Unreliability of Results of PCR Detection of Helicobacter pylori in Clinical or Environmental Samples
Unreliability of Results of PCR Detection of Helicobacter pylori in Clinical or Environmental Samples
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DOI：
10.1128/jcm.01563-08
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发表时间：
2009-03-01
影响因子：
9.4
通讯作者：
Graham, David Y.
中科院分区：
文献类型：
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作者：
Sugimoto, Mitsushige;Wu, Jeng-Yih;Graham, David Y.
这项研究的目的是比较已发表的幽门螺杆菌引物对在胃活检标本和唾液样本中可靠检测幽门螺杆菌的能力。确定了用于检测临床标本中幽门螺杆菌DNA的26对聚合酶链式反应引物的检出限。使用50例幽门螺杆菌阳性和阴性(通过培养和组织学一致)编码的胃活检标本，使用检测下限为100cfu/ml的引物来确定敏感性和特异性。这些结果随后被胃活检标本和确诊为幽门螺杆菌状态的患者的唾液证实。其中5对(HP64-f/HP64-r、Hp1/HP2、EHc-U/EHc-L、VAG-F/VAG-R和ICT37/ICT38)在胃组织中的检出限为100cfu/ml。没有一个有100%的特异性或敏感性；所有的结果都是假阳性。尿素的HP64-f/HP64-r和16S rRNA的HP1/HP2对胃活检标本的敏感性和特异性分别为90%和90%。没有任何组合的引物组合改善了结果。使用这5对引物，54%的阳性唾液样本被确定为假阳性；HP64-f/HP64-r和HP1/HP2两个组合对唾液都产生了假阳性。我们的结论是，临床医生不应该仅仅依靠目前的聚合酶链式反应引物的结果来确定单个患者的幽门螺杆菌状态，或者作为治疗决定的基础。基于环境样本中幽门螺杆菌的聚合酶链式反应鉴定的研究结果应谨慎看待。可能的是，基于多个假定的毒力因子基因的存在，可以识别特定的引物组。
The aim of this study was to compare published Helicobacter pylori primer pairs for their ability to reliably detect H. pylori in gastric biopsy specimens and salivary samples. Detection limits of the 26 PCR primer pairs previously described for detection of H. pylori DNA in clinical samples were determined. Sensitivity and specificity were determined using primers with detection limits of < 100 CFU/ml using 50 H. pylori-positive and -negative (by concordance by culture and histology) coded gastric biopsy specimens. These results were then confirmed with gastric biopsy specimens and saliva from patients with confirmed H. pylori status. Five of the twenty-six previously reported primer pairs (HP64-f/HP64-r, HP1/HP2, EHC-U/EHC-L, VAG-F/VAG-R, and ICT37/ICT38) had detection limits of < 100 CFU/ml in the presence of gastric tissue. None had 100% specificity or sensitivity; all produced false-positive results. The HP64-f/HP64-r for ureA and HP1/HP2 for 16S rRNA individually had sensitivities and specificities of > 90% with gastric biopsy specimens. No combinations of primer pairs improved the results. Using these five primer pairs, 54% of the positive saliva samples were determined to be false positive; both the HP64-f/HP64-r and the HP1/HP2 sets produced false positives with saliva. We conclude that clinicians should not rely on results using current PCR primers alone to decide the H. pylori status of an individual patient or as a basis for treatment decisions. The results of studies based on PCR identification of H. pylori in environmental samples should be viewed with caution. Possibly, specific primers sets can be identified based on the presence of multiple putative virulence factor genes.