实验动物科学

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实验大鼠肺支原体ELISA检测方法的建立

贺爭鸣;刘佐民;王春玲;孙光;吴惠英;卫礼;张然;   

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  • 收稿日期:1992-09-25 修回日期:1992-09-25 出版日期:1992-09-25 发布日期:1992-09-25

Establishment of ELISA for the Detection of Antibodies to Mycoplasma pulmonis in Rats

He Zhengming Liu Zuomin Wang Chunling Sun Guang Wu Huiying Wei Li Zhang Ran National Institute for the Control of Pharmacentical and Biological Products Bcijing, 100050   

  • Received:1992-09-25 Revised:1992-09-25 Online:1992-09-25 Published:1992-09-25

摘要: 报告了实验大鼠肺支原体感染ELISA检测方法的研究。抗原包被量在0.5μg/ml,酶结合物1:16000稀释时,P/N值最大。波长在492时O.D值随抗原和抗体浓度不同而呈现有规律的变化。阳性血清稀释至1:1280时,P/N值仍在阳性范围之内。与常规的分离培养方法相比较,ELISA的阳性检出率高,且阳性血清的ELISA阻断率均大于50%。重复测定,实验结果的再现性好。以上结果说明所建立的ELISA方法,其敏感性、特异性和重复性均好。在所检测的130只普通级大鼠中,抗体阳性率达91%。另外还发现在实验大鼠中还存在关节支原体的感染,且一般与肺支原体以混合感染的形式存在。在清洁级大鼠中未见这两种支原体的感染。

关键词: 肺支原体, ELISA

Abstract: This paper reported the reseazch work on the establishment of ELISA used for the detection of antibodies against M pulmonis in rats. The reaction with 5μg of lysate antigen per ml and conjugate dilution of 1:16000 resulted in maximum ratio of P/N. OD_(492) value was varied with concentration of antigen and antibodies regularly. The ratio of P/N was higher than 2.1, even though serum was diluted in 1: 1280. Compared with the cultural isolation, ELISA is more sensitive in the detection of natural infection of M. pulmonis and the blocking ratio of positive sera was higher then 50%. Evidence above suggested that sensitivity, specificity, and reproducibility of ELISA system are satisfactory. M. pulmonis infcetion was found in 91% of 130 conventional rats tested. The survey also showed the infection of M. arthritidis, Which is mixed infection with M pulmonis. No infection of these two mycoplasma was found in clear rats.

Key words: M. pulmonis, ELISA