一种检测胆螺杆菌套式PCR方法的建立

目的 建立一种敏感性高、特异性强的检测胆螺杆菌的套式PCR方法。方法 基于17种胆螺杆菌亚种的16S rRNA 基因设计筛选出1套套式引物(2条内引物、2条外引物),优化反应条件后,通过模拟粪便标本、动物模型标本和临床标本的检测对方法的敏感性和特异性进行评估。结果 模拟粪便标本中,该方法 检测胆螺杆菌的敏感性达到10 CFU/100 μL。3例感染胆螺杆菌的SPF级BALB/c小鼠模型中,3例小鼠粪便和盲肠中均可检测到胆螺杆菌,1例肝脏标本中检测到了胆螺杆菌。10例胆石病患者中,有2例患者的胆汁、胆囊粘膜和粪便标本中检测到了胆螺杆菌。结论 本研究建立的套式PCR方法 ,敏感性高、特异性强,可用于检测胆螺杆菌的感染。

Development of a nested PCR assay for detection of Helicobacter bilis

In this study, the objective is to establish a nested-PCR assay for the detection of H. bilis with high sensitivity and specificity. The nested primers were designed based on sequences of 16S rRNA gene of seventeen subtypes of H. bilis. After optimizing reaction condition, the sensitivity and specificity of the assay were examined via the detection of feces simulated samples, mice infection model samples and clinic patients’ samples. The detection sensitivity of H. bilis strain for feces simulated samples was 10 CFU/100 μL. H. bilis was successfully detected in the liver, caecum and feces of experimentally infected mice. Moreover, H. bilis was successfully detected in the bile, cholecyst mucous membrane and feces samples from two of ten patients with cholelithiasis. Due to the PCR assay’s high sensitivity and specificity, the method may be used to detect the infection of H. bilis.