Mádr P, Benda V, Skardová O
Ustav fyziologie a genetiky hospodárských zvírat CSAV, Praha-Uhrínĕves.
Vet Med (Praha). 1990 Apr;35(4):193-9.
A sensitive 4-layers ELISA test for determination of antibodies against the pathogens of Streptococcus agalactiae in cows' milk was used for diagnosis of mastitis, with the aim to broaden these methods. Antigen was linked on the solid phase in the form of the whole bacteria, and milk was tested, diluted in the ratio of 1:10. Antigen bound-specific antibodies were labelled with pig antibodies against bovine immunoglobulins and in the next layer with rabbit antibody conjugated with peroxidases against pig immunoglobulins. After test visualisation and reading on the photometre, the results were given in the positivity per cent as a 100-multiple of the proportion of absorbance of the unknown sample and the positive control after subtraction of the negative control. Milk was examined in 36 dairy cows from three various breeding herds by that method. The samples were parallelly examined bacteriologically and cytologically. In the milk of dairy cows with positive S. agalactiae finding, the main level of antibodies expressed a positivity per cent, was 15.0%, while in bacteriologically negative animals it was only 6.2%. The dairy cows were divided into 8 groups, characterizing various stages of mastitis, according to the results of the individual treatments.
采用一种灵敏的四层酶联免疫吸附试验（ELISA）检测牛奶中抗无乳链球菌病原体的抗体，以诊断乳腺炎，目的是拓宽这些方法。抗原以全菌形式连接在固相上，牛奶按1:10的比例稀释后进行检测。抗原结合的特异性抗体用抗牛免疫球蛋白的猪抗体标记，在接下来的一层中用与抗猪免疫球蛋白结合的过氧化物酶的兔抗体标记。在光度计上进行检测可视化和读数后，结果以阳性百分比给出，即未知样品与阳性对照的吸光度比例减去阴性对照后乘以100。用该方法对来自三个不同育种牛群的36头奶牛的牛奶进行了检测。对样品同时进行了细菌学和细胞学检测。在无乳链球菌检测呈阳性的奶牛的牛奶中，抗体的主要水平以阳性百分比表示为15.0%，而在细菌学检测呈阴性的动物中仅为6.2%。根据个体治疗结果，将奶牛分为8组，以表征乳腺炎的不同阶段。
