Gu Linlin, Wang Weiwei, Yang Mingjie, Yu Yafang, Chen Hui, Zhang Kang, Yuan Kehu, Fu Hui, Mo Qiuhua, Yang Bo
Research and Development Department, Shenzhen Bioeasy Biotechnology Co., Ltd, Shenzhen, Guangdong, 518101, China.
Lanzhou Institute of Husbandry and Pharmaceutical Sciences, Chinese Academy of Agricultural Sciences, Lanzhou, 730050, China.
Appl Microbiol Biotechnol. 2025 Jul 1;109(1):155. doi: 10.1007/s00253-025-13542-x.
Early pregnancy diagnosis in dairy cows is an important part of the production processes. Accurate early pregnancy diagnosis can help veterinarians identify open cow as soon as possible and carry out the second breeding earlier, which is of great significance for improving reproductive efficiency of dairy cow. In this study, a rapid immunochromatography based on latex microspheres for detecting pregnancy-related glycoprotein (PAG) was established with two monoclonal antibodies. The samples collected from 28 days after breeding of 559 Holstein cows were used to diagnose pregnancy. The results showed that the minimum detection limit of the established latex immunochromatography for PAG was 0.2 ng/mL. Compared to ultrasound, the sensitivity, specificity, positive predictive value, negative predictive value, and accuracy of this method for pregnancy diagnosis was 99.25%, 90%, 91.03%, 99.15%, and 94.68%, respectively, and the kappa value is 0.89, which demonstrated that the pregnancy diagnosis results of the test strip method have high consistency with ultrasound. Compared with commercial enzyme-linked immunosorbent assay (ELISA) kits, the positive coincidence rate, negative coincidence rate, and overall coincidence rate of this method was 98.29%, 97.6%, and 98.03%, respectively. This indicated that the test strip has high consistency with the currently widely used ELISA immunological diagnostic method. The latex immunochromatographic method established in this study can accurately diagnose pregnant cows and can be used for early pregnancy diagnosis of dairy cows. The method is simple to operate and fast to detect, and the results are easy to interpret, which is of great significance and clinical application value for cow early pregnancy detection. KEY POINTS: • A rapid immunochromatography based on latex microspheres with two monoclonal antibodies specific to PAG2 was established for early pregnancy diagnosis of dairy cows. • The early pregnancy test method, which we have developed, exhibits high sensitivity and can be utilized in small to medium-scale ranches. • The clinical sample test results demonstrated that the test strip has a high degree of consistency with ultrasound.
奶牛早期妊娠诊断是生产过程中的重要环节。准确的早期妊娠诊断有助于兽医尽快识别空怀母牛并尽早进行第二次配种，这对提高奶牛繁殖效率具有重要意义。本研究利用两种单克隆抗体建立了一种基于乳胶微球的快速免疫层析法检测妊娠相关糖蛋白（PAG）。收集559头荷斯坦奶牛配种后28天的样本用于妊娠诊断。结果表明，所建立的乳胶免疫层析法对PAG的最低检测限为0.2 ng/mL。与超声检查相比，该方法用于妊娠诊断的灵敏度、特异度、阳性预测值、阴性预测值和准确度分别为99.25%、90%、91.03%、99.15%和94.68%，kappa值为0.89，表明试纸条法的妊娠诊断结果与超声检查具有高度一致性。与市售酶联免疫吸附测定（ELISA）试剂盒相比，该方法的阳性符合率、阴性符合率和总符合率分别为98.29%、97.6%和98.03%。这表明试纸条与目前广泛使用的ELISA免疫诊断方法具有高度一致性。本研究建立的乳胶免疫层析法能够准确诊断妊娠母牛，可用于奶牛早期妊娠诊断。该方法操作简便、检测快速，结果易于判读，对奶牛早期妊娠检测具有重要意义和临床应用价值。要点：• 建立了一种基于乳胶微球的快速免疫层析法，使用两种对PAG2特异的单克隆抗体用于奶牛早期妊娠诊断。• 我们开发的早期妊娠检测方法具有高灵敏度，可用于中小型牧场。• 临床样本检测结果表明试纸条与超声检查具有高度一致性。
