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基于尿素酶C基因PCR的幽门螺杆菌感染诊断及胃活检组织中细胞毒素相关基因A的检测

UreC PCR based diagnosis of Helicobacter pylori infection and detection of cag A gene in gastric biopsies.

作者信息

Mishra K K, Srivastava S, Dwivedi P P, Prasad K N, Ayyagari A

机构信息

Department of Microbiology, Sanjay Gandhi Postgraduate Institute of Medical Science, Lucknow, India.

出版信息

Indian J Pathol Microbiol. 2002 Jan;45(1):31-7.

Abstract

Polymerase chain reaction assay using ureC gene specific primers for the detection of Helicobacter pylori in gastric biopsy specimens from 116 dyspeptic patients was compared with other routine invasive diagnostic methods (culture, rapid urease test [RUT] and histology). In parallel, gastric biospy specimens from 54 patients and their corresponding Helicobacter pylori isolates were subjected to PCR with cagA targeting primers using standard protocols. Helicobacter pylori were detected in 53%, 43%, 48% and 50% of patients by PCR, RUT, culture and histological examination respectively. Based on histology and culture positive and at least three test positive result, 44 (37%), 46 (39%) and 26 (22%), and 56 (48%), 52 (44%) and 8 (6%) patients were classified as Helicobacter pylori positive, negative and indeterminate respectively. The sensitivity and specificity of PCR assay was the highest-95% and 100% when compared with both culture and histology positive, and at least any three positive results respectively. The result of cagA positivity in 54 gastric biopsy specimens and their corresponding Helicobacter pylori isolates were identical; 18 of 20 (90%) duodenal ulcer patients and 23 of 28 (82%) patients with chronic gastritis and 2 (40%) of 5 patients with portal hypertension and one gastric biopsy specimens from gastric cancer patients were found to be cagA positive. PCR-based method to detect Helicobacter pylori and the virulence gene cag A directly from gastric biopsy specimens appears to be promising and can curtail the lengthy process of culture-based approaches. The procedure proved to be rapid and reliable and could be utilized for diagnostic purposes.

摘要

使用ureC基因特异性引物的聚合酶链反应检测法,对116例消化不良患者的胃活检标本进行幽门螺杆菌检测,并与其他常规侵入性诊断方法(培养、快速尿素酶试验[RUT]和组织学检查)进行比较。同时,采用标准方案,对54例患者的胃活检标本及其相应的幽门螺杆菌分离株进行cagA靶向引物的PCR检测。通过PCR、RUT、培养和组织学检查分别在53%、43%、48%和50%的患者中检测到幽门螺杆菌。根据组织学和培养阳性以及至少三项检测阳性结果,分别将44例(37%)、46例(39%)和26例(22%)以及56例(48%)、52例(44%)和8例(6%)患者分类为幽门螺杆菌阳性、阴性和不确定。与培养和组织学阳性以及至少任何三项阳性结果相比,PCR检测法的敏感性和特异性最高,分别为95%和100%。54例胃活检标本及其相应的幽门螺杆菌分离株的cagA阳性结果相同;20例十二指肠溃疡患者中有18例(90%)、28例慢性胃炎患者中有23例(82%)、5例门静脉高压患者中有2例(40%)以及1例胃癌患者的胃活检标本被发现cagA阳性。基于PCR的方法直接从胃活检标本中检测幽门螺杆菌和毒力基因cagA似乎很有前景,并且可以缩短基于培养方法的漫长过程。该方法被证明快速可靠,可用于诊断目的。

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