Skip to main content
NCBI home page
Springer Nature - PMC COVID-19 Collection logoLink to Springer Nature - PMC COVID-19 Collection
. 2017 Dec 12;11(4):255–261. doi: 10.1007/s13206-017-1401-y

Direct buffer composition of blood pre-process for nucleic acid based diagnostics

YeJi Kim 1, Won-Nyoung Lee 1, Hyun Jin Yoo 1, Changyoon Baek 1, Junhong Min 1,✉
PMCID: PMC7097592  PMID: 32226588

Abstract

Recently, a variety of methods, so called “direct buffer”, have been developed to utilize nucleic acid in the blood for the measurement of infectious bacteria and virus without any equipment in the field. In here, we first investigated the individual and combinatory effects of candidate chemicals which might be composed of the direct buffer on the PCR inhibition reduction of main compositions in whole blood. The long and short PEGs, Na2SO4 and GuSCN were selected as representative kosmotropic and chaotropic salts, respectively. MgCl2 were chosen as divalent cation source and NaOH was used to control blood pH. The effect of common non-ionic biological detergent was tested with Triton X-100 and SDS (Sodium Dodecyl sulfate) was chosen as anionic detergent. These results could provide a foundation for the development of sample preparation solution in nucleic acid based diagnostic field. As a result, the direct buffer developed in this study was able to detect viruses with a concentration of 102 pfu/100 μL of whole blood by a very simple method.

Keywords: Whole blood, Adenovirus, Direct buffer, Nucleic acid, Sample preparation

References

  • 1.Jeong J.H., Kim T.K., Oh S.W., Choi E.Y. Fluorescence immunochip assay for thyroid stimulating hormone in whole blood. BioChip J. 2013;7:408–414. doi: 10.1007/s13206-013-7413-3. [DOI] [Google Scholar]
  • 2.Chen G.-H., et al. Isolating and Concentrating Rare Cancerous Cells in Large Sample Volumes of Blood by Using Dielectrophoresis and Stepping Electric Fields. BioChip J. 2014;8:67–74. doi: 10.1007/s13206-014-8201-4. [DOI] [Google Scholar]
  • 3.Solier C., Langen H. Antibody-based proteomics and biomarker research–Current status and limitations. J. Proteomics. 2014;14:774–783. doi: 10.1002/pmic.201300334. [DOI] [PubMed] [Google Scholar]
  • 4.Wolcott M.J. Advances in Nucleic Acid-Based Detection Method. Clin. Microbiol. Rev. 1992;5:370–386. doi: 10.1128/CMR.5.4.370. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 5.Stumpf F., et al. LabDisk with complete reagent prestorage for sample-to-answer nucleic acid based detection of respiratory pathogens verified with influenza A H3N2 virus. Lab Chip. 2016;16:199–207. doi: 10.1039/C5LC00871A. [DOI] [PubMed] [Google Scholar]
  • 6.Wang B., et al. Rapid and Sensitive Detection of Severe Acute Respiratory Syndrome Coronavirus by Rolling Circle Amplification. J. Clin. Microbiol. 2005;43:2339–2344. doi: 10.1128/JCM.43.5.2339-2344.2005. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 7.Al-Soud W.A., Jonsson L.J., Radstrom P. Identification and Characterization of Immunoglobulin G in Blood as a Major Inhibitor of Diagnostic PCR. J. Clin. Microbiol. 2000;38:345–350. doi: 10.1128/jcm.38.1.345-350.2000. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 8.Schrader C., Schielke A., Ellerbroek L., Johne R. PCR inhibitors–Occurrence, properties and removal. J. Appl. Mirobiol. 2012;113:1014–1026. doi: 10.1111/j.1365-2672.2012.05384.x. [DOI] [PubMed] [Google Scholar]
  • 9.Zhang Z., Kermekchiev M.B., Barnes W.B. Direct DNA Amplification from Crude Clinical Samples Using a PCR Enhancer Cocktail and Novel Mutants of Taq. J. Mol. Diagn. 2010;12:152–161. doi: 10.2353/jmoldx.2010.090070. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 10.AL-Soud W.D., Radstrom P. Purification and Characterization of PCR-Inhibitory Components in Blood Cells. J. Clin. Microbiol. 2001;39:485–493. doi: 10.1128/JCM.39.2.485-493.2001. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 11.Chung S.H., Baek C., Cong V.T., Min J. The microfluidic chip module for the detection of murine norovirus in oysters using charge switchable micro-bead beating. Biosens. Bioelectron. 2015;67:625–633. doi: 10.1016/j.bios.2014.09.083. [DOI] [PubMed] [Google Scholar]
  • 12.Hall A.T., Zovanyi A.M., Christensen D.R., Koehler J.W., Minogue T.D. Evaluation of Inhibitor-Resistant Real-Time PCR Methods for Diagnostics in Clinical and Environmental Samples. e73845: PLoS One 8; 2013. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 13.Tan S.C., Yiap B.C. DNA, RNA and Protein Extraction: The Past and The Present. J. Biomed. Biotechnol. 2009;2009:574398. doi: 10.1155/2009/574398. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 14.DeAngelis M.M., Wang D.G., Hawkins T.L. Solidphase reversible immobilization for the isolation of PCR products. Nucleic Acids Res. 1995;23:4742–4743. doi: 10.1093/nar/23.22.4742. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 15.Boom R., et al. Rapid and Simple Method for Purification of Nucleic Acids. J. Clin. Microbiol. 1990;28:495–503. doi: 10.1128/jcm.28.3.495-503.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 16.Bashford G., et al. Automated bead-trapping apparatus and control system for single-molecule DNA sequencing. Opt. Express. 2008;16:3445–3455. doi: 10.1364/OE.16.003445. [DOI] [PubMed] [Google Scholar]
  • 17.Cho Y.K., et al. One-step pathogen specific DNA extraction from whole blood on a centrifugal microfluidic device. Lab Chip. 2007;7:565–573. doi: 10.1039/b616115d. [DOI] [PubMed] [Google Scholar]
  • 18.Baek C., Kim H.Y., Na D., Min J. A microfluidic system for the separation and detection of E.coli O157: H7 in soil sample using ternary interactions between humic acid, bacteria, and a hydrophilic surface. Sens. Actuators B. Chem. 2015;208:238–244. doi: 10.1016/j.snb.2014.11.028. [DOI] [Google Scholar]
  • 19.Burckhardt J. Amplification of DNA from Whole Blood. PCR Methods Appl. 1994;3:239–243. doi: 10.1101/gr.3.4.239. [DOI] [PubMed] [Google Scholar]
  • 20.Al-Soud W.A., Radstrom P. Effect of Amplification Facilitators on diagnostic PCR in the Presence of Blood, Feces, and Meat. J. Clin. Microbiol. 2000;38:4463–4470. doi: 10.1128/jcm.38.12.4463-4470.2000. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 21.Radstrom P., Knutsson R., Wolffs P., Lovenklev M., Lofstrom C. Pre-PCR Processing. Mol. Biotechnol. 2004;26:133–146. doi: 10.1385/MB:26:2:133. [DOI] [PubMed] [Google Scholar]
  • 22.Zuo D., Xu Y., Xu W., Zou H. The influence of PEG molecular weight on morphologies and properties of PVDF asymmetric membranes. Chinese J. Polym. Sci. 2008;26:405–414. doi: 10.1142/S0256767908003072. [DOI] [Google Scholar]

Articles from Biochip Journal are provided here courtesy of Nature Publishing Group

RESOURCES