Deguchi T, Yamamoto H, Tada K, Komeda H, Iwata H, Ito Y, Saito A, Ban Y, Tamaki M, Maeda S
Department of Urology, Gifu University School of Medicine.
Kansenshogaku Zasshi. 1992 May;66(5):555-60. doi: 10.11150/kansenshogakuzasshi1970.66.555.
A polymerase chain reaction (PCR) procedure was developed for detection of Neisseria gonorrhoeae. Two oligonucleotides based on sequences within a 16S ribosomal RNA gene from N. gonorrhoeae were used as extension primers for the PCR. A single DNA fragment of 206 bp was amplified, when N. gonorrhoeae DNA was template for the PCR. No amplified product was detected in Chlamydia trachomatis DNA, Ureaplasma urealyticum DNA or other bacterial DNAs. The DNA fragment of 206 bp was detected on agarose gel electrophoresis, when DNA of greater than or equal to 6.5 N. gonorrhoeae per PCR was used as template DNA for the PCR. The culture and the PCR were carried out for detection of N. gonorrhoeae in 67 urethral swabs obtained from male patients with urethritis. In 27 of 28 specimens in which N. gonorrhoeae was isolated and identified by the culture, 206 bp DNA fragment was amplified by the PCR, but in one specimen no DNA fragment was detected. In 2 of 39 culture-negative specimens, 206 pb DNA fragment was detected and in the remaining specimens, PCR was negative for N. gonorrhoeae. The overall detection coincidence rate between the culture and the PCR was 95.5% (64/67). Thus, the PCR procedure developed in this study was sensitive and specific for detection of N. gonorrhoeae and could be applied for diagnosis of gonococcal urethritis.
开发了一种用于检测淋病奈瑟菌的聚合酶链反应（PCR）程序。基于淋病奈瑟菌16S核糖体RNA基因内序列的两种寡核苷酸用作PCR的延伸引物。以淋病奈瑟菌DNA为PCR模板时，可扩增出一条206bp的单一DNA片段。在沙眼衣原体DNA、解脲脲原体DNA或其他细菌DNA中未检测到扩增产物。当每PCR使用大于或等于6.5个淋病奈瑟菌的DNA作为PCR模板DNA时，在琼脂糖凝胶电泳上可检测到206bp的DNA片段。对67例男性尿道炎患者的尿道拭子进行淋病奈瑟菌培养和PCR检测。在28份经培养分离并鉴定出淋病奈瑟菌的标本中，有27份通过PCR扩增出206bp的DNA片段，但有1份标本未检测到DNA片段。在39份培养阴性的标本中，有2份检测到206pb的DNA片段，其余标本淋病奈瑟菌PCR检测为阴性。培养与PCR的总体检测符合率为95.5%（64/67）。因此，本研究开发的PCR程序对淋病奈瑟菌的检测具有敏感性和特异性，可用于淋菌性尿道炎的诊断。
