Linton D, Dewhirst F E, Clewley J P, Owen R J, Burnens A P, Stanley J
National Collection of Type Cultures, Central Public Health Laboratory, London, UK.
Microbiology (Reading). 1994 Apr;140 ( Pt 4):847-55. doi: 10.1099/00221287-140-4-847.
In the recently described species Campylobacter helveticus, two sizes of PCR amplicon were detected with primers homologous to conserved regions of the 16S rRNA gene. A conventionally sized gene was sequenced from the type strain, NCTC 12470, placing the new species as phylogenetically related to C. upsaliensis and the thermotolerant campylobacters. This nucleotide sequence enabled PCR primers to be designed for use in rapid molecular identification of C. helveticus and its closest phylogenetic relative, C. upsaliensis. When this assay was employed to characterize 22 'C. upsaliensis-like' isolates, twelve were identified as C. helveticus and nine as C. upsaliensis, in agreement with data obtained with a C. helveticus-specific DNA probe. A 550 bp amplicon internal to the 16S rRNA gene of C. helveticus was used to determine restriction fragment length polymorphisms (RFLPs) in genomic Southern blots, confirming that the copy number of the C. helveticus gene was three, and identifying nine 16S rRNA gene profiles. In 5/12 C. helveticus isolates identified by PCR, an enlarged amplicon was detected. The enlarged 16S rRNA gene of one of these strains, NCTC 12838, was sequenced and shown to contain an atypical intervening sequence (IVS) of 148 nucleotides. The position and size of such an IVS was inferred in the other four isolates by PCR with primers 5' and 3' to its position in NCTC 12838. This is a first report of an IVS in the 16S rRNA gene of a eubacterium.
在最近描述的瑞士弯曲杆菌(Campylobacter helveticus)物种中,使用与16S rRNA基因保守区域同源的引物检测到两种大小的PCR扩增子。从模式菌株NCTC 12470中对常规大小的基因进行了测序,将该新物种在系统发育上定位为与乌普萨拉弯曲杆菌(C. upsaliensis)和耐热弯曲杆菌相关。该核苷酸序列使得能够设计用于快速分子鉴定瑞士弯曲杆菌及其最密切的系统发育相关物种——乌普萨拉弯曲杆菌的PCR引物。当采用该检测方法对22株“类乌普萨拉弯曲杆菌”分离株进行鉴定时,12株被鉴定为瑞士弯曲杆菌,9株被鉴定为乌普萨拉弯曲杆菌,这与使用瑞士弯曲杆菌特异性DNA探针获得的数据一致。利用瑞士弯曲杆菌16S rRNA基因内部的一个550 bp扩增子来确定基因组Southern杂交中的限制性片段长度多态性(RFLP),证实瑞士弯曲杆菌基因的拷贝数为3,并鉴定出9种16S rRNA基因图谱。在通过PCR鉴定的12株瑞士弯曲杆菌分离株中,有5株检测到扩增子增大。对其中一株菌株NCTC 12838的增大的16S rRNA基因进行了测序,结果显示其含有一个148个核苷酸的非典型间隔序列(IVS)。通过使用位于NCTC 12838中该IVS位置5'和3'端的引物进行PCR,推断出其他4株分离株中该IVS的位置和大小。这是关于真细菌16S rRNA基因中存在IVS的首次报道。