RUDN Journal of Agronomy and Animal IndustriesRUDN Journal of Agronomy and Animal IndustriesВестник Российского университета дружбы народов. Серия: Агрономия и животноводство2312-797X2312-7988Peoples’ Friendship University of Russia named after Patrice Lumumba1966510.22363/2312-797X-2021-16-2-154-166Animal breedingЖивотноводствоResearch ArticleUsing commercial enzyme immunoassay for measuring pregnancy-associated glycoproteins to diagnose pregnancy in dairy cows under field conditions in AlgeriaИспользование иммуноферментного анализа для определения гликопротеинов беременности с целью диагностики стельности дойных коров в полевых условиях Алжираhttps://orcid.org/0000-0002-9325-7889AyadAbdelhanineАядАбдельханин

DVM, MSc, PhD, Department of Biological Sciences of the Environment, Faculty of Nature and Life Sciences

доктор медицинских наук, кафедра общей экологии, факультет естественных наук

abdelhanine.ayad@univ-bejaia.dzhttps://orcid.org/0000-0003-0459-8399DerbakHananeДербакХанане

MSc, PhD Student, Department of Biological Sciences of the Environment, Faculty of Nature and Life Sciences

аспирант, кафедра общей экологии, факультет естественных наук

derbak90@hotmail.comhttps://orcid.org/0000-0003-1365-4479BessebouaOmarБессебуаОмар

DVM, MSc, PhD, Department of Agronomic and Biotechnological Sciences, Faculty of Nature and Life Sciences

доктор медицинских наук, кафедра агрономии и биотехнологии, факультет естественных наук

besseboua.omar@gmail.comUniversity of BejaiaУниверситет Абдеррахмана МираHassiba Benbouali University of ChlefУниверситет Хассибы Бенбуали07072021162VOL 16, NO2 (2021)ТОМ 16, №2 (2021)15416607072021Copyright ©; 2021, Ayad A., Derbak H., Besseboua O.Copyright ©; 2021, Аяд А., Дербак Х., Бессебуа О.2021Ayad A., Derbak H., Besseboua O.Аяд А., Дербак Х., Бессебуа О.http://creativecommons.org/licenses/by/4.0https://agrojournal.rudn.ru/agronomy/article/view/19665

The purpose of the present work was to study effectiveness for early pregnancy diagnosis in cattle of the new enzyme immunoassay (EIA) sandwich kit commercially available based on the measurement of pregnancy-associated glycoproteins (PAGs). 120 Holstein-Friesian cattle of mixed age and parity were comprised from different dairy herds. The pregnant females (n = 68) were diagnosed by ultrasonography at day 35-40 after artificial insemination and confirmed by transrectal exploration at 2-3 months after AI. The non-pregnant females (n = 52) were housed in the absence of males during the experimental period. Blood samples were collected from coccygeal vessels of females into EDTA tubes. The serum was obtained by centrifugation and the serum was stored at - 20 °C until assay. The PAG concentrations in pregnant and non-pregnant females were determined in serum by EIA kit. The reproducibility inter- and intra-assay of the PAG-EIA is satisfactory (2.78 and 13.19 %, respectively). The accuracy (≥ 94.8 %) and the test of parallelism were largely acceptable. No cross-reaction was observed with the different hormones tested at different dilutions. PAG-EIA system gave 100 % sensitivity and negative predictive values. Whereas, specificity and positive predictive value were 91.93 and 71.15 %, respectively. The accuracy of pregnancy diagnosis by PAG-EIA was 87.5 %. In conclusion, the present study shows clearly that the EIA kit can be used to measure PAG in serum cows for the detection of gestation in Algeria. Therefore, this alternative technique could be recommended to replace the radioactive methods in immunoassays to improve the reproductive performances and an efficient tool for reproductive management of dairy cattle.

Изучена эффективность ранней диагностики беременности у крупного рогатого скота в полевых условиях в Алжире с помощью нового доступного набора для иммуноферментного анализа, основанного на определении гликопротеинов, связанных с беременностью. Исследованы 120 коров голштино-фризской породы смешанного возраста из разных молочных стад. Контроль беременности у 68 стельных коров осуществляли методом ультразвуковой диагностики на 35…40-й день после инсеминации и подтверждали трансректальным исследованием через 2-3 месяца после проведения искусственного осеменения. 52 нестельных коров содержали изолированно от самцов на протяжении всего эксперимента. Образцы крови отбирались из хвостовой артерии в пробирки с ЭДТА. Плазму получали центрифугированием и хранили при температуре -20 °C до начала проведения анализа. Концентрации гликопротеинов беременности (ГПБ) у стельных и нестельных коров определяли в плазме крови с помощью набора реагентов ИФА. Воспроизводимость результатов ГПБ-ИФА (внутри- и межсерийной сходимости) удовлетворительна (2,78 и 13,19 % соответственно). Точность (≥ 94,8 %) и повторяемость анализа были в основном приемлемыми. Не наблюдалось перекрестной реакции с разными гормонами, протестированными в разных разведениях. Диагностическая чувствительность ГПБ-ИФА тест-системы составила 100 % и наблюдалась отрицательная прогностическая значимость. При этом специфичность и прогностическая ценность положительного результата составили 91,93 и 71,15 % соответственно. Точность диагностики беременности с помощью ГПБ-ИФА составила 87,5 %. Таким образом, настоящее исследование явно показывает, что набор для ИФА можно использовать для измерения содержания ГПБ в сыворотке крови коров с целью определения стельности. Следовательно, данный альтернативный метод может быть рекомендован для замены методов иммуноанализа с применением радиоактивных изотопов для улучшения репродуктивных показателей коров и является эффективным инструментом для воспроизводства молочного крупного рогатого скота.

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