School of Biotechnology, Banaras Hindu University, Varanasi, India.
Department of Gastroenterology, Institute of Medical Sciences, Banaras Hindu University, Varanasi, India.
J Med Microbiol. 2010 Jan;59(Pt 1):32-40. doi: 10.1099/jmm.0.013763-0.
The cag pathogenicity island (cagPAI) has been reported to be the major virulence determinant in Helicobacter pylori-related diseases. In the present study, the diversity of the cagA gene and the integrity of the cagPAI in 158 H. pylori strains from Varanasi (North India) and Hyderabad (South India) were studied by amplifying the cagA gene (approximately 3.5 kb), followed by PCR-RFLP analysis. The results revealed significant differences in the cagA gene and the integrity of the cagPAI between North and South Indian isolates. Of 158 isolates, 40 (34.8 %) from Varanasi and 20 (46.5 %) from Hyderabad were found to carry an intact cagPAI. A partially deleted cagPAI was present in 75 (65.2 %) isolates from Varanasi and 23 (53.5 %) from Hyderabad. None of the isolates showed complete deletion of the cagPAI. Differences in the cagA 5' and 3' regions were also noted, and 11 isolates (8 from Varanasi and 3 from Hyderabad) that were cagA negative with primers for the 5' region turned out to be cagA positive with primers for the 3' variable region. It is tentatively concluded that the 3' variable region may be a better marker for cagA typing. The results also showed that the majority of the isolates harboured the Western-type EPIYA motif. PCR-RFLP analysis of the cagA gene showed 29 distinguishable digestion patterns, and cluster analysis of RFLP types from a random selection of 32 isolates placed all of the isolates into 5 groups. These results demonstrate that significant differences in the cagPAI occur among isolates from North and South India, and that RFLP of cagA could be employed for elucidating genetic variations among various isolates of H. pylori.
细胞毒素相关基因座(cagPAI)已被报道为与幽门螺杆菌相关疾病的主要毒力决定因素。在本研究中,通过扩增 cagA 基因(约 3.5kb),然后进行 PCR-RFLP 分析,研究了来自北印度瓦拉纳西和南印度海得拉巴的 158 株幽门螺杆菌菌株中 cagA 基因的多样性和 cagPAI 的完整性。结果表明,北印度和南印度分离株的 cagA 基因和 cagPAI 的完整性存在显著差异。在 158 株分离株中,来自瓦拉纳西的 40 株(34.8%)和来自海得拉巴的 20 株(46.5%)携带完整的 cagPAI。来自瓦拉纳西的 75 株(65.2%)分离株和来自海得拉巴的 23 株(53.5%)分离株存在部分缺失的 cagPAI。没有分离株显示 cagPAI 的完全缺失。还注意到 cagA 5'和 3'区域的差异,用 5'区域引物检测为 cagA 阴性的 11 株分离株(来自瓦拉纳西的 8 株和来自海得拉巴的 3 株)用 3'可变区引物检测为 cagA 阳性。可以推测,3'可变区可能是 cagA 分型的更好标记。结果还表明,大多数分离株携带西方型 EPIYA 基序。cagA 基因的 PCR-RFLP 分析显示出 29 种可区分的消化模式,对随机选择的 32 株分离株的 RFLP 类型进行聚类分析,将所有分离株分为 5 组。这些结果表明,北印度和南印度分离株的 cagPAI 存在显著差异,cagA 的 RFLP 可用于阐明幽门螺杆菌不同分离株的遗传变异。