Peng H, Pan G, Chao S
Department of Gastroenterology, Peking Union Medical College Hospital, Chinese Academy of Medical Science, Beijing, 100730.
Zhonghua Yi Xue Za Zhi. 1999 Mar;79(3):181-4.
To use PCR/SSCP analysis of the vacuolating cytotoxin gene (vacA) of H. pylori for differentiation of various strains of H. pylori.
PCR was performed using the primers amplifing vacA gene of the bacteria embeded in the gastric mucosa of 159 patients with various gastric duodenal diseases. The products of PCR were further processed for SSCP analysis and southern blot hybridization. In the meantime, vacA genes of three different SSCP-patterns from three patients with duodenal ulcers were sequenced.
The rate of detection of H. pylori with the method was 100%. vacA1 and vacA2, the two subtypes of vacA, were 76.5% (114/149) and 23.5%(35/149), respectively. Eight different SSCP-patterns were distributed in various gastroduodenal diseases, and that 80% of duodenal ulcers was predominated with B pattern. Sequencing of DNA indicated a diversity of vacA gene structure.
PCR/SSCP can be used in the differentiation of different strains of H. pylori in epidemology, and in the follow up study after H. pylori eradication, especially in the differentiation between H. pylori recrudescence and reinfection.
运用聚合酶链反应/单链构象多态性分析(PCR/SSCP)检测幽门螺杆菌空泡毒素基因(vacA),以区分不同菌株的幽门螺杆菌。
采用引物对159例患有各种胃十二指肠疾病患者胃黏膜中幽门螺杆菌的vacA基因进行PCR扩增。PCR产物进一步进行SSCP分析和Southern印迹杂交。同时,对3例十二指肠溃疡患者的3种不同SSCP模式的vacA基因进行测序。
该方法检测幽门螺杆菌的阳性率为100%。vacA的两个亚型vacA1和vacA2分别占76.5%(114/149)和23.5%(35/149)。8种不同的SSCP模式分布于各种胃十二指肠疾病中,十二指肠溃疡中80%以B模式为主。DNA测序表明vacA基因结构具有多样性。
PCR/SSCP可用于幽门螺杆菌不同菌株的流行病学鉴别,以及幽门螺杆菌根除后的随访研究,尤其是用于区分幽门螺杆菌复发和再感染。