Table 4.

Modern (non-PCR/non-LAMP-based) methods used for malaria detection and evidence of use in developed countries.

Modern Methods	Specimen Used	Description	Invasive/Non-Invasive	Point of Care/Molecular/Other	Advantages	Disadvantages	Developed Countries	References
Malaria SD Bioline RDT kit	Urine, Saliva, Blood	Using immuno-chromatography to detect PfhRP2 and PLDH following manufacturer’s instructions	Non-invasive/Invasive	Point of care	Effective for non-invasive detection of malaria; low cost	Low sensitivity		[64]
Other (RDTs)	Blood	Immunochromatography/ according to manufacturer’s instructions	Invasive	Point of care	Suitable for point of care in hard-to-access areas; low cost	Low sensit-ivity for some kits; poor identification of non-falciparum infections for some brands	Indonesia Australia, USA	[14,15,17,18,65,66,67,68,69,70,71]
Computeri-zed/digital deep mach-ine learnin-g approach	Blood	Machine learning models are used to detect malaria parasites in blood smears. Some can be integrated into smartphone detection apps	Invasive	Other	Accurate/ reliable	For some, results are affected by quality of smears	USA, Taiwan, China, Turkey	[72,73,74,75,76,77,78,79,80,81]
Spectros-copy	Blood	Blood samples are analyzed using spectroscopy	Invasive	Other	Highly effective for identifying infected cell	Only qualitative results obtained	Thailand, China, Australia	[82,83,84]
Portable Optical Diagnostic System (PODS)	Blood	Works by differential optical spectroscopy. The change in optical power before and after a magnet is applied, is monitored in order to determine β-hematin concentration in whole blood	Invasive	Point of care	Portable; low cost; useful for low resource settings; high sensitivity	Not widely available	USA	[85]
Ultra bright SERS nanorattles	Blood	DNA detection method that uses sandwich hybridization of magnetic bead, target sequence, and ultrabright SERS nanorattle are employed	Invasive	Molecular/point of care	Sensitive; can be automated and added to portable devi-ces for POC diagnosis; can identify SNPs hence, discri-minate betw-een wild-type and mutant parasites	Not widely available	USA	[86]
Automated Microscopy/Digital Analysis	Blood	Comprises a fluorescent dye staining or Giemsa staining and an automated microscopy platform and digital analysis	Invasive	Other	Rapid diagn-osis and par-asite density monitoring. High sens- itivity, linear-ity, and precision	Not widely available	Korea, Finland, Sweden	[87,88,89]
Flow cytometry	Blood	Parasites are detected and quantified in blood by use of analyzers utilizing flow cytometry technology	Invasive	Molecular	Rapid and high sensiti-vity; useful for mass screening	May not be able to distinguish plasmodium species	Netherlands, France, USA, South Africa, Japan	[90,91,92,93,94]
Thin-Film Optical Filters	Blood	A thin film optical device is used based on optical reflectance spectrophotometry, for the parasite detection through haemozoin quantification	Invasive	Point of care	High sensitivity	High transmittance regions outside target wavelength	Portugal	[95]
Rotating cr- ystal magn-eto optical detection (RMOD) method	Blood	RMOD works by detection of the periodic modulation of light transmission. This is induced by hemozoin crystals which co-rotates with a rotating magnetic field	Invasive	Other	Higher sensitivity and accuracy than light microscopy	Sensitivity is poorer than PCR methods	Thailand, Hungary	[96,97,98]
Hemozin-Based Malaria diagnostic device (GazelleTM)	Blood	Using magneto-optical technology, the device detects hemozoin produced by Plasmodium	Invasive	Other	Sensitivities comparable to light micr-oscopy; faster than micros-copy; portab-le; can run on battery power	Unable to distinguish between species		[16]
Hemozoin-generated vapor nanobubbles	Blood vessel (transdermal)	Hemozoin generates a transient vapor nanobubble around hemozoin in response to a short and safe laser pulse. The acoustic signals of these nanobubbles that are malaria specific enable detection	Non-invasive	Point of care	Non-invasive; rapid	Not widely available	USA	[99]
Electroche-mical immunosensor	Blood	Egg yolk IgY antibodies against Plasmodium vivax lactate dehydrogenase antigen are immobilized on a gold electrode surface followed by differential pulse voltammetry and contact angle measurements are made.	Invasive	Point of care	High Sensitivity for malaria caused by P. vivax	Only malaria caused by P. vivax can be detected	Brazil	[100]
Simplified ELISA)/PfHRP 2 ELISA	Blood	Modified ElISA was performed on blood samples.	Invasive	Point of care	High sensitivity, portable and low cost	Not widely available	Spain UK Denmark	[101,102]
Multiple-xed ELISA based assay	Blood	Multiplexed ELISA-based (either planar-based array or magnetic bead-based platforms) technologies are used for malaria detection	Invasive	Molecular	Can detect malaria spe-cies mutants; have high throughput potential	Not widely available	USA	[103]
Dye-Cou-pledApt-amer-Capt-ured Enzy-me-Cataly-zed assay	Blood	Aptamer- and enzyme-based method is used to detect malaria infection in blood. Method can be used on instrument or instrument free platform	Invasive	Molecular/point of care	Low cost; useful for resource-limited and point-of-care settings.	Not widely available		[104]
Recombinase-Aided Amplificat-ion with Lateral Flow Dip-stick Assay (RAA-LFD)	Blood	A combination of recombinase-aided amplification lasting for 15 min at 37 degrees and lateral flow dipstick is used to detect plasmodium species in blood	Invasive	Molecular/point of care	Highly sensitive, specific, low cost, convenient for on-site screening and low resource settings.	Not widely available	China	[105]
Portable image-based Cytometer	Blood	P. falciparum-infected blood cells are identified and counted from Giemsa-stained smears using the image based portable cytometer.	Invasive	Other	Simple to operate; low cost	Not widely available	Singapore	[106]
Two-stage sample-to-answer sy-stem based on nucleic acid ampl-ification approach	Blood	It combines the dimethyl adipimidate (DMA)/thin film sample processing (DTS) technique and the Mach–Zehnder interferometer isothermal solid-phase DNA amplification (MZI-IDA) technique to detect infection in blood	Invasive	Molecular	High sensitivity, rapid	Not widely available	Singapore, Korea	[107]
Fluorescen-ce In Situ Hybridization (FISH) Assays	Blood	Detects and localizes specific malaria nucleic acid sequences by hybridizing with complementary sequences that are labeled with fluorescent probes	Invasive	Molecular	High sensitivity	Skilled expertise required.	USA	[108,109]
NMR-based hemozoin detection	Blood	Detection is based on the ability to recognize the paramagnetic susceptibility of malaria hemozoin crystals	Invasive	Molecular/point of care	High sensitivity and rapid	Not widely available	Australia, Singapore, USA	[110,111,112]
Multi-omics	Varies	Integrating data from different omic methods	Invasive/non-invasive	Other	Comprehen-sive underst-anding of the infection	Requires skilled experitise	Austria USA Columbia	[113,114,115,116]