应用连接酶链反应检测沙眼衣原体DNA

目的 建立检测沙眼农原体感染的具有高度敏感性和特异性的缺口—连接酶链反应—酶联免疫吸附测定法。方法 根据CT主要外膜蛋白稳定区设计2对互补探针并分别对其两端标记生物素和地高辛，对6种CT标准株、鹦鹉热衣原体标准株和其他细菌进行Gap—LCR反应并以ELISA和EB染色电泳检测扩增产物以比较两者的检测限度。结果Gap LCR可检出6种CT标准株并不与其他细菌发生交叉反应，ELISA可检出10fg DNA模板的扩增产物，较EB电泳法敏感10倍。结论 Gap—LCR—ELISA是一高度敏感特异的检测CT感染的核酸扩增技术，是适宜于我国广大医疗基层单位进行大规模CT分子流行病学研究的极具前景的方法。

Gap ligase chain reaction enzyme linked immunoadsorbent assay (Gap-LCR-ELISA) for diagnosis of chlamydia trachomatis

Objective To establish a sensitive,specific and reliable Gap LCR ELISA for diagnosis of chlamydia trachomatis (CT).Methods A serial of Gap LCR DNA amplification assays were conducted to 6 strains of CT,2 strains of C psittaci and some other bacteria.The 2 pairs of probes labeled with biotin and digoxinine were derived from the CT outer major membrane protein gene (omp1).The amplification products were detected by ELISA method and PAGE to compare their detection limits.Results Gap LCR using labled probes could detect six strains of CT and had no cross reactions with other bacteria.The detection limit of ELISA was 10fg template DNA which was 10 fold sensitive than PAGE to detect the amplified products.Conclusion Gap LCR ELISA is a highly sensitive and specific nucleic acid amplification technique to diagnose chlamydial infections,which is a promising method for large scale CT molecular epidemic investigation in our country.