Diagnostics of animal rabies by enzyme immunoassay. Comparison of direct and indirect sandwich Elisa options

Sukharkov A.Yu., Nazarov N.A., Metlin A.E.

Summary

Diagnostics of rabies is important to provide timely post-exposure medical care for people and rabies control among animals. Currently, basic diagnostic methods of rabies are: reaction of immunofluorescence and biological test in mice, as well as virus isolation in cell culture. The main disadvantage of biological tests in mice and virus isolation in cell culture is duration of the study. The authors have developed a direct sandwich ELISA option for diagnostics of rabies. Developed solid-phase direct sandwich enzyme immunoassay option for the diagnostics of animal rabies, based on polyclonal antibodies to rabies virus ribonucleoprotein, and comparative trials of the direct method with the indirect sandwich ELISA variant, previously developed. Probation of both methods showed similar results in 100% of cases. Diagnostic sensitivity and specificity was respectively 97,14% and 100%, respectively. In case of equal diagnostic value of direct sandwich option has an advantage over indirect, as more and express less costly method.

Key words: rabies, rabies virus ribonucleoprotein, diagnostics of rabies, ELISA, brain, rabies peroxidate conjugate.

References

  1. Metlin A.E., Nazarov N.A., Rybakov S.S. Diagnostika beshenstva zhivotnykh s ispolzovaniem reaktsii immunofluorestsentsii: bibliografiya [Diagnostics of animal rabies using ELISA: bibliography]. – Veterinariya. – 2006. – pp. 20-23. – Print. (in Russ.).
  2. Nazarov N.A., Mihaylina N.M., Rybakov S.S. Razrabotka tverdofaznogo nepryamogo sendvich-varianta immunofermentnogo analiza dlya diagnostiki beshenstva zhivotnykh [Development of solid-phase indirect sandwich ELISA options for the diagnostics of rabies]. – Vladimir (2006): pp. 117-124. – Print. (in Russ.).
  3. Ponomaryov A.P., Nazarov N.A., Rybakov S.S. Sovershenstvovanie metoda ochistki i koncentrirovaniya virusa beshenstva [Improvement in the method of cleaning and concentration of rabies virus]. – Pokrov (2001): pp. 49-51. – Print. (in Russ.).
  4. Egorov A.M., Osipov A.P., Dzantiev B.B., Gavrilova E.M. Teoriya I praktika immunofermentnogo analiza [Theory and practice of ELISA]. – Moscow (1991). – 288 p. – Print. (in Russ.).
  5. Bourhy, H., Perrin P. Rapid rabies enzyme immunodiagnosis (RREID) for rabies antigen detection // Laboratory techniques in rabies. – 4th ed. – Geneva: World Health Organization, 1996. – P. 105-113.
  6. Meslin F.-X., Kaplan M.M. An overview of laboratory techniques in the diagnosis and prevention of rabies and in rabies research // Laboratory techniques in rabies. – 4th ed. – Geneva: World Health Organization, 1996. – P. 9-27.
  7. Nandy S. Recent advancement in the diagnosis of rabies //Asian livestock. – 1994. – Vol. 19, N°11. – P. 150-152.
  8. Perrin. P. Techniques for the preparation of rabies conjugates // Laboratory techniques in rabies. – 4th ed. – Geneva: World Health Organization, 1996. – P. 433-444.
  9. Protein measurement with Folin phenol reagent / O.H. Lowry, N.J. Rosebrough, A.L. Farr, R.J. Randall. // J. Biological Chemistry. – 1951. – Vol. 193. – P. 265-275.
  10. Singh H. β-Propiolactone-inactivated sheep brain vaccine // Laboratory techniques in rabies. – 4th ed. – Geneva: World Health Organization, 1996. – P. 234-242.
  11. Webster W.A., Casey G.A. Virus isolation in neuroblastoma cell culture // Laboratory techniques in rabies. – 4th ed. – Geneva: World Health Organization, 1996. – P. 96-103.

Author affiliation

Nazarov Nikolay A., Ph.D. in Biology, leading scientific researcher of the All-Russian Research Institute of Animal Health; e-mail: nazarov_na@arriah.ru.

Metlin Artem E., Ph.D. in Veterinary Medicine, laboratory manager of the All-Russian Research Institute of Animal Health (Federal Centre for Animal Health); e-mail: metlin@arriah.ru.

Responsible for correspondence with the editorial board: Sukharkov Andrey Yu., postgraduate student, leading biologist of the All-Russian Research Institute of Animal Health; mcrd. Yurevets, Vladimir, 600901; phone: (4922) 26-38-77, (4922) 26-06-14, (4922) 26-19-14; e-mail: andrey_33rus@mail.ru.

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