A verified multiplexed immunoassay for detecting antibodies against infectious pathogens

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Baseline screening  of population groups at increased risk of developing infectious diseases (ID) includes a number of ELISA-based analyses, thereby accounting for high cost laborious and time-consuming examination. Earlier wedeveloped a cheap multiplex assay allowing to quickly perform a complex initial testing. Here, we would like to describe a novel kit designed to simultaneously detect antibodies against the six ID pathogens in blood products (plasma, serum etc.): human immunodeficiency virus, hepatitis B and C viruses, cytomegalovirus, Treponema pallidum and Toxoplasma gondii. The kit is based on multiplex dot-immunoassay over flat protein arrays (immune chips) utilizing colloidal gold conjugates and silver development. It allows carrying out complex analysis at room temperature within 70 minutes and does not require highly-qualified personnel. In this article, the results of laboratory tests of the experimental sample of a multiplex kit are compared with commercial ELISA kits from five Russian manufacturers using an array of 240 blood product samples including 200 clinical serum (plasma) samples from patients with ID and 40 plasma samples from donors. It was shown that the multiplex test analysis has a sensitivity and specificity of at least 95%, and optical signals of the dot-assay correlate well with the optical density values obtained using commercial ELISA kits and allow a semiquantitative evaluation of the content of specific antibodies in the sample. Furthermore, multiplex analysis is quicker and cheaper compared to ELISA and can be carried out in field. The kit for multiplex dot-immunoassay of antibodies can provide a complex approach to the diagnosis of ID, significantly simplify initial testing, make it faster, more efficient and affordable for patients.

About the authors

A. G. Poltavchenko

State Research Center of Virology and Biotechnology “Vector”

Email: poltav@vector.nsc.ru

Poltavchenko Alexander Georgievich - PhD, MD (Biology), Head of Immunochemistry Laboratory, State Research Center of Virology and Biotechnology “Vector”.

630559, Novosibirsk Region, Koltzovo.

Phone/fax: +7 (383) 336-74-09 (office); +7 (913) 735-91-74 (mobile).

SPIN: 9153-2935

Russian Federation

O. V. Nechitaylo

State Research Center of Virology and Biotechnology “Vector”

Email: nechitaylo_ov@vector.nsc.ru

Nechitaylo Oleg Vyacheslavovich - Trainee Researcher, Immunochemistry Laboratory, State Research Center of Virology and Biotechnology “Vector”.

630559, Novosibirsk Region, Koltzovo.

Russian Federation

P. V. Filatov

State Research Center of Virology and Biotechnology “Vector”

Email: medic@sibmail.ru
ORCID iD: 0000-0001-7763-3808

Filatov Pavel Vladimirovich - PhD (Biology), Researcher, Immunochemistry Laboratory, State Research Center of Virology and Biotechnology “Vector”.

630559, Novosibirsk Region, Koltzovo.

SPIN: 2055-2826

Russian Federation

A. V. Ersh

State Research Center of Virology and Biotechnology “Vector”

Author for correspondence.
Email: ersh_av@vector.nsc.ru

Ersh Anna Vasilievna - PhD (Biology), Researcher, Immunochemistry Laboratory, State Research Center of Virology and Biotechnology “Vector”.

630559, Novosibirsk Region, Koltzovo.

Russian Federation


  1. Ляужева Ф.М., Тленкопачев Р.С., Жибурт Е.Б. Выбраковка доноров по маркерам инфекций в Кабардино-Балкарской Республике // Трансфузиология. 2014. № 4. С. 32–36.
  2. Полтавченко А.Г., Ерш А.В., Пьянков С.А., Никонов А.М., Волков Г.Н, Кривенчук Н.А. Многопрофильная серодиагностика инфекционных заболеваний. Инструментальный учет результатов анализа // Биотехнология. 2013. № 4. С. 74–82.
  3. Burgess S.T.G., Kenyon F., O’Looney N., Ross A.J., Kwan M.C., Beattie J.S., Petrik J., Ghazal P., Campbell J.C. A multiplexed protein microarray for the simultaneous serodiagnosis of human immunodeficiency virus/hepatitis C virus infection and typing of whole blood. Anal. Biochem., 2008, vol. 382, pp. 9–15. doi: 10.1016/j.ab.2008.07.017
  4. Corstjens P.L.A.M., Chen Z., Zuiderwijk M., Bau H.H., Abrams W.R., Malamud D., Niedbala R.S., Tanke H.J. Rapid assay format for multiplex detection of humoral immune responses to infectious disease pathogens (HIV, HCV, and TB). Ann. N.Y. Acad. Sci., 2007, vol. 1098, pp. 437–445. doi: 0.1196/annals.1384.016
  5. Ellington A.A., Kullo I.J., Bailey K.R., Klee G.G. Antibody-based protein multiplex platforms: technical and operational challenges. Clin. Chem., 2010, vol. 56, pp. 186–194. doi: 10.1373/clinchem.2009.127514
  6. Hartmann M., Roeraade J., Stoll D., Templin M.F., Joos T.O. Protein microarrays for diagnostic assays. Anal. Bioanal. Chem., 2009, vol. 393, pp. 1407–1416. doi: 10.1007/s00216-008-2379-z
  7. Len O., Garzoni C., Lumbreras C., Molina I., Meije Y., Pahissa A., Grossi P. Recommendations for screening of donor and reci- pient prior to solid organ transplantation and to minimize transmission of donor-derived infections. Clin. Microbiol. Infect., 2014, vol. 20, iss. 7, pp. 10 –18. doi: 10.1111/1469-0691.12557
  8. Lochhead M.J., Todorof K., Delaney M., Ives J.T., Greef C., Moll K., Rowley K., Vogel K., Myatt C., Zhang X.Q., Logan C., Benson C., Reed S., Schooley R.T. Rapid multiplexed immunoassay for simultaneous serodiagnosis of HIV-1 and coinfections. J. Clin. Microbiol., 2011, vol. 49, pp. 3584–3590. doi: 10.1128/JCM.00970-11
  9. Myyrylainen T., Talha S.M., Swaminathan S., Vainionpaa R., Soukka T., Khanna N., Pettersson K. Simultaneous detection of human immunodeficiency virus 1 and hepatitis B virus infections using a dual-label time-resolved f luorometric assay. J. Nanobiotechnol., 2010, vol. 8, pp. 27–32. doi: 10.1186/1477-3155-8-27
  10. Petrik J. Diagnostic applications of microarrays. Transfus. Med., 2006, vol. 16, pp. 233–247. doi: 10.1111/j.1365-148.2006.00673.x
  11. Poltavchenko A.G., Nechitaylo O.V., Filatov P.V., Ersh A.V., Gureyev V.N. Multiplex method for initial complex testing of antibodies to blood transmitted diseases agents. J. Virol. Meth., 2016, vol. 236, pp. 231–236. doi: 10.1016/j.jviromet.2016.08.003
  12. Poltavchenko A.G., Zaitsev B.N., Ersh A.V., Korneev D.V., Taranov O.S., Filatov P.V., Nechitaylo O.V. The selection and optimization of the detection system for self-contained multiplexed dot-immunoassay. J. Immunoassay Immunochem., 2016, vol. 37, pp. 540 –554. doi: 10.1080/15321819.2016.1174134
  13. Talha S.M., Saviranta P., Hattara L., Vuorinen T., Hytönen J., Khanna N., Pettersson K. Array-in-well platform-based multiplex assay for the simultaneous detection of anti-HIV- and treponemal-antibodies, and hepatitis B surface antigen. J. Immunol. Meth., 2016, vol. 429, pp. 21–27. doi: 10.1016/j.jim.2015.12.007
  14. Tighe P.J., Ryder R.R., Todd I., Fairclough L.C. ELISA in the multiplex era: potentials and pitfalls. Proteomics Clin. Appl., 2015, vol. 9, pp. 406–422. doi: 10.1016/j.jim.2015.12.007
  15. Wong J., Sibani S., Lokko N.N., LaBaer J., Anderson K.S. Rapid detection of antibodies in sera using multiplexed self-assembling bead arrays. J. Immunol. Meth., 2009, vol. 350, pp. 171–182. doi: 10.1016/j.jim.2009.08.013

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Copyright (c) 2019 Poltavchenko A.G., Nechitaylo O.V., Filatov P.V., Ersh A.V.

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