:

• Consider revising “The Eidos system allows you to determine …..” as “The Eidos system allows for the determination of …” and “The effect of light spectra is …..” as “The result of this is …….”

• Page 11, Line 1: The full meaning of SWOT (i.e., Strengths, weaknesses, Opportunities, and threats) should be stated in parenthesis.

Conclusion

• Line 6: i.e., infected blood masks?

Reviewer #3: The manuscript proposes using the “Eidos” intelligent system to diagnose malaria from digitized thin blood smear images by converting images into “12 light spectra” and classifying them via an automated system-cognitive (ASC) approach. The authors report an “average similarity value” of 66.965% for infected samples, “no false positives” for healthy samples, and a processing time of ~10 seconds for 18 new images.

While rapid, low-cost diagnostic support is an important goal, the current study has substantial methodological, reporting, and reproducibility limitations that prevent a reliable assessment of performance or utility- see attached document please.

**********

-->6. PLOS authors have the option to publish the peer review history of their article (what does this mean?). If published, this will include your full peer review and any attached files.

If you choose “no”, your identity will remain anonymous but your review may still be made public.

Do you want your identity to be public for this peer review?   For information about this choice, including consent withdrawal, please see our Privacy Policy.-->

Reviewer #1: Yes:   Adedolapo Blessing Olorunfemi

Reviewer #2: No

Reviewer #3: No

**********

[NOTE: If reviewer comments were submitted as an attachment file, they will be attached to this email and accessible via the submission site. Please log into your account, locate the manuscript record, and check for the action link "View Attachments". If this link does not appear, there are no attachment files.]

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Attachment

Submitted filename: Reviewers Comments.pdf

pone.0348280.s002.pdf (318.9KB, pdf)
Attachment

Submitted filename: Reviewers report.docx

pone.0348280.s003.docx (17.7KB, docx)
PLoS One. 2026 Jun 18;21(6):e0348280. doi: 10.1371/journal.pone.0348280.r002

Author response to Decision Letter 1


4 Oct 2025

all responses for respected reviewers has been attached as file in submission and all changes have been included in manuscript file revised

Attachment

Submitted filename: Response to Reviewers.docx

pone.0348280.s004.docx (25.3KB, docx)

Decision Letter 1

Segun OYEDEJI, José Luiz Vieira

18 Feb 2026

-->PONE-D-25-34548R1-->-->Intelligent Automation Cognitive System for Malaria Diagnosis Using Digital Blood Smears-->-->PLOS One

Dear Dr. A.Saleh,-->-->

Thank you for submitting your manuscript to PLOS ONE. After careful consideration, we feel that it has merit but does not fully meet PLOS ONE’s publication criteria as it currently stands. Therefore, we invite you to submit a revised version of the manuscript that addresses the points raised during the review process.

ACADEMIC EDITOR:   -->-->Dear Dr. A.Saleh,-->--> -->-->Please carefully review the comments provided by Reviewer Olaiya Folorunsho and incorporate the necessary revisions into the manuscript, particularly those concerning validation and reproducibility, as well as the discussion of limitations and practical implementation. This decision is  justified on PLOS ONE’s publication criteria and not, for example, on novelty or perceived impact.

Please submit your revised manuscript by Apr 04 2026 11:59PM. If you will need more time than this to complete your revisions, please reply to this message or contact the journal office at plosone@plos.org. When you're ready to submit your revision, log on to https://www.editorialmanager.com/pone/ and select the ’submissions Needing Revision' folder to locate your manuscript file.

Please include the following items when submitting your revised manuscript:-->

  • A letter that responds to each point raised by the academic editor and reviewer(s). You should upload this letter as a separate file labeled 'Response to Reviewers'.

  • A marked-up copy of your manuscript that highlights changes made to the original version. You should upload this as a separate file labeled 'Revised Manuscript with Track Changes'.

  • An unmarked version of your revised paper without tracked changes. You should upload this as a separate file labeled 'Manuscript'.

-->If you would like to make changes to your financial disclosure, please include your updated statement in your cover letter. Guidelines for resubmitting your figure files are available below the reviewer comments at the end of this letter.

If applicable, we recommend that you deposit your laboratory protocols in protocols.io to enhance the reproducibility of your results. Protocols.io assigns your protocol its own identifier (DOI) so that it can be cited independently in the future. For instructions see: https://journals.plos.org/plosone/s/submission-guidelines#loc-laboratory-protocols. Additionally, PLOS ONE offers an option for publishing peer-reviewed Lab Protocol articles, which describe protocols hosted on protocols.io. Read more information on sharing protocols at https://plos.org/protocols?utm_medium=editorial-email&utm_source=authorletters&utm_campaign=protocols.

We look forward to receiving your revised manuscript.

Kind regards,

José Luiz Fernandes Vieira

Academic Editor

PLOS One

Journal Requirements:

If the reviewer comments include a recommendation to cite specific previously published works, please review and evaluate these publications to determine whether they are relevant and should be cited. There is no requirement to cite these works unless the editor has indicated otherwise.

Please review your reference list to ensure that it is complete and correct. If you have cited papers that have been retracted, please include the rationale for doing so in the manuscript text, or remove these references and replace them with relevant current references. Any changes to the reference list should be mentioned in the rebuttal letter that accompanies your revised manuscript. If you need to cite a retracted article, indicate the article’s retracted status in the References list and also include a citation and full reference for the retraction notice.

[Note: HTML markup is below. Please do not edit.]

Reviewers' comments:

Reviewer’s Responses to Questions

-->Comments to the Author

1. If the authors have adequately addressed your comments raised in a previous round of review and you feel that this manuscript is now acceptable for publication, you may indicate that here to bypass the “Comments to the Author” section, enter your conflict of interest statement in the “Confidential to Editor” section, and submit your "Accept" recommendation.-->

Reviewer #1: All comments have been addressed

Reviewer #4: (No Response)

**********

-->2. Is the manuscript technically sound, and do the data support the conclusions?

The manuscript must describe a technically sound piece of scientific research with data that supports the conclusions. Experiments must have been conducted rigorously, with appropriate controls, replication, and sample sizes. The conclusions must be drawn appropriately based on the data presented. -->

Reviewer #1: Yes

Reviewer #4: (No Response)

**********

-->3. Has the statistical analysis been performed appropriately and rigorously? -->

Reviewer #1: Yes

Reviewer #4: (No Response)

**********

-->4. Have the authors made all data underlying the findings in their manuscript fully available?

The PLOS Data policy requires authors to make all data underlying the findings described in their manuscript fully available without restriction, with rare exception (please refer to the Data Availability Statement in the manuscript PDF file). The data should be provided as part of the manuscript or its supporting information, or deposited to a public repository. For example, in addition to summary statistics, the data points behind means, medians and variance measures should be available. If there are restrictions on publicly sharing data—e.g. participant privacy or use of data from a third party—those must be specified.-->

Reviewer #1: Yes

Reviewer #4: (No Response)

**********

-->5. Is the manuscript presented in an intelligible fashion and written in standard English?

PLOS ONE does not copyedit accepted manuscripts, so the language in submitted articles must be clear, correct, and unambiguous. Any typographical or grammatical errors should be corrected at revision, so please note any specific errors here.-->

Reviewer #1: Yes

Reviewer #4: Yes

**********

-->6. Review Comments to the Author

Please use the space provided to explain your answers to the questions above. You may also include additional comments for the author, including concerns about dual publication, research ethics, or publication ethics. (Please upload your review as an attachment if it exceeds 20,000 characters)-->

Reviewer #1: Compulsory revisions

Abstract

The graphical abstract needs proper labelling so that readers can quickly understand the process, key components, or findings.

Comment have been addressed

Introduction

• The introduction part contains too much technical detail, and dense information such as laser energy in nJ, types of filters without context. It is better to write in a clear form for better interation with the reader.

Comment have been addressed

• The WHO death statistics quoted is old, use a recent report.

Comment have been addressed

• There is inconsistent flow in the write up: the transition from classical methods to deep learning is abrupt. Then the Eidos system is introduced without first motivating the gap that it fills. Give a brief introduction of Eidos and ASC.

Comment have been addressed

Methods

• Figure 1 and 2 looks blurred, replace with clearer images

Comment have been addressed

Results and Discussion

• You mentioned that using the INF3 model, digitalized blood smear image data were recognized with a high degree of similarity.

Give a clear explanation of this model before reporting it in this section.

Comment have been addressed

• You mention using SWOT analysis, can you give the full meaning and a brief description of the analysis.

Comment have been addressed

• Figure 13 needs to be properly labelled at the X and Y axis.

Comment have been addressed

Reviewer #4: a) Explicit mention of ethical approvals or data usage permissions for patient samples should be included, even if the dataset is public.

b) Potential clinical implications and how the automated system would support healthcare decision-making should be discussed.

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Reviewer #1: Yes:   Adedolapo Blessing Olorunfemi

Reviewer #4: Yes:   Olaiya Folorunsho

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Attachment

Submitted filename: REVIEWER REPORT.docx

pone.0348280.s005.docx (16.6KB, docx)
PLoS One. 2026 Jun 18;21(6):e0348280. doi: 10.1371/journal.pone.0348280.r004

Author response to Decision Letter 2


10 Apr 2026

Response to Reviewers

Manuscript Title: Intelligent Automation Cognitive System for Malaria Diagnosis Using Digital Blood Smears

Journal: PLOS ONE

Dear Academic Editor and Reviewers,

We sincerely thank you for your careful evaluation of our manuscript and for the constructive comments. We appreciate the opportunity to revise our work. We have carefully considered all comments and revised the manuscript accordingly. All changes have been incorporated into the revised manuscript and highlighted in the tracked-changes version.

Below, we provide a point-by-point response.

Response to the Academic Editor

Fund statement

The funding has been removed, as the King Salman Center for Disability Research does not currently support this research due to exceeding the contractual deadline for completing the publication process.

Comment:

Please carefully review the comments provided by Reviewer Olaiya Folorunsho and incorporate the necessary revisions into the manuscript, particularly those concerning validation and reproducibility, as well as the discussion of limitations and practical implementation.

Response:

Thank you for this guidance. We revised the manuscript to strengthen the presentation of validation, reproducibility, limitations, and practical implementation. Specifically:

1. We clarified the dataset source and data usage conditions, including the public availability of the malaria blood smear images used in the study.

2. We explicitly stated that the study used a publicly available, de-identified dataset and that no institutional review board approval or informed consent was required.

3. We expanded the description of the digitization and analysis workflow, including the role of the Aidos system, the use of 12 spectral ranges, and the practical steps required for importing and processing images.

4. We added a clearer discussion of reproducibility-related issues, including file format requirements, image size constraints, computational limitations, and the need for consistent smear quality and staining.

5. We expanded the discussion of practical implementation by explaining how the system can support preliminary malaria screening and assist healthcare workers, especially in settings with limited access to expert microscopists.

6. We revised the limitations section to better reflect technical and operational constraints.

These revisions were incorporated into the Methods, Discussion, Limitations, Ethics statement, Source Data, and Conclusion sections.

Response to Reviewer 1

We thank Reviewer-1 for the positive assessment and for confirming that the previous comments have been addressed.

General comment

The graphical abstract needs proper labelling so that readers can quickly understand the process, key components, or findings.

Comment have been addressed

• The introduction part contains too much technical detail, and dense information such as laser energy in nJ, types of filters without context. It is better to write in a clear form for better interation with the reader.

Comment have been addressed

• The WHO death statistics quoted is old, use a recent report.

Comment have been addressed

• There is inconsistent flow in the write up: the transition from classical methods to deep learning is abrupt. Then the Eidos system is introduced without first motivating the gap that it fills. Give a brief introduction of Eidos and ASC.

Comment have been addressed

• Figure 1 and 2 looks blurred, replace with clearer images

Comment have been addressed

Results and Discussion

• You mentioned that using the INF3 model, digitalized blood smear image data were recognized with a high degree of similarity.

Give a clear explanation of this model before reporting it in this section.

Comment have been addressed

• You mention using SWOT analysis, can you give the full meaning and a brief description of the analysis.

Comment have been addressed

• Figure 13 needs to be properly labelled at the X and Y axis.

Response:

This comment has been addressed. Figure 14 has been updated with clearly defined axis labels to improve interpretability. Specifically, the X-axis now represents the gradations of descriptive scales arranged in descending order of significance (expressed as a percentage of their total number), while the Y-axis represents the cumulative significance of these gradations (expressed as a percentage).

We are grateful for your careful review and positive evaluation of the revised manuscript.

Response to Reviewer 4

We sincerely thank Reviewer-4 for the valuable comments, which helped us improve the manuscript.

Comment 4a

Comment:

Explicit mention of ethical approvals or data usage permissions for patient samples should be included, even if the dataset is public.

Response:

Thank you for this important comment. We have now explicitly clarified the ethical and data-use status of the dataset in the revised manuscript.

Specifically, we added that:

• The blood smear photographs were obtained from a publicly available Kaggle repository;

• The dataset is de-identified, and no patient names or personally identifiable information are provided;

• Therefore, ethical approval and informed consent were not required for the present study;

• Access to the dataset does not require approval from an ethics committee or any other institutional body.

These clarifications have been added in the methods section and further summarized in the ethics statement and third party material.

Added text in the manuscript:

“The blood smear photographs in the Kaggle repository dataset are publicly available, and the patient names are not provided, so no ethical approval or permission to use the samples for research is required.”

We also added in the Ethics statement section:

“This was not a human population study; therefore, approval by the institutional review board and informed consent were not required.”

Comment 4b

Comment:

Potential clinical implications and how the automated system would support healthcare decision-making should be discussed.

Response:

Thank you. We have expanded the manuscript to better explain the clinical relevance and practical role of the proposed system.

In the revised Discussion and Conclusion, we now clarify that the proposed automated system is intended as a preliminary diagnostic support tool, not as a replacement for clinical judgment. The system can support healthcare decision-making in several ways:

The Aidos intelligent system offers the following advantages over traditional microscopy:

The system automatically detects red blood cells and parasites;

Reduces the need for experienced microscopists;

Eliminates laboratory technician bias;

High diagnostic accuracy of blood smears, approximately 97–98%, is achieved, comparable to that of experienced parasitologists;

Analysis is completed in minutes, whereas manual microscopy can take 20–30 minutes per sample;

The system can automatically screen millions of cells, which is virtually impossible to do manually.

We also clarified that the system provides instant diagnostic support and may be especially valuable in Africa and Southeast Asia, where access to specialized laboratory expertise is often limited.

Added/strengthened text in the manuscript includes statements such as:

• “The automated system-cognitive analysis of digitized blood smears provides instant diagnostic support.”

• “It allows medical workers with limited knowledge in microscopy and artificial intelligence to perform diagnostics.”

• “The Aidos intelligent system transforms malaria diagnosis from an expert manual procedure into an automated digital process.”

• “This is especially important for countries in Africa and Southeast Asia, which are remote from laboratories, and where experienced microscopists are often unavailable in endemic regions.”

We also expanded the limitations section to make clear that this system should be viewed as a supportive tool and that practical implementation depends on image quality, correct digitization, and proper operation of the Aidos interface.

Additional Revisions Made in the Manuscript

In addition to the reviewer-specific comments above, we made several improvements for clarity and completeness:

• Clarified the description of the INF3 model, including its role as an information/knowledge model used for recognition and similarity assessment;

The description of the INF3 model has been refined, emphasizing its role as an information/knowledge-based model for recognition and similarity assessment. According to E. V. Lutsenko, the L1 reliability measure for the INF3 model is 0.974. The model demonstrates high performance, with S-precision equal to 1.000 and S-completeness equal to 0.950. Furthermore, the confidence interval for malaria classification is defined by similarity values ranging from >25 to 100 when comparing digital blood smears with reference classes (parasite-infected and healthy samples). Images with similarity scores below 25 are considered unrecognized and excluded from analysis. Notably, no such cases were observed in this study.

• Clarified the meaning of SWOT analysis as “Strengths, Weaknesses, Opportunities, and Threats” and briefly explained its use in the study;

The meaning of SWOT analysis (Strengths, Weaknesses, Opportunities, and Threats) has been explicitly clarified and contextualized within the study. The SWOT diagram for the Active Outcome class represents a feature contribution analysis specific to this class. It illustrates the most significant feature–class relationships derived from ASC analysis, where positive and negative contributions are encoded by color (red/blue), and the thickness of the connecting lines reflects the strength of influence.

• Updated and refined the limitations section;

The limitations section has been expanded and clarified. Specifically, blood smear images must not exceed a resolution of 800 × 600 pixels. Image digitization is conducted across 12 spectral ranges. The Aidos system can process intermediate data up to 2 GB of memory, which limits the number of images processed simultaneously. Additionally, to ensure high reliability, blood smear samples should be standardized in size and shape (e.g., circular) and prepared using consistent staining protocols and color characteristics.

• Ensured that the Data Availability, Ethics statement, and Source Data sections clearly describe the public dataset and its conditions of use;

The Data Availability, Ethics Statement, and Source Data sections have been revised to clearly describe the public dataset used and the associated conditions of use.

• Improved the language and organization of the manuscript to increase readability.

The manuscript has undergone thorough language editing and structural reorganization to improve clarity, coherence, and overall readability.

Closing Statement

We appreciate the reviewers’ and editor’s thoughtful comments, which have helped us improve the manuscript substantially. We hope that the revised version now meets the journal’s requirements and is suitable for publication in PLOS ONE.

Sincerely,

On behalf of all authors

Dr. A. Saleh

Attachment

Submitted filename: Responses to Reviewers.docx

pone.0348280.s006.docx (24.8KB, docx)

Decision Letter 2

Segun OYEDEJI, José Luiz Vieira, José Luiz Vieira

14 Apr 2026

Intelligent Automation Cognitive System for Malaria Diagnosis Using Digital Blood Smears

PONE-D-25-34548R2

Dear Dra. Saleh

We’re pleased to inform you that your manuscript has been judged scientifically suitable for publication and will be formally accepted for publication once it meets all outstanding technical requirements.

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Kind regards,

José Luiz Fernandes Vieira

Academic Editor

PLOS One

Additional Editor Comments

Following a thorough and rigorous evaluation process, we are pleased to note that all the points raised by the reviewers during the previous rounds of revision have been meticulously addressed in your final manuscript. We appreciate the dedication and effort you and your co-authors have put into refining the work, which has significantly contributed to its clarity and scientific robustness.

Acceptance letter

Segun OYEDEJI, José Luiz Vieira, José Luiz Vieira

PONE-D-25-34548R2

PLOS One

Dear Dr. A.Saleh,

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Academic Editor

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Associated Data

    This section collects any data citations, data availability statements, or supplementary materials included in this article.

    Supplementary Materials

    S1 File. Inp_data.

    (XLSX)

    pone.0348280.s001.xlsx (46.2KB, xlsx)
    Attachment

    Submitted filename: Reviewers Comments.pdf

    pone.0348280.s002.pdf (318.9KB, pdf)
    Attachment

    Submitted filename: Reviewers report.docx

    pone.0348280.s003.docx (17.7KB, docx)
    Attachment

    Submitted filename: Response to Reviewers.docx

    pone.0348280.s004.docx (25.3KB, docx)
    Attachment

    Submitted filename: REVIEWER REPORT.docx

    pone.0348280.s005.docx (16.6KB, docx)
    Attachment

    Submitted filename: Responses to Reviewers.docx

    pone.0348280.s006.docx (24.8KB, docx)

    Data Availability Statement

    All relevant data are within the paper and its Supporting Information files.


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