McLaughlin G L, Howe D K, Biggs D R, Smith A R, Ludwinski P, Fox B C, Tripathy D N, Frasch C E, Wenger J D, Carey R B
Department of Veterinary Pathobiology, School of Veterinary Medicine, Purdue University, West Lafayette, IN 47907.
Mol Cell Probes. 1993 Feb;7(1):7-17. doi: 10.1006/mcpr.1993.1002.
In 1991-92, Neisseria meningitidis group C was isolated from the blood of eight students in Urbana, Illinois, USA, and from the cerebrospinal fluid of one student from a nearby community, Decatur, Illinois. These and other bacterial species were analysed by PCR fingerprinting using primers selected from the ribosomal (r)DNA loci. A rDNA primer pair spanning a region within the 16S rDNA amplified a predicted 280 base pair (bp) DNA fragment from Neisseria spp. and fragments of different sizes for other genera. This primer pair specifically detected a carrier of N. meningitidis in a small clinical battery. Identity of the fragment was confirmed by restriction endonuclease analysis. A 600 bp fragment was also amplified from the 16S-23S internal transcribed spacer (ITS) of N. meningitidis; amplification from six other genera yielded different-sized fragments. Digestion of the ITS fragment from N. meningitidis with Alu I revealed three patterns; pattern I was found only for serogroup C isolates, and it was the dominant pattern among recent isolates with the exception of the one from Decatur. The isolate from Decatur yielded pattern III which suggested a non-clonal relationship to the seven isolates from Urbana. Patterns II and III were more prevalent in isolates from the 1960's and 1980's. PCR-based analysis of these loci can complement the techniques which are currently used for the detection and typing of these and other eubacteria.
1991 - 1992年，在美国伊利诺伊州厄巴纳市，从8名学生的血液中分离出了C群脑膜炎奈瑟菌，在伊利诺伊州迪凯特市附近社区的1名学生的脑脊液中也分离出了该菌。使用从核糖体（r）DNA位点选择的引物，通过PCR指纹分析对这些细菌及其他细菌种类进行了分析。一对跨越16S rDNA内一个区域的rDNA引物，从奈瑟菌属扩增出了一个预测的280碱基对（bp）的DNA片段，而从其他属扩增出了不同大小的片段。这对引物在一个小型临床样本中特异性地检测出了脑膜炎奈瑟菌的携带者。通过限制性内切酶分析确认了该片段的同一性。还从脑膜炎奈瑟菌的16S - 23S内部转录间隔区（ITS）扩增出了一个600 bp的片段；从其他六个属扩增出的片段大小不同。用Alu I消化脑膜炎奈瑟菌的ITS片段显示出三种模式；模式I仅在C群血清型分离株中发现，并且是近期分离株中的主要模式，但迪凯特市的分离株除外。迪凯特市的分离株产生模式III，这表明它与厄巴纳市的七个分离株没有克隆关系。模式II和III在20世纪60年代和80年代的分离株中更为普遍。基于PCR对这些位点的分析可以补充目前用于检测和分型这些及其他真细菌的技术。
