Burnett Tracey A, A Hornitzky Michael, Kuhnert Peter, Djordjevic Steven P
NSW Agriculture, Elizabeth Macarthur Agricultural Institute, PMB 8, Camden, NSW 2570, Australia.
FEMS Microbiol Lett. 2002 Nov 5;216(2):201-9. doi: 10.1111/j.1574-6968.2002.tb11436.x.
Six previously published polymerase chain reaction (PCR) assays each targeting different genes were used to speciate 116 isolates previously identified as Campylobacter jejuni using routine microbiological techniques. Of the 116 isolates, 84 were of poultry origin and 32 of human origin. The six PCR assays confirmed the species identities of 31 of 32 (97%) human isolates and 56 of 84 (67%) poultry isolates as C. jejuni. Twenty eight of 84 (33%) poultry isolates were identified as Campylobacter coli and the remaining human isolate was tentatively identified as Campylobacter upsaliensis based on the degree of similarity of 16S rRNA gene sequences. Four of six published PCR assays showed 100% concordance in their ability to speciate 113 of the 116 (97.4%) isolates; two assays failed to generate a PCR product with four to 10 isolates. A C. coli-specific PCR identified all 28 hippuricase gene (hipO)-negative poultry isolates as C. coli although three isolates confirmed to be C. jejuni by the remaining five assays were also positive in this assay. A PCR-restriction fragment length polymorphism assay based on the 16S rRNA gene was developed, which contrary to the results of the six PCR-based assays, identified 28 of 29 hipO-negative isolates as C. jejuni. DNA sequence analysis of 16S rRNA genes from four hipO-negative poultry isolates showed they were almost identical to the C. jejuni type strain 16S rRNA sequences ATCC43431 and ATCC33560 indicating that assays reliant on 16S rRNA sequence may not be suitable for the differentiation of these two species.
使用六种先前发表的分别针对不同基因的聚合酶链反应（PCR）检测方法，对116株先前采用常规微生物技术鉴定为空肠弯曲菌的分离株进行了菌种鉴定。在这116株分离株中，84株来源于家禽，32株来源于人类。六种PCR检测方法证实，32株人类分离株中的31株（97%）以及84株家禽分离株中的56株（67%）为空肠弯曲菌。84株家禽分离株中的28株（33%）被鉴定为结肠弯曲菌，剩余的一株人类分离株基于16S rRNA基因序列的相似程度被初步鉴定为乌普萨拉弯曲菌。六种已发表的PCR检测方法中的四种在对116株分离株中的113株（97.4%）进行菌种鉴定时显示出100%的一致性；两种检测方法未能对4至10株分离株扩增出PCR产物。一种结肠弯曲菌特异性PCR检测方法将所有28株马尿酸酶基因（hipO）阴性的家禽分离株鉴定为结肠弯曲菌，不过另外五种种检测方法证实为空肠弯曲菌的三株分离株在该检测方法中也呈阳性。开发了一种基于16S rRNA基因的PCR-限制性片段长度多态性检测方法，与六种基于PCR的检测方法的结果相反，该方法将29株hipO阴性分离株中的28株鉴定为空肠弯曲菌。对四株hipO阴性家禽分离株的16S rRNA基因进行DNA序列分析表明，它们与空肠弯曲菌模式菌株16S rRNA序列ATCC43431和ATCC33560几乎相同，这表明依赖16S rRNA序列的检测方法可能不适用于这两个菌种的区分。
