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聚合酶链反应检测粪便和唾液中幽门螺杆菌DNA:综述

Detection of Helicobacter pylori DNA in feces and saliva by polymerase chain reaction: a review.

作者信息

Kabir Shahjahan

机构信息

Academic Research and Information Management, Stockholm, Sweden.

出版信息

Helicobacter. 2004 Apr;9(2):115-23. doi: 10.1111/j.1083-4389.2004.00207.x.

Abstract

The polymerase chain reaction (PCR), known for its high sensitivity and specificity, has been used for the detection of Helicobacter pylori DNA in bodily materials such as feces and saliva. Since fecal specimens contain PCR inhibitors, DNA before PCR amplification has been purified using various biochemical, immunological and physical pre-PCR steps. Several PCR protocols, differing from each other in the selection of genomic targets and primers, have produced varying degrees of specificity and sensitivity in detecting H. pylori DNA. PCR identified antimicrobial resistance of H. pylori in feces. It also detected virulence factor genes such as the cytotoxin-associated gene (cagA) and vacuolating cytotoxin gene (vacA) in feces and saliva. While the cagA gene was detected in 50-60% of fecal specimens, it was found in 25% of salivary specimens from patients. There was considerable variation in the detection rate of H. pylori DNA in salivary samples. The detection rate in saliva with the most effective primer pair was lower than that observed in feces, making saliva a less suitable specimen for the diagnosis of H. pylori infection. There is controversy regarding the permanent presence of H. pylori in saliva. Whether the salivary and gastric specimens of an individual harbor identical or different strains has not been resolved. PCR cannot distinguish between living and dead organisms. However, it can offer quick results on fecal and salivary specimens, which may contain fastidious and slow-growing H. pylori in low numbers.

摘要

聚合酶链反应(PCR)以其高灵敏度和特异性而闻名,已被用于检测粪便和唾液等身体物质中的幽门螺杆菌DNA。由于粪便标本含有PCR抑制剂,在PCR扩增之前,已使用各种生化、免疫和物理的PCR前处理步骤对DNA进行了纯化。几种PCR方案在基因组靶点和引物的选择上彼此不同,在检测幽门螺杆菌DNA时产生了不同程度的特异性和灵敏度。PCR鉴定了粪便中幽门螺杆菌的抗菌耐药性。它还在粪便和唾液中检测到了毒力因子基因,如细胞毒素相关基因(cagA)和空泡毒素基因(vacA)。虽然在50%-60%的粪便标本中检测到了cagA基因,但在患者的唾液标本中仅25%检测到该基因。唾液样本中幽门螺杆菌DNA的检测率存在相当大的差异。使用最有效的引物对时,唾液中的检测率低于粪便中的检测率,这使得唾液不太适合作为诊断幽门螺杆菌感染的标本。关于幽门螺杆菌是否长期存在于唾液中存在争议。个体的唾液和胃标本中携带的菌株是否相同或不同尚未得到解决。PCR无法区分活的和死的生物体。然而,它可以快速给出粪便和唾液标本的检测结果,这些标本中可能含有数量较少、生长缓慢且挑剔的幽门螺杆菌。

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