Gen-Probe Incorporated, San Diego, California, USA.
J Clin Microbiol. 2013 Oct;51(10):3199-206. doi: 10.1128/JCM.00309-13. Epub 2013 Jul 17.
An apparently rare Neisseria meningitidis isolate containing one copy of a Neisseria gonorrhoeae 16S rRNA gene is described herein. This isolate was identified as N. meningitidis by biochemical identification methods but generated a positive signal with Gen-Probe Aptima assays for the detection of Neisseria gonorrhoeae. Direct 16S rRNA gene sequencing of the purified isolate revealed mixed bases in signature regions that allow for discrimination between N. meningitidis and N. gonorrhoeae. The mixed bases were resolved by sequencing individually PCR-amplified single copies of the genomic 16S rRNA gene. A total of 121 discrete sequences were obtained; 92 (76%) were N. meningitidis sequences, and 29 (24%) were N. gonorrhoeae sequences. Based on the ratio of species-specific sequences, the N. meningitidis strain seems to have replaced one of its four intrinsic 16S rRNA genes with the gonococcal gene. Fluorescence in situ hybridization (FISH) probes specific for meningococcal and gonococcal rRNA were used to demonstrate the expression of the rRNA genes. Interestingly, the clinical isolate described here expresses both N. meningitidis and N. gonorrhoeae 16S rRNA genes, as shown by positive FISH signals with both probes. This explains why the probes for N. gonorrhoeae in the Gen-Probe Aptima assays cross-react with this N. meningitidis isolate. The N. meningitidis isolate described must have obtained N. gonorrhoeae-specific DNA through interspecies recombination.
本文描述了一株含有一个淋病奈瑟菌 16S rRNA 基因拷贝的脑膜炎奈瑟菌分离株。该分离株通过生化鉴定方法被鉴定为脑膜炎奈瑟菌,但在用于检测淋病奈瑟菌的 Gen-Probe Aptima 检测中产生了阳性信号。对纯化分离株的 16S rRNA 基因进行直接测序,发现特征区域存在混合碱基,可区分脑膜炎奈瑟菌和淋病奈瑟菌。通过单独测序扩增的基因组 16S rRNA 基因的单个拷贝来解决混合碱基问题。总共获得了 121 个离散序列;92 个(76%)是脑膜炎奈瑟菌序列,29 个(24%)是淋病奈瑟菌序列。基于种特异性序列的比例,该脑膜炎奈瑟菌菌株似乎用淋球菌基因替换了其四个固有 16S rRNA 基因之一。使用针对脑膜炎球菌和淋病球菌 rRNA 的荧光原位杂交(FISH)探针来证明 rRNA 基因的表达。有趣的是,这里描述的临床分离株表达脑膜炎奈瑟菌和淋病奈瑟菌 16S rRNA 基因,如两种探针的阳性 FISH 信号所示。这解释了为什么 Gen-Probe Aptima 检测中的淋病奈瑟菌探针与该脑膜炎奈瑟菌分离株发生交叉反应。描述的脑膜炎奈瑟菌分离株必须通过种间重组获得淋病奈瑟菌特异性 DNA。