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单克隆抗体在山羊传染性胸膜肺炎(CCPP)诊断中的应用。

The use of monoclonal antibodies in the diagnosis of contagious caprine pleuropneumonia (CCPP).

作者信息

Thiaucourt F, Bölske G, Libeau G, Le Goff C, Lefèvre P C

机构信息

PATHOTROP CIRAD-EMVT, Maisons Alfort, France.

出版信息

Vet Microbiol. 1994 Aug 1;41(3):191-203. doi: 10.1016/0378-1135(94)90100-7.

DOI:10.1016/0378-1135(94)90100-7
PMID:7975145
Abstract

Contagious caprine pleuropneumonia is a severe disease affecting goats in Eastern Africa and the Middle East, caused by Mycoplasma sp. type F38. Its exact geographical distribution is however not exactly known due to the lack of specificity of the available serological tests and the difficulty in cultivating M. sp. F38. A panel of monoclonal antibodies (mAbs) was produced, using crude or membrane proteins antigens from type F38 strains to immunize mice. The reactivity of the mAbs was tested by an immunobinding assay with crude mycoplasma antigens spotted on nitrocellulose filters. One hundred and twelve antigens, standardized at 0.5 mg protein/ml, were used. Mycoplasma strains were chosen among closely related species of the "mycoides cluster", M. capricolum, Group 7 of Leach, M. mycoides mycoides LC, M. mycoides mycoides SC, M. mycoides capri, as well as among species that are isolated from goat lungs, M. arginini, M. ovipneumoniae, M. putrefaciens, M. agalactiae. Out of 60 mAbs, 4 were chosen to build an identification test for mycoplasmas of the "mycoides cluster". Controls showed that accurate identification could be hampered by antigenic heterogeneity within the M. capricolum species. One mAb was used for the direct detection of M. sp. F38 antigen in pleural fluid from goats suspected of CCPP. The sensitivity of the test can be estimated at 0.5 micrograms protein/ml. Comparison with isolation results show a 74% agreement between the two methods. The same mAb was used to build a blocking ELISA. This serological test was strictly specific for CCPP. It detects antibodies in sera of naturally infected or artificially immunized animals while it remained negative with hyperimmune sera to related strains such as PG 50. Direct antigen detection and blocking ELISA are tools that may enable a better assessment of CCPP distribution.

摘要

山羊传染性胸膜肺炎是一种影响东非和中东地区山羊的严重疾病,由F38型支原体引起。然而,由于现有血清学检测缺乏特异性以及F38型支原体培养困难,其确切地理分布尚不完全清楚。利用F38型菌株的粗提物或膜蛋白抗原免疫小鼠,制备了一组单克隆抗体(mAb)。通过免疫结合试验检测mAb与点样于硝酸纤维素滤膜上的支原体粗抗原的反应性。使用了112种标准化为0.5mg蛋白质/ml的抗原。从“丝状支原体簇”的近缘物种中选择支原体菌株,包括山羊支原体、利奇7组、丝状支原体丝状亚种LC、丝状支原体丝状亚种SC、丝状支原体山羊亚种,以及从山羊肺中分离出的物种,如精氨酸支原体、绵羊肺炎支原体、腐败支原体、无乳支原体。在60种mAb中,选择了4种构建“丝状支原体簇”支原体的鉴定试验。对照显示,山羊支原体物种内的抗原异质性可能会妨碍准确鉴定。一种mAb用于直接检测疑似患有山羊传染性胸膜肺炎的山羊胸腔积液中的F38型支原体抗原。该试验的灵敏度估计为0.5微克蛋白质/ml。与分离结果比较显示,两种方法的一致性为74%。使用相同的mAb构建了阻断ELISA。这种血清学检测对山羊传染性胸膜肺炎具有严格的特异性。它能检测自然感染或人工免疫动物血清中的抗体,而对PG 50等相关菌株的超免疫血清仍呈阴性。直接抗原检测和阻断ELISA可能有助于更好地评估山羊传染性胸膜肺炎的分布情况。