Objective of research: to estimate the efficacy of fractionated antigen of mature cattle parasites Setaria labiato-papillosa in diagnosisof dirofilariasis (Dirofilaria immitis). Materials and methods: Antigens of mature Setaria labiato-papillosa, reference sera frominfected and not infected dogs. Reaction of immunodiffusion (RID) in agar gel using reference sera from dogs infected with D. immitis, and extracts from S. labiato-papillosa and D. immitis. Gel-chromatographic method on Pharmacia columns of Superose 12 Prep Grade and immunoenzymatic reaction (IER). Results and discussion: The reaction of immunodiffusion (RID) was used to detectcrossreacting antigens of D. immitis and S. labiato-papillosa. The fractionation of antigen-extract from S. labiato-papillosa allows to get 40 protein fractions, 7 (17,5%) of which were of interest for diagnosis of dirofilariasis. The evaluation of these fractions with respect to diagnosis of dirofilariasis was performed by immunoenzymatic reaction with 47 serum samples from dogs including 11 serum samples from D. immitis infected dogs; 6 — D. repens; 3 — Babesia canis; 3 — Toxocara canis; 9 — samples from dogs with non-parasitic pathology and 15 — from clinically healthy dogs.These results revealed that in one of 11 serum samples infected by D. immitis, and in two of 6 serum samples infected by D. repens — a false-negative response has been registered. The test sensitivity was 82,4%, the specificity — 83,3%. The sensitivity of the similar test with these sera using the purified antigen extract from mature D. immitis, was also 82,4%, and the specificity — 86,7%. The comparability of diagnostic test results for dirofilariasis by IER using antigenic preparations from Setaria and Dirofilaria allows to come to a conclusion that it is possible to use S. labiatopapillosa as a source of antigen for diagnosis of dirofilariasis (D. immitis).
Setaria labiato-papillosa, Dirofilaria immitis, cattle, dog sera, fractionated antigen, immunoenzymatic reaction.
Введение
За последние годы произошло значительное расширение ареала распространения дирофилярий, что привело к росту заболеваемости человека и животных дирофиляриозом и явилось следствием неослабевающего научного интереса к изучению этой медико- ветеринарной проблемы [6-10, 12, 19, 20].
Прижизненная диагностика дирофиляриоза, возбудителями которой являются D. immitis и D. repens, как правило, базируется на обнаружении микрофилярий в крови и их идентификации. Но поскольку дирофиляриоз продолжительное время может протекать бессимптомно, особенно при низкой интенсивности инвазии и соответственно низкой микрофиляриемии, то эти методы фактически не дают должного эффекта.
В таких случаях особый смысл приобретают исследования по разработке и использованию иммунодиагностических тестов, основу которых составляет выявление циркулирующих антигенов или антител, синтезируемых в организме инвазированного животного или человека.
1. Astaf’ev B.A., Petrov O.E. Evolutionary and genetic theory of parasitism. Uspehi sovremennoj biologii [Advances in modern biology], 1992, vol.112, no. 2, pp. 163-175. (In Russian)
2. Berezhko V.K., Khaydarov V.K., Dahno N.S., Shkurka E.P. Immunochemical analysis of somatic extract from mature Dirofilariaimmitis. Mater. dokl. nauch. konf. Vseros. o-va gel’mintol. RAN «Teoriya i praktika bor’by s parazitarnymi boleznyami». [Proc. of sci. conf. of All-Russian Society of Helminthol. RAS «Theory and practice of the struggle against parasitic diseases]. M., 2008, i. 9, pp. 69-73. (In Russian)
3. Berezhko V.K., Napisanova L.A., Medvedev A.Yu.,Khaydarov K.A., Shinkarenko A.N., Kolesnikov P.V. Physicochemical and immunodiagnostic characteristics of whole and fractionated somatic extract of Dirofilariaimmitis. Doklady Rossel’hozakademii. [Proceedings of the Russian Academy of Agricultural Sciences], 2014, no. 1, pp. 71-74. (In Russian)
4. Beskrovnaya Yu.G., Nagornyi S.A. Identification of microfilaria Dirofilaria spp. by polymerase chain reaction (PCR). Mater. dokl. nauch. konf. Vseros. o-va gel’mintol. RAN «Teoriya i praktika bor’by s parazitarnymi boleznyami». [Proc. of sci. conf. of All-Russian Society of Helminthol., RAS «Theory and practice of the struggle against parasitic diseases]. M., 2008, pp. 73-75. (In Russian)
5. Beskrovnaya Ju.G., Nagornyi S.A., Vaserin Yu.I. Efficacy evaluation of diagnosis of dirofilarias is indogsusing the Dirofilariarepens somatic antigen. Mater. dokl. nauch. konf. Vseros. o-vagel’mintol. RAN «Teoriya i praktika bor’by s parazitarnymi boleznyami». [Proc. of sci. conf. of All-Russian Society of Helminthol., RAS «Theory and practice of the struggle against parasitic diseases]. M., 2009, i. 10, pp. 61-63. (In Russian)
6. Krivorotova E.Yu. Biologicheskie aspekty dirofilyarioza v ryade sub’ektov Rossiyskoy Federatsii. Dis. … kand. biol. nauk [Biological aspects of dirofilariasis in a number of constituent territories of the Russian Federation. PhD diss... biol. sci.]. M., 2015. 141 p. (In Russian)
7. Medvedev A.Yu. Rasprostranenie dirofilyarioza sobak v Krasnodarskom krae i razrabotka ego diagnostiki immunofermentnoy reakciey. Diss. … kand. Biol. Nauk [Prevalence of dirofilariasis in dogs in Krasnodar Krai and development of diagnostic techniques using an immunoenzymatic reaction. PhD diss... biol. sci.]. M., 2007.137 p. (In Russian)
8. Sergiev V.P., Supryaga V.G., Darchenkova N.N., Zhukova L.A., Ivanova T.N. Human dirofilariasis in Russia. Rossiyskiy parazitologicheskiy zhurnal [Russian Journal of Parasitology], 2012, no. 4, pp. 60-64. (In Russian)
9. Sergiev V.P., Supryaga V.G., Bronshtein A.M., Ganushkina L.A.,Rakova V.M., Morozov E.N., Fedyanina L.V., Frolova A.A., Morozova L.F., Ivanova I.B., Darchenkova N.N., Zhukova L.A. Results of the study on dirofilariasis in Russia. Meditsinskaya parazitologiya i parazitarnye bolezni [Medical Parasitology and Parasitic Diseases], 2014, no. 3, pp. 3-9. (In Russian)
10. Fis’ko M.A., Firsov N.F. Dirofilyarioz [Dirofilariasis], Rostov-on- Don, 2006. 110 p. (In Russian)
11. Khaydarov K.A., Berezhko V.K. Antigenic spectrum of somatic extracts from mature Setaria and Dirofilaria. Rossiyskiy parazitologicheskiy zhurnal [Russian Journal of Parasitology], 2009, no 1, pp. 68–74. (In Russian)
12. Akao N. Human dirofilariasis in Japan. Trop Med Health, 2011, vol. 39, no. 1. Suppl. 2, pp. 65-71.
13. Atwell R.B., Sheridan A.B., Baldock F.C. An evaluation of the Dirochek test for detection of Dirofilariaimmitis antigen in dogs. Aust. Vet. J., 1988, vol. 65, no. 5, pp. 161-162.
14. Capron A., Brygoo E.R., Afchain D. Apport de letude de la structure antigenique a la phylogenie des Helminthes. Bull. Mus. Nat. hist. natur. Zoo, 1972, vol. 55, pp. 877-885.
15. Collins G.H., Pope S.E. An evaluation of an ELISA test for the detection of antigens of Dirofilariaimmitis. Aust. Vet. J., 1987, vol. 64, no. 10, pp. 318-319.
16. Gioia G., Lecová L., Genchi M. et al. Highly sensitive multiplex PCR for simultaneous detection and discrimination of Dirofilariaimmitis and Dirofilariarepens in canine peripheral blood. Vet. Parasitol. , 2010, vol. 172, no. 1-2, pp. 160-163. 17. Latrofa M.S., Dantas-Torres F., Annoscia G. et al. A duplex real-time polymerase chain reaction assay for the detection of and differentiation between Dirofilariaimmitis and Dirofilariarepens in dogs and mosquitoes. Vet. Parasitol., 2012, vol. 185, no.2-4, pp. 181-185.
17. Latrofa M.S., Dantas-Torres F., Annoscia G. et al. A duplex real-time polymerase chain reaction assay for the detection of and differentiation between Dirofilaria immitis and Dirofilaria repens in dogs and mosquitoes // Vet. Parasitol. — 2012. — Vol. 185, N 2-4. — P. 181-185.
18. Lee A.C., Bowman D.D., Lucio-Forster A. et al. Evaluation of a new in-clinic method for the detection of canine heartworm antigen. Vet. Parasitol., 2011, vol. 177, no. 3-4, pp. 387-391.
19. Mc Call J.W., Genchi C., Kramer L.H. et al. Heartworm disease in animals and humans. Adv. Parasitol., 2008, vol. 66, pp. 193-285.
20. Morchón R., Carretón E., González-Miguel J., Mellado-Hernández I. Heartworm Disease (Dirofilariaimmitis) and Their Vectors in Europe — New Distribution Trends. Front. Physiol., 2012, vol. 3, Art. 196.
21. Renaud F. Biodiversite, genetiqueet evolution dans les systemeshotes-parasites. Bull. Soc. Zool. Ff., 1992, vol. 117, no.1, pp. 109-115.
22. Rivasi F., Boldorini R., Criante P. et al. Detection of Dirofilaria (Nochtiella) repens DNA by polymerase chain reaction in embedded paraffin tissues from two human pulmonary locations. APMIS, 2006, vol. 114, no. 7-8, pp. 567-574.
23. Sun M., Zhuo W., Guo S. et al. Serological survey of canine dirofilariosis in Chongqing, Kunming, Nanchang, Fuzhou, Guangzhou, Shenzhen, and Nanning in Southern China. Vet. Parasitol., 2012, vol. 185, no. 2-4, pp. 225-228.
24. Weil G.J., Malane M.S, Powers K.G. Detection of circulating parasite antigens in canine dirofilariasis by counterimmunoelectrophoresis. Am. J. Trop. Med. Hyg., 1984., vol. 33, no. 3, pp. 425-430.
25. — vyp. 10. — S. 61-63.
