肺炎衣原体套式聚合酶链式反应检测研究

摘要

Aim To develop a molecular biologic technique for detection of chlamudia pneumoniae with nested polymeyase chain reaction (nPCR) .Methods Nested primers were synthesized according to a cloned C.pneumoniae 474- bp Pst I fragmert. Results The 378bp DNA fragments were amplified from C. pneuomoniae with nPCR. None of the C. trachomatis, C. psittaci, other organisms and etc. Strains tested were amplified by the nPCR. The direct sequening of 3 sample products with nPCR are quite same as C. pneumoniae (CWL-29 ) . The sensitivity of nPCR is higter than PCR. Conclusions This method is not only sensitive.specific and rapid, but also provides an etiological basis for cdiagnosis of C.pneumoniae infection.%目的　探讨肺炎衣原体(Cpn)的套式聚合酶链式反应(nPCR)检测技术。方法　根据Cambell克隆的CpnDNA序列设计套式引物，并建立nPCR反应体系。结果　 Cpn经nPCR扩增出现378bp的阳性条带，而沙眼衣原体和鹦鹉热衣原体以及其他用作特异性试验的微生物均不能扩增。3例阳性标本经DNA测序与CWL(CW-29株)序列完全一致。CPN nPCR之灵敏度比传统PCR高100倍。各种呼吸系统病患儿咽拭子Cpn nPCR阳性率明显高于健康儿童(P均<0.01)。结论　 nPCR检测可能是Cpn感染的灵敏、特异、快速的诊断方法　。