Molina Caballero J M, Anguiano A, Ferrer O, Serrano E, Uceda A
Departamento de Patología Animal, Facultad de Veterinaria, Universidad de Las Palmas de Gran Canaria, Spain.
Rev Sci Tech. 1993 Jun;12(2):629-38. doi: 10.20506/rst.12.2.712.
An enzyme-linked immunosorbent assay (ELISA) was performed for diagnosis of paratuberculosis in goats, using as antigen a protoplasmic extract (PPA-3). The test was developed on the basis of the results obtained with two serum reference pools, positive and negative respectively. To avoid day-to-day variations, dilutions of the positive serum pool were included in each plate to obtain an arbitrary system, transforming absorbance into immunoglobulin (Ig) G anti-Mycobacterium paratuberculosis units. The ELISA was used on sera of two reference groups of animals. One group consisted of 35 goats suspected of being infected with paratuberculosis, which was confirmed by histological findings and isolation of M. paratuberculosis. The negative group consisted of 61 healthy goats from a farm free of paratuberculosis. The test showed a sensitivity of 100% and a specificity of 91.8%. Absorption of sera with a Mycobacterium phlei suspension did not modify either the sensitivity or the specificity of the test. Sera from the negative group were analysed by Western blotting, and four of them recognized two fractions with a molecular weight of 17.3 and 28.1 kDa.
采用酶联免疫吸附测定（ELISA）法，以原生质提取物（PPA - 3）作为抗原诊断山羊副结核病。该检测方法是根据分别用两个血清参考库（阳性和阴性）所获结果开发的。为避免日常差异，每块板中均加入阳性血清库的稀释液，以获得一个任意系统，将吸光度转化为抗副结核分枝杆菌免疫球蛋白（Ig）G单位。ELISA法用于两组动物的血清检测。一组由35只疑似感染副结核病的山羊组成，经组织学检查结果和副结核分枝杆菌分离鉴定得到确诊。阴性组由来自一个无副结核病农场的61只健康山羊组成。该检测显示敏感性为100%，特异性为91.8%。用草分枝杆菌悬液吸收血清，既不改变检测的敏感性，也不改变其特异性。对阴性组的血清进行免疫印迹分析，其中四份血清识别出分子量为17.3 kDa和28.1 kDa的两个条带。
