Zhang Mao-jun, Gu Yi-xin, Ran Lu, Zhang Jian-zhong
National Institute for Communicable Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 102206, China.
Zhonghua Liu Xing Bing Xue Za Zhi. 2007 Apr;28(4):377-80.
This study was to simultaneously identify Campylobacter jejuni and Campylobacter coli isolates in China by Multi-PCR assay and to study the prevalence of six virulence and toxin genes on them.
A multi-PCR method with three sets of primers specifically designed for application of a 16S rRNA as a universal control, mapA, ceuE based on the specific sequence of C. jejuni and C. coli, was applied to detect 65 Campylobacter isolates from China. Another two separately PCR Primers were directed towards the hippuricase gene (hipO) characteristic of C.jejuni and glyA gene characteristic of C. coli were performed for further confirmation. The presence of the cadF, virB11, flaA, cdtA, cdtB, cdtC genes among these 65 strains were investigated by PCR.
From multi-PCR detection, 42 isolates belonged to C. jejuni, other 23 isolates belong to C. coli. Data showing the identification were 100% in concordance with the separated PCR for hipO and glyA amplification. The efficiency (100%) of identification by these three primers multi-PCR method was higher than the biochemical test (83.1%). The cadF and flaA genes were detected from 100% (65/65) of the isolates and the PCR product of each gene were identical with each isolate. Only 10.8% (7/65) of the isolates were positive for virB11. The cdtA gene was found in 92% (60/65) of the isolates. 97.6% (41/42) of C. jejuni had cdtB gene, whereas no PCR product with this primers for all the C. coli isolates. cdtC was presented in all the isolates but the lengths of PCR products were different. For C. jejuni, it was 555 bp, for C. coli, it was about 465 bp.
This three primers simultaneous multi-PCR method seemed to be useful for the identification of C. jejuni and C. coli isolates from China since cadF and flaA genes were widely spread in Campylobacter isolates in this country. The present report on virB11 was similar to previous reports from other countries, but the distribution of cdt gene cluster in Campylobacter species isolated from China might be different.
本研究旨在通过多重聚合酶链反应（Multi-PCR）法同时鉴定中国的空肠弯曲菌和结肠弯曲菌分离株，并研究其上六种毒力和毒素基因的流行情况。
应用一种多重聚合酶链反应方法，该方法有三组引物，分别基于空肠弯曲菌和结肠弯曲菌的特定序列专门设计用于以16S rRNA作为通用对照、mapA、ceuE，用于检测来自中国的65株弯曲菌分离株。另外针对空肠弯曲菌特有的马尿酸酶基因（hipO）和结肠弯曲菌特有的甘氨酸A基因（glyA）分别设计两组PCR引物进行进一步确认。通过PCR研究这65株菌株中cadF、virB11、flaA、cdtA、cdtB、cdtC基因的存在情况。
通过多重聚合酶链反应检测，42株分离株属于空肠弯曲菌，另外23株分离株属于结肠弯曲菌。显示鉴定结果的数据与用于hipO和glyA扩增的单独PCR结果100%一致。这三种引物多重聚合酶链反应方法的鉴定效率（100%）高于生化试验（83.1%）。100%（65/65）的分离株检测到cadF和flaA基因，每个基因的PCR产物与每个分离株相同。仅10.8%（7/65）的分离株virB11呈阳性。92%（60/65）的分离株中发现了cdtA基因。97.6%（41/42）的空肠弯曲菌有cdtB基因，而所有结肠弯曲菌分离株用该引物均未得到PCR产物。所有分离株中均存在cdtC，但PCR产物长度不同。对于空肠弯曲菌，其长度为555 bp，对于结肠弯曲菌，其长度约为465 bp。
这种三种引物同时进行的多重聚合酶链反应方法似乎对鉴定来自中国的空肠弯曲菌和结肠弯曲菌分离株有用，因为cadF和flaA基因在该国的弯曲菌分离株中广泛传播。关于virB11的本报告与其他国家以前的报告相似，但从中国分离的弯曲菌物种中cdt基因簇的分布可能不同。
