涂片与培养及PCR/RLB检测淋病奈瑟菌的比较研究

The study of Comparing with Smear, Culture and PCR-reverse Line Hybridization Assay for Detecting Neisseria Gonorrhoeae

目的:建立以16SrRNA为靶基因的PCR-反向线点杂交技术(RLB)检测淋病奈瑟菌(Neisseria gonor-rhoeae,NG),并与涂片法及培养法比较。方法:选择NG16SrRNA基因设计一对PCR引物,生物素标记下游引物扩增NGDNA,然后与固定在尼龙膜上的特异性寡核苷核探针杂交。并对115例性病高危人群标本进行检测,然后与涂片法与培养法检测结果进行比较。结果:PCR扩增NGDNA的片段长度分别为414bp。通过对115例临床标本的检测,PCR/RLB的阳性率为31.3%,而涂片法和培养法的阳性率分别为15.7%和21.7%。结论:PCR/RLB是一种快速、敏感的检测方法,对NG感染的实验诊断具有十分重要的意义。

Objective To develop 16S rRNA PCR-reverse line blot hybridization （RLB） assay for detection of N. gonorrhoeae and compare it with smear and culture methods. Methods The DNA from N. gonorrhoeae was amplified with the one set of primer for the 16S rRNA gene, The reverse primers were labeled with biotin. The products were identified by hybridization with its specific oligonueleotide probes labeled with amidogen in a nylon membrane, Then using PCR-RLB to detect 115 specimens and compare it with smear and culture methods. Results The length of PCR products was 414 bp. The PCR-RLB positive rate （31.3 % ） is higher than smear （ 15.7% ） and culture （21.7% ） （ P 〈 0.05 ）. Conclusion The results of PCRRLB in detecting N. gonorrhoeae also provided excellent results. It plays the very important role in detecting of clinical pathogens.