IMMUNE-ENZYME TEST SYSTEM FOR DETERMINING SPECIFIC ANTIBODIES TO MORAXELLA BOVOCULI BACTERIA IN BOVINE BLOOD SERUM

FIELD: veterinary medicine.SUBSTANCE: invention refers to veterinary microbiology and concerns an enzyme immunoassay test system for serological diagnosis of infectious keratoconjunctivitis in cattle and control of postvaccinal immunity. Disclosed is an immunoenzymometric test system for determining...

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Hauptverfasser: Spiridonov Anton Gennadevich, Makaev Kharis Nurtdinovich, Spiridonov Gennadij Nikolaevich, Kozulina Oksana Vladimirovna, Nikitin Andrej Ivanovich, Valebnaya Lyudmila Valerevna, Dupleva Liliya Shamilevna, Chernov Albert Nikolaevich
Format: Patent
Sprache:eng ; rus
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Zusammenfassung:FIELD: veterinary medicine.SUBSTANCE: invention refers to veterinary microbiology and concerns an enzyme immunoassay test system for serological diagnosis of infectious keratoconjunctivitis in cattle and control of postvaccinal immunity. Disclosed is an immunoenzymometric test system for determining specific antibodies to Moraxella bovoculi bacteria in bovine blood serum, which contains a specific antigen of the patented strain "CX-P6 N-DEP" Moraxella bovoculi, obtained by washing 3 times thereof with 0.01 M phosphate- with buffered saline solution and subsequent destruction of bacterial cells with ultrasound on "Bandelin GM3100" disintegrator at frequency 20 kNz ± 500 Hz for 10 min at 4 °C, further precipitation of Moraxella bovoculi antigen with trichloroacetic acid at 5 % saturation and dialysis against tap water, which is a specific protein in concentration of 9-12 mcg/cm, adsorbed on surface of polystyrene holes in carbonate-bicarbonate buffer. Test system also includes control positive serum prepared for Moraxella bovoculi antigen with antibody titres of 1:6400-1:12800 in ELISA, control negative serum, peroxidase conjugate against IgG bull, buffer solutions: phosphate-buffer solution pH 7.2-7.4 with Tween-20 (detergent) for dilution of control, analyzed serums and washing panels; phosphate-buffer solution pH 7.2-7.4 for conjugate dilution; a chromogen-substrate solution containing ortho-phenylendiamine and hydrogen peroxide, a stop-reagent and panels for the enzyme immunoassay.EFFECT: test system enables diagnosing infectious keratoconjunctivitis in cattle and determining intensity of post-vaccination immunity with high sensitivity and specificity.1 cl, 2 tbl, 5 ex Изобретение относится к области ветеринарной микробиологии и касается иммуноферментной тест-системы для серологической диагностики инфекционного кератоконъюнктивита крупного рогатого скота и контроля напряженности поствакцинального иммунитета. Раскрыта иммуноферментная тест-система для определения специфических антител к бактериям Moraxella bovoculi в сыворотке крови крупного рогатого скота, которая содержит специфический антиген запатентованного штамма «СХ-Ч6 № - ДЕП» Moraxella bovoculi, полученный путем 3-кратного отмыва их 0,01 М фосфатно-буферным физиологическим раствором и последующим разрушением бактериальных клеток ультразвуком на дезинтеграторе «Bandelin GM3100» при частоте 20 kНz ± 500 Гц в течение 10 мин при 4°С, дальнейшим осаждением антигена Moraxella bovoculi трихлоруксусной к