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首页> 外文期刊>Journal of Clinical Microbiology >Perirectal Swab Surveillance for Clostridium difficile by Use of Selective Broth Preamplification and Real-Time PCR Detection of tcdB
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Perirectal Swab Surveillance for Clostridium difficile by Use of Selective Broth Preamplification and Real-Time PCR Detection of tcdB

机译:选择性培养液预扩增和tcdB实时PCR检测对艰难梭菌的直肠拭子监测

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摘要

Active surveillance testing to identify and isolate asymptomatic carriers of toxigenic Clostridium difficile has been limited by the lack of a test that is sensitive, specific, and timely enough to serve as an infection control tool. We tested DNA preamplified from perirectal surveillance specimens in a liquid medium selective for C. difficile by using a modified commercial real-time PCR assay. All fermenting specimens were subcultured, and isolates were tested for toxigenicity. Culture-positive toxigenic isolates served as the gold standard for comparison with the broth preamplification/PCR assay. The limit of detection for the assay was 1 CFU. Relative to toxigenic anaerobic culture, the sensitivity, specificity, and positive and negative predictive values of this assay were 70/70 (100.0%), 422/426 (99.1%), 70/74 (94.6%), and 422/422 (100.0%), respectively. These data demonstrate that selective broth preamplification and real-time PCR of perirectal swab specimens constitute a practical approach to the detection of asymptomatic C. difficile carriage.
机译:由于缺乏足够灵敏,特异,及时的感染检测工具,主动识别和鉴别无毒的梭状芽胞杆菌无症状携带者的主动监视测试受到了限制。我们通过使用改良的商业实时PCR分析法,在对艰难梭菌有选择性的液体培养基中测试了从直肠周围监视标本中预扩增的DNA。所有发酵标本都进行了传代培养,并对分离物的产毒性进行了测试。培养阳性产毒分离物用作与肉汤预扩增/ PCR测定法进行比较的金标准。该测定的检测极限是1 CFU。相对于有毒厌氧培养,此测定法的灵敏度,特异性以及阳性和阴性预测值分别为70/70(100.0%),422/426(99.1%),70/74(94.6%)和422/422( 100.0%)。这些数据表明,直肠周围拭子标本的选择性肉汤预扩增和实时PCR构成了检测无症状艰难梭菌携带的实用方法。

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