Giesendorf B A, Quint W G
Academic Hospital Leiden, Department of Medical Microbiology, The Netherlands.
Cell Mol Biol (Noisy-le-grand). 1995 Jul;41(5):625-38.
Since Campylobacters have fastidious growth requirements and conventional detection and identification requires at least 4-6 days, the development of fast but reliable detection procedures is needed. Although methods based on DNA probe technology have been developed, these are not sensitive enough for the detection of Campylobacter spp. in food products. Therefore a PCR procedure based on the amplification of the 16S rRNA gene was developed that specifically detects the thermophilic Campylobacter species. This assay provides an excellent tool for the rapid and sensitive isolation and identification of Campylobacter spp. from chicken samples. In order to further identify the different Campylobacter spp., which are difficult to distinguish by conventional methods, PCR mediated approximately DNA typing was used to select species-specific DNA probes. This combination of PCR fingerprinting and probe hybridization results in a highly specific identification assay and provides an example of specific test development without the prior need for DNA sequence information. PCR mediated DNA typing was also used to study the epidemiology of diarrheal diseases caused by Campylobacter spp. Using primers complementary to dispersed repetitive DNA sequences and arbitrarily chosen DNA motifs PCR fingerprinting has proven to be a fast, highly discriminative and relatively simple method that can be applied in epidemiological investigations on Campylobacter infections. Besides this application of PCR fingerprinting for typing of Campylobacter spp. this method can also be used for the development of specific DNA probes.
由于弯曲杆菌生长要求苛刻，传统的检测和鉴定至少需要4 - 6天，因此需要开发快速且可靠的检测方法。尽管基于DNA探针技术的方法已经开发出来，但这些方法对于食品中弯曲杆菌属的检测不够灵敏。因此，开发了一种基于16S rRNA基因扩增的PCR程序，该程序可特异性检测嗜热弯曲杆菌种。该检测方法为从鸡肉样品中快速、灵敏地分离和鉴定弯曲杆菌属提供了一个极好的工具。为了进一步鉴定传统方法难以区分的不同弯曲杆菌属，使用PCR介导的近似DNA分型来选择种特异性DNA探针。PCR指纹图谱和探针杂交的这种结合产生了一种高度特异性的鉴定检测方法，并提供了一个无需事先获得DNA序列信息即可开发特异性检测方法的实例。PCR介导的DNA分型还用于研究由弯曲杆菌属引起的腹泻病的流行病学。使用与分散的重复DNA序列互补的引物和任意选择的DNA基序，PCR指纹图谱已被证明是一种快速、高度鉴别且相对简单的方法，可应用于弯曲杆菌感染的流行病学调查。除了将PCR指纹图谱用于弯曲杆菌属的分型外，该方法还可用于开发特异性DNA探针。
