PCR荧光探针法在检测分泌物沙眼衣原体、淋球菌及解脲支原体中的应用

Application of PCR with fluorescent probe to test for Chlamydia trachomatis,Neisseria gonorrhoeae and Ureaplasma urelyticum in secretion

目的评价PCR荧光探针法用于检测分泌物沙眼衣原体(Chlamydia trachomatis,CT)、淋球菌(Neisseria gonorrhoeae,NG)、解脲支原体(Ureaplasma urealyticum,UU)的适用性。方法热裂解法提取CT、NG、UU分泌物样本中病原微生物DNA,以其作为模板,采用PCR荧光探针法检测样本中CT、NG、UU的阳性率。同时采用乳胶法检测相应样本中CT的阳性率,培养法检测相应样本中NG及UU的阳性率,并将检测结果分别与PCR荧光探针法进行比较。结果 1 209份CT标本中,PCR荧光探针法阳性检出率(9.76%)显著高于乳胶法(2.64%),1 491份NG标本中,PCR荧光探针法阳性检出率(6.64%)显著高于培养法(4.16%),差异均有统计学意义(P<0.05);1 260份UU标本中,PCR荧光探针法阳性检出率(37.6%)略高于培养法法(36.8%),但差异无统计学意义(P>0.05)。结论与培养法和乳胶法比较,荧光探针PCR法对CT和NG的阳性检出率更高,而对UU的阳性检出率与培养法相近,适用于分泌物中CT、NG及UU的检测。

Objective To evaluate the suitability of PCR with fluorescent probe in test for Chlamydia trachomatis（CT）,Neisseria gonorrhoeae（NG） and Ureaplasma urelyticum（UU） in secretion. Methods The DNAs of pathogenic microorganisms in CT,NG and UU in secretions were extracted by thermal lysis and used as the templates for determination of positive rates of CT,NG and UU in samples by PCR with fluorescent probe,of which the results were compared with that of CT determined by latex method and those of NG and UU determined by culture method. Results In 1 209 samples of CT,the positive rate determined by PCR with fluorescent probe（9. 76%） was significantly higher than that by latex method（2. 64%）（P 〈0. 05）. In 1 491 samples of NG,the positive rate determined by PCR with fluorescent probe（6. 64%）was significantly higher than that by cutlure method（4. 16%）（P 〈0. 05）. However,in 1 260 samples of UU,the positive rate determined by PCR with fluorescent probe（37. 6%） was slightly higher than that by culture method（36. 8%）（P 〉0. 05）. Conclusion The positive rates of CT and NG determined by PCR with fluorescent probe were higher than,while that of UU was similar to,those by culture method and latex method,indicating that the PCR with fluorescent probe was suitable for deteminates of CT,NG and UU in secretion.