«IZVESTIYA IRKUTSKOGO GOSUDARSTVENNOGO UNIVERSITETA». SERIYA «BIOLOGIYA. ECOLOGIYA»
«THE BULLETIN OF IRKUTSK STATE UNIVERSITY». SERIES «BIOLOGY. ECOLOGY»
ISSN 2073-3372 (Print)

List of issues > Series «Biology. Ecology». 2021. Vol 37

Obtaining and Efficiency Assessment of Diagnostic Agglutinating Serum for Identifying the Causative Agent of Listeriosis

Author(s)
N. M. Khaptanova, N. M. Andreevskaya, Zh. A. Konovalova, N. G. Gefan, S. V. Luk'yanova, A.S. Ostyak, N. N. Kartsev, V. N. Borzenkov, S. V. Balakhonov
Abstract
Listeriosis is a saprozoonotic natural anthropurgic infectious disease caused by Listeria monocytogenes, with multiple pathways and factors of its transmission, characterized by pronounced clinical polymorphism from carriage and subclinical to severe generalized forms, meningitis and meningoencephalitis, and high mortality among newborn children and individuals with immunodeficiency. Express diagnostics of listeriosis is based on the use of immuno-chemical (immunodiffusion reaction, enzyme immunoassay) and molecular genetic (polymerase chain reaction) methods. In the practice of laboratory diagnostics, serological methods for the study of listeriosis remain in high demand for verifying the clinical diagnosis. One of the specific, reliable and available serological methods for laboratory diagnostics of causative agents of infectious diseases remains the agglutination test (AT), to conduct which, listeria agglutinating serum is required. Obtaining diagnostic agglutinating listeria serum with a high level of diagnostic sensitivity and specificity will allow timely identification of the pathogen in the test tube AT and on glass, based on the specific interaction of the antigen with the anti-body, with the formation of agglutinate visible to the naked eye. In this study a scheme has been developed for immunization of animal producers to obtain diagnostic listeria agglutinat-ing serum. Evaluation of the clinical efficiency of two series of the serum was carried out based on the indices of diagnostic sensitivity with different serovariants of L. monocytogenes and diagnostic specificity with closely related and heterologous strains in the test tube AT and on glass. During the study of 25 strains of L. monocytogenes serogroups I and II (serotypes 1/2a, 1/2b, 1/2c, 3a, 3b, 3c, 4a, 4b, 4c, 7) isolated on the territory of the Russian Federation from various sources (clinical material, food products, wastewater), the serum titer did not differ and amounted to 1:800 in the course of in vitro agglutination reaction; a positive result was obtained on glass in 100% of cases. The assessment of the statistical reliability of the ob-tained test results was at least 89 % with a statistical significance of 95 %, which indicates a high diagnostic efficiency of the serum. In order to establish intra-stage convergence and inter-series reproducibility of studies, two probabilities of binomial distributions were compared. The convergence of staged studies for all positive samples has been proven. Based on the re-sults obtained, listeriosis serum can be used as a medical preparation for in vitro diagnostics of the causative agent of listeriosis.
About the Authors

Khaptanova Natalya Markelovna, Junior Research Scientist, Irkutsk Antiplague Research Institute of Siberia and Far East, 78, Trilisser St., Irkutsk, 664047, Russian Federation, e-mail: khaptanchik@mail.ru

Andreevskaya Nina Mikhailovna, Candidate of Sciences (Biology), Senior Research Scientist, Irkutsk Antiplague Research Institute of Siberia and Far East,78, Trilisser St., Irkutsk, 664047, Russian Federation, e-mail: andreevskayа45@mail.ru

Konovalova Zhanna Anatol'evna, Candidate of Sciences (Biology), Senior Research Scientist, Irkutsk Antiplague Research Institute of Siberia and Far East. 78, Trilisser St., Irkutsk, 664047, Russian Federation, e-mail: adm@chumin.irkutsk.ru

Gefan Natalya Gennadyevna, Candidate of Sciences (Medicine), Head of Department, Irkutsk Antiplague Research Institute of Siberia and Far East, 78, Trilisser St., Irkutsk, 664047, Russian Federation, e-mail: adm@chumin.irkutsk.ru

Luk'yanova Svetlana Vladimirovna, Candidate of Sciences (Biology), Research Scientist, Irkutsk Antiplague Research Institute of Siberia and Far East, 78, Trilisser St., Irkutsk, 664047, Russian Federation, e-mail: svetalukyan@mail.ru

Ostyak Aleksandr Sergeevich, Research Scientist, Irkutsk Antiplague Research Institute of Siberia and Far East, 78, Trilisser St., Irkutsk, 664047, Russian Federation, e-mail: ostyakalex@mail.ru

Kartsev Nikolay Nikolaevich, Candidate of Sciences (Medicine), Senior Research Scientist State Research Center for Applied Microbiology and Biotechnology, 24, “Kvartal A” Territory, Obolensk setl., Moscow region, 142279, Russian Federation, e-mail: kartsev@obolensk.org

Borzenkov Valerii Nikolaevich, Candidate of Sciences (Biology), Senior Research Scientist, State Research Center for Applied Microbiology and Biotechnology 24, “Kvartal A” Territory, Obolensk setl., Moscow region, 142279, Russian Federation, e-mail: vbn5314@mail.ru

Balakhonov Sergei Vladimirovich, Doctor of Sciences (Medicine),Professor, Director, Irkutsk Antiplague Research Institute of Siberia and Far East, 78, Trilisser St., Irkutsk, 664047, Russian Federation, e-mail: balakhonov.irk@mail.ru

For citation
Khaptanova N.M., Andreevskaya N.M., Konovalova Zh.A., Gefan N.G., Luk'yanova S.V., Ostyak A.S., Kartsev N.N., Borzenkov V.N., Balakhonov S.V. Obtaining and Efficiency Evaluation of Diagnostic Agglutinating Serum for Identifying the Causative Agent of Listeriosis. The Bulletin of Irkutsk State University. Series Biology. Ecology, 2021, vol. 37, pp. 43-53. https://doi.org/10.26516/2073-3372.2021.37.43 (in Russian)
Keywords
Listeria monocytogenes, listeria agglutinating serum, diagnostic sensitivity, diag-nostic specificity, diagnosis.
UDC
579.869.1+616-078
DOI
https://doi.org/10.26516/2073-3372.2021.37.43
References

Efimochkina N.R. Mikrobiologiya pishchevykh produktov i sovremennye metody detektsii patogenov [Microbiology of food products and modem methods of detection of pathogens]. Moscow, RAMS Publ., 2013, 518 p. (in Russian)

Metodicheskie rekomendatsii po poryadku provedeniya ekspertizy kachestva, effektivnosti i bezopasnosti meditsinskikh izdelii dlya gosudarstvennoi registratsii[Methodological recommendations on the procedure for conducting an examination ofthe quality, effectiveness and safety of medical devices for state registration]. Moscow, VNIIIMT Publ., 2018. 158 p. (in Russian)

Peretrukhina A.T., Blinova E.I. Bakteriinye i virusnye preparaty [Bacterial and viral drugs]. Moscow, Akademiya Estestvoznaniya Publ., 2011, 311 p.

Tartakovskij I.S. Listerii: rol v infektsionnoi patologii cheloveka i laboratornaya diag-nostika [Listeria: role in human infectious pathology and laboratory diagnostics].Klinicheskaya mikrobiologiya i antimikrobnaya khimioterapiya [Clinical Microbiology and Antimicrobial Chemotherapy], 2000, vol. 2, no. 2, pp. 20-30. (in Russian)

Khaptanova N.M., Lukyanova S.V., Kuznetsov V.I., Gefan N.G., Аndreevskaya N.M., Konovalova Zh.A., Ostyak А.S., Kosilko V.S. Designing a nutrient medium for the accumulation of microbial mass of Listeria. Acta Biomedica Scientifica, 2020, vol. 5, no. 4, pp. 60-66. (in Russian) https://doi.org/10.29413/ABS.2020-5.4.8

Chastnaya meditsinskaya mikrobiologiya s tekhnikoi mikrobiologicheskikh issledovanii [Partial medical microbiology with the technique of microbiological research]. Labinskaya A.S., Blinkova L.P., Eshchina A.S., Ankirskaya A.S. (Eds.). St.-Petersb., Lan’ Publ., 2020, 608 p. (in Russian)

Ricci A., Allende A., Bolton D., Chemaly M., Davies R., Fernández Escámez P. S., Gi-rones R., Herman L., Koutsoumanis K., Nørrung B., Robertson L., Ru G., Sanaa M., Simmons M., Skandamis P., Snary E., Speybroeck N., Ter Kuile B., Threlfall J., Wahlström H., Tak-kinen J., Wagner M., Arcella D., Da Silva Felicio M. T., Georgiadis M., Messens W., Lind-qvist R. Listeria monocytogenes contamination of ready-to-eat foods and the risk for human health in the EU. EFSA journal, 2018, vol. 16, no. 1, 170 p. https://doi.org/10.2903/j.efsa.2018.5134

Matle I., Mbatha K. R., Madoroba E. A review of Listeria monocytogenes from meat and meat products: Epidemiology, virulence factors, antimicrobial resistance and diagnosis. Onderstepoort J. Vet Res., 2020, vol. 87, no. 1, pp. 1-20. https://doi.org/10.4102/ojvr.v87il.1869

Lowry O. H., Rosebrough N. J., Farr A. L., Randall R. J. Protein measurement with the Folin phenol reagent. J. Biol. Chem., 1951, vol. 193, no. 1, pp. 265-275. https://doi.org/10.1016/S0021-9258(19)52451-6

Koutsoumanis K., Alvarez-Ordonez A., Bolton D., Bover-Cid S., Chemaly M., Davies R., De Cesare A., Herman L., Hilbert F., Lindqvist R., Nauta M., Peixe L., Ru G., Simmons M., Skandamis P., Suffredini E., Jordan K., Sampers I., Wagner M., Da Silva Felicio M. T., Georgiadis M., Messens W., Mosbach-Schulz O., Allende A. The public health risk posed by Listeria monocytogenes in frozen fruit and vegetables including herbs, blanched during processing. EFSA Journal, 2020, vol. 18, no. 4, 101 p. https://doi.org/10.2903/j.efsa.2020.6092


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