【摘要】：目的建立艱難梭菌實(shí)時(shí)熒光定量PCR檢測(cè)方法,直接檢測(cè)糞便標(biāo)本艱難梭菌tcd B基因,并探討其應(yīng)用價(jià)值。方法根據(jù)艱難梭菌基因組設(shè)計(jì)tcd B基因特異性引物及探針,用ATCC 43255標(biāo)準(zhǔn)株評(píng)價(jià)其敏感性,選取艱難梭菌生物學(xué)特征相近的臨床菌株驗(yàn)證其特異性;收集2015年10-12月臨床腹瀉患者稀便標(biāo)本115份,提取基因組總DNA后進(jìn)行tcd B基因檢測(cè),并以產(chǎn)毒培養(yǎng)法為參考方法,探討該方法的臨床診斷價(jià)值。結(jié)果熒光定量PCR法最低檢測(cè)限為1×10-3ng,并且僅特異性地?cái)U(kuò)增產(chǎn)毒艱難梭菌;臨床樣本評(píng)價(jià)熒光定量PCR的敏感性、特異性、陽(yáng)性預(yù)測(cè)值和陰性預(yù)測(cè)值分別為75.0%、99.1%、85.7%和98.1%。結(jié)論實(shí)時(shí)熒光定量PCR直接檢測(cè)稀便標(biāo)本中的艱難梭菌tcd B基因可用于艱難梭菌相關(guān)腹瀉的快速診斷。
[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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