Chui L, Chiu T, Kakulphimp J, Tyrrell G J
Department of Laboratory Medicine and Pathology, University of Alberta, Edmonton, AB, Canada.
Clin Microbiol Infect. 2008 May;14(5):473-9. doi: 10.1111/j.1469-0691.2008.01950.x. Epub 2008 Feb 22.
Three different real-time PCR assays were evaluated as confirmatory tests for Neisseria gonorrhoeae after initial screening using the COBAS AMPLICOR Chlamydia trachomatis and N. gonorrhoeae duplex assay. The target genes used for the confirmation were the gyr, cppB and 16S rRNA genes. Analytical specificity was determined by testing 60 strains belonging to different bacterial species and/or serogroups. The primers chosen from the 16S rRNA gene for confirmation of N. gonorrhoeae were highly specific, showed no cross-reactivity with other bacteria included in the study, and had an analytical sensitivity of 1 CFU. Of 192 clinical specimens that were positive for N. gonorrhoeae according to the COBAS AMPLICOR assay, 42 were confirmed as positive using the 16S rRNA gene target, 26 were confirmed using the cppB target, and 30 were confirmed using the gyr target. It was concluded that the real-time PCR assay targeting the 16S rRNA gene is a useful confirmatory assay to complement the COBAS AMPLICOR screening test for N. gonorrhoeae.
在使用COBAS AMPLICOR沙眼衣原体和淋病奈瑟菌双重检测法进行初步筛查后，对三种不同的实时荧光定量聚合酶链反应（PCR）检测方法进行了评估，作为淋病奈瑟菌的确证试验。用于确证的靶基因是gyr、cppB和16S rRNA基因。通过检测属于不同细菌物种和/或血清群的60株菌株来确定分析特异性。从16S rRNA基因中选择用于确证淋病奈瑟菌的引物具有高度特异性，与研究中包含的其他细菌无交叉反应，分析灵敏度为1 CFU。在COBAS AMPLICOR检测法检测为淋病奈瑟菌阳性的192份临床标本中，42份使用16S rRNA基因靶标确证为阳性，26份使用cppB靶标确证为阳性，30份使用gyr靶标确证为阳性。得出结论，针对16S rRNA基因的实时荧光定量PCR检测法是一种有用的确证检测法，可补充COBAS AMPLICOR淋病奈瑟菌筛查试验。
