实时荧光定量PCR直接检测粪便标本艰难梭菌tcdB基因

发布时间：2019-07-31 14:27

【摘要】：目的建立艰难梭菌实时荧光定量PCR检测方法,直接检测粪便标本艰难梭菌tcd B基因,并探讨其应用价值。方法根据艰难梭菌基因组设计tcd B基因特异性引物及探针,用ATCC 43255标准株评价其敏感性,选取艰难梭菌生物学特征相近的临床菌株验证其特异性;收集2015年10-12月临床腹泻患者稀便标本115份,提取基因组总DNA后进行tcd B基因检测,并以产毒培养法为参考方法,探讨该方法的临床诊断价值。结果荧光定量PCR法最低检测限为1×10-3ng,并且仅特异性地扩增产毒艰难梭菌;临床样本评价荧光定量PCR的敏感性、特异性、阳性预测值和阴性预测值分别为75.0%、99.1%、85.7%和98.1%。结论实时荧光定量PCR直接检测稀便标本中的艰难梭菌tcd B基因可用于艰难梭菌相关腹泻的快速诊断。
[Abstract]:Objective to establish a real-time fluorescence quantitative PCR method for the detection of Clostridium perfringens tcd B gene in fecal specimens and to explore its application value. Methods tcd B gene specific primers and probes were designed according to Clostridium vulgaris genome, its sensitivity was evaluated by ATCC 43255 standard strain, and its specificity was verified by clinical strains with similar biological characteristics. 115 stool samples from patients with clinical diarrhea from October to December 2015 were collected and tcd B gene was detected after genomic total DNA was extracted, and the clinical diagnostic value of this method was discussed with the method of toxin production culture as reference. Results the detection limit of fluorescence quantitative PCR was 1 脳 10 鈮，

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