Method for simultaneous detection of serological dengue fever markers

Abstract

The aim of the work was to develop and test a method for the simultaneous detection of dengue fever markers (virus-specific IgM, IgG antibodies and NS1 antigen) in blood samples.

Material and methods. Detection of specific markers of dengue fever in samples of commercial verification panels was performed by dot-immunoassay, immune chromatography commercial test and the developed method.

Results and discussion. A method of dot-immunoassay on flat protein array was developed. The method included two components to obtain the signal: Gold nanoparticle–protein conjugate and a two-component development system (silver nitrate solution and a mixture of metоl and citric acid) with enhancement of optical signal with an thiourea alkaline solution. This technique simultaneously detects the NS1 protein of dengue virus and specific IgM and IgG in blood samples. The analysis takes for 62 minutes at a temperature of 20–30 °C. An experimental kit was created based on the method.

Conclusion. The developed kit can be used for individual analysis and for mass screening of clinical samples. Evaluation of the method on a samples of verification panels showed that the results were consistent with commercial tests, χ² 0.705–0.047. The method can be used for analysis of clinical samples, both in the laboratory and out of it.

Keywords:dengue fever; serological markers, simultaneous detection, multiplex dot immunoassay, protein matrices, colloidal gold

Funding. The study was carried out as part of the fulfilment of the state assignment of the Federal Service for the Protection of Consumer Rights and Human Welfare.

Conflict of interest. The authors confirm that there is no conflict of financial/non-financial interests related to the writing of the article.

Contribution. Conceptualization, methodology, writing, project administration – Poltavchenko A.G.; methodology, investigation, validation – Ersh A.V.; investigation, formal analysis, editing, data curation – Filatov P.V.; resources, methodology, investigation – Bayandin R.B.; resources, methodology, investigation, editing – Ushkalenko N.D.

For citation: Poltavchenko A.G., Ersh A.V., Filatov P.V., Bayandin R.B., Ushkalenko N.D. Method development for simultaneous detection of serological dengue fever markers. Infektsionnye bolezni: novosti, mneniya, obuchenie [Infectious Diseases: News, Opinions, Training]. 2023; 12 (1): 75–83. DOI: https://doi.org/10.33029/2305-3496-2023-12-1-75-83 (in Russian)

References

1. Owino E. Aedes spp mosquitoes and emerging neglected diseases of Kenya. Int J Mosq Res. 2018; 5: 1–11.

2. Dengue and severe dengue. URL: https://www.who.int/news-room/fact-sheets/detail/dengue-and-severe-dengue (date of access December 26, 2022).

3. St John A.L., Gubler D.J. Dengue Viruses (Flaviviridae). In: D.H. Bamford, M. Zuckerman (eds). Encyclopedia of Virology. 4th ed. Oxford: Academic Press, 2021: 218–31. ISBN 978-0-12-814516-6.

4. World Health Assembly 58. Revision of the International Health Regulations. World Health Organization, 2005. URL: https://apps.who.int/gb/ebwha/pdf_files/WHA58/WHA58_3-en.pdf (date of access December 26, 2022).

5. Ternovoi V.A., et al. Identification of dengue infection markers in patients after visiting to dengue endemic countries. Molekulyarnaya genetika, mikrobiologiya i virusologiya [Molecular Genetics, Microbiology, Virology]. 2019; 37 (3): 140–8. DOI: https://doi.org/10.17116/molgen201937031140 (in Russian)

6. Nechaev V.V., et al. Clinical-epidemiological characteristics delivery cases of tropic dengue fever. Zhurnal infektologii [Journal of Infectology]. 2021; 13 (1): 78–85. DOI: https://doi.org/10.22625/2072-6732-2021-13-1-78-85 (in Russian)

7. On the state of sanitary and epidemiological well-being of the population in the Russian Federation in 2021: State report. Moscow: Federal’naya sluzhba po nadzoru v sfere zashchity prav potrebiteley i blagopoluchiya cheloveka, 2022: 340 p. URL: https://rospotrebnadzor.ru/documents/details.php?ELEMENT_ID=21796 (date of access December 26, 2022). (in Russian)

8. Ganushkina L.A., et al. Detection of Aedes aegypti, Aedes albopictus, and Aedes koreicus in the Area of Sochi, Russia. Vector Borne Zoonotic Dis. 2016; 16 (1): 58–60. DOI: https://doi.org/10.1089/vbz.2014.1761

9. Knipe D.M., Howley P. Fields Virology. 6th ed. Philadelphia: Lippincott Williams & Wilkins, 2013: 2664 p. ISBN 978-1-4698-3066-7.

10. Kuhn R.J., et al. Structure of dengue virus: implications for flavivirus organization, maturation, and fusion. Cell. 2002; 108 (5): 717–25. DOI: https://doi.org/10.1016/s0092-8674(02)00660-8

11. Darwish N., Alias Y., Khor S. An introduction to dengue-disease diagnostics. Trends Anal Chem. 2015; 67: 45–55. DOI: https://doi.org/10.1016/j.trac.2015.01.005

12. Hue K.D.T., et al. Validation of an internally controlled one-step real-time multiplex RT-PCR assay for the detection and quantitation of dengue virus RNA in plasma. J Virol Methods. 2011; 177 (2): 168–73. DOI: https://doi.org/10.1016/j.jviromet.2011.08.002

13. Tandel K., et al. Detection of dengue virus serotypes by single-tube multiplex RT-PCR and multiplex real-time PCR assay. Med J Armed Forces India. 2022; 78 (3): 333–8. DOI: https://doi.org/10.1016/j.mjafi.2021.09.001

14. Akinshina Y.A., et al. Comparison of the application of domestic «ELISA-IGM-DENGUE» kit, delivered in the D.I. Ivanovsky institute of virology (Moscow, Russian Federation) and «ANTI-DENGUE VIRUS ELISA IGM» KIT (EUROIMMUN, Germany) for the serodiagnosis of Dengue fever. Epidemiologiya i infektsionnye bolezni [Epidemiology and Infectious Diseases]. 2017; 22 (1): 4–8. DOI: https://doi.org/10.17816/EID40949 (in Russian)

15. Poltavchenko A.G., et al. The selection and optimization of the detection system for self-contained multiplexed dot-immunoassay. J Immunoassay Immunochem. 2016; 37 (5): 540–54. DOI: https://doi.org/10.1080/15321819.2016.1174134

16. Ushkalenko N., et al. Evaluation of Rapid dot-immunoassay for detection orthopoxviruses using laboratory-grown viruses and animal’s clinical specimens. Viruses. 2022; 14 (11): 2580. DOI: https://doi.org/10.3390/v14112580

17. Zhang H., et al. NS1-based tests with diagnostic utility for confirming dengue infection: a meta-analysis. Int J Infect Dis. 2014; 26: 57–66. DOI: https://doi.org/10.1016/j.ijid.2014.02.002

18. Weiskopf D., et al. Immunodominance changes as a function of the infecting dengue virus serotype and primary versus secondary infection. J Virol. 2014; 88 (19): 11 383–394. DOI: https://doi.org/10.1128/JVI.01108-14

19. Poltavchenko A.G., et al. The selection of system of detection for multiplex dot-immuneanalysis of antibodies. Klinicheskaya laboratornaya diagnostika [Clinical Laboratory Diagnostics]. 2016; 61 (4): 229–33. DOI: https://doi.org/10.18821/0869-2084-2016-61-4-229-233 (in Russian)

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CHIEF EDITOR
Aleksandr V. Gorelov
Academician of the Russian Academy of Sciences, MD, Head of Infection Diseases and Epidemiology Department of the Scientific and Educational Institute of Clinical Medicine named after N.A. Semashko ofRussian University of Medicine, Ministry of Health of the Russian Federation, Professor of the Department of Childhood Diseases, Clinical Institute of Children's Health named after N.F. Filatov, Sechenov First Moscow State Medical University, Ministry of Health of the Russian Federation, Deputy Director for Research, Central Research Institute of Epidemiology, Rospotrebnadzor (Moscow, Russian Federation)

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