Qiao Wen, Hu Jia-Lu, Xiao Bing, Wu Kai-Chun, Peng Dao-Rong, Atherton John C, Xue Hui
Department of Gastroenterology, First Hospital, Xi'an Jiaotong University, Xi'an 710061, Shaanxi Province, China. xhy1202@ sohu.com
World J Gastroenterol. 2003 Aug;9(8):1762-6. doi: 10.3748/wjg.v9.i8.1762.
To establish stock of clinical Helicobacter pylori (H. pylori) isolates, to perform cagA and vacA typing of these isolates, to evaluate the relationship between genotypes of cagA and vacA and upper gastrointestinal diseases and to assess the association of vacA genotypes with presence of the pathogenicity marker-cagA.
Clinical H.pylori strains were isolated from the antrum of 259 patients in Clumbia agar. The isolated H.pylori strains were identified by histology, and16SrRNA PCR. CagA genotypes were detected by colony hybridization, the probe was derived from the cloned plasmid PcagA, and digested by EcoRI-HindIII and the isolated PcagA DNA fragment was radioactively labelled by the random priming method. vacA genes types (s,m)and subtypes (s1a, s1b, s2) were typed by PCR. Vacuolating toxin was detected with neutral red absorb test. The results were treated statistically by chi(2) test, t test, and rank sum test.
A total of 192 clinical H.pylori strains were isolated and the stock of Helicobacter pylori was established. The total positive rate of cagA was 87 % in all gastric diseases, and 95 % in gastric cancer group. There was a difference between gastric cancer group and the other groups (P<0.05) except duodenal ulcer group. The expression of type s1 of vacA was more than type s2 (P<0.05), and, the expression of type m1 was equal to type m2. In gastric cancer group, there was a difference between s1a and s1b (P<0.05), and s1a was more than s1b. Vacuolating toxins were more in Xi'an area isolates.
The cagA(+) vacA type s1 clinical isolates are more in Xi'an area, but this can not serve as an index to predict gastric cancer.
建立临床幽门螺杆菌(H. pylori)分离株库,对这些分离株进行cagA和vacA分型,评估cagA和vacA基因型与上消化道疾病之间的关系,并评估vacA基因型与致病性标志物cagA存在之间的关联。
从259例患者的胃窦部在哥伦比亚琼脂培养基上分离临床H.pylori菌株。通过组织学和16SrRNA PCR鉴定分离出的H.pylori菌株。通过菌落杂交检测CagA基因型,探针源自克隆质粒PcagA,用EcoRI-HindIII消化,分离的PcagA DNA片段通过随机引物法进行放射性标记。通过PCR对vacA基因类型(s,m)和亚型(s1a,s1b,s2)进行分型。用中性红吸收试验检测空泡毒素。结果采用卡方检验、t检验和秩和检验进行统计学处理。
共分离出192株临床H.pylori菌株,建立了幽门螺杆菌库。所有胃病中cagA的总阳性率为87%,胃癌组为95%。除十二指肠溃疡组外,胃癌组与其他组之间存在差异(P<0.05)。vacA s1型的表达多于s2型(P<0.05),m1型和m2型的表达相等。在胃癌组中,s1a和s1b之间存在差异(P<0.05),且s1a多于s1b。西安地区分离株中的空泡毒素更多。
西安地区cagA(+) vacA s1型临床分离株较多,但这不能作为预测胃癌的指标。