Identification of Helicobacter pylori and the cagA genotype in gastric biopsies using highly sensitive real-time PCR as a new diagnostic tool
Identification of Helicobacter pylori and the cagA genotype in gastric biopsies using highly sensitive real-time PCR as a new diagnostic tool
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DOI：
10.1016/j.femsim.2004.12.011
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发表时间：
2005-06-01
影响因子：
--
通讯作者：
Azuma, T
中科院分区：
文献类型：
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作者：
Yamazaki, S;Kato, S;Azuma, T
CagA蛋白是幽门螺杆菌的毒力因子之一，已观察到CagA的两种主要亚型，即西方型和东亚型。CagA从细菌注射到胃上皮细胞中，经历酪氨酸磷酸化，并与含Src同源2结构域的蛋白酪氨酸磷酸酶SHP-2结合。东亚型CagA与SHP-2的结合比西方型CagA更强。本研究采用高灵敏度的实时荧光定量PCR技术对CagA亚型进行检测，旨在建立一种检测H. pylori和CagA亚型。我们分别在Western特异性序列区和东亚特异性序列区设计了Western或东亚cagA的引物和探针组，并将H. pylori 16S rRNA。我们可以检测到H。pylori 16 S rRNA基因、Western和East Asia-cagA基因。H. pylori感染率为100%。在泰国患者中，87.8%（36/41）为cagA阳性; 26.8%（11/41）为Western-cagA阳性，53.7%（22/41）为东亚-cagA阳性，而7.3%（3/41）对两种类型的cagA均反应。这些结果表明，这种实时荧光PCR系统提供了一个高度敏感的评估CagA型作为一个新的诊断工具，致病性H。幽门感染(c)2005年，欧洲微生物学会联合会。Elsevier B. V.出版，保留所有权利。
The CagA protein is one of the virulence factors of Helicobacter pylori, and two major subtypes of CagA have been observed, the Western and East Asian type. CagA is injected from the bacteria into gastric epithelial cells, undergoes tyrosine phosphorylation, and binds to Src homology 2 domain-containing protein-tyrosine phosphatase SHP-2. The East Asian type CagA binds to SHP-2 more strongly than the Western type CagA. Here, we tried to distinguish the CagA type by highly sensitive real-time PCR with the objective of establishing a system to detect H. pylori and CagA subtypes from gastric biopsies. We designed primers and probe sets for Western or East Asian-cagA at Western-specific or East Asian-specific sequence regions, respectively, and H. pylori 16S rRNA. We could detect the H. pylori 16S rRNA gene, Western and East Asian-cagA gene from DNA of gastric biopsies. The sensitivity and specificity for H. pylori infection was 100% in this system. In Thai patients, 87.8% (36/41) were cagA-positive; 26.8% (11/41) were Western-cagA positive and 53.7% (22/41) were East Asian-cagA positive, while 7.3% (3/41) reacted with both types of cagA. These results suggest that this real-time PCR system provides a highly sensitive assessment of CagA type as a new diagnostic tool for the pathogenicity of H. pylori infection. (c) 2005 Federation of European Microbiological Societies. Published by Elsevier B.V. All rights reserved.