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首页> 外文期刊>Journal of Clinical Microbiology >Real-Time LightCycler PCR for Detection and Discrimination of Bordetella pertussis and Bordetella parapertussis
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Real-Time LightCycler PCR for Detection and Discrimination of Bordetella pertussis and Bordetella parapertussis

机译：实时LightCycler PCR检测和鉴别百日咳博德特氏菌和副百日咳博德特氏菌

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摘要

Real-time PCR assays based on the LightCycler technology were developed for individual (simplex PCR) and simultaneous (duplex PCR) detection and discrimination of Bordetella pertussis and Bordetella parapertussis in clinical samples. The assays were evaluated with 113 specimens from patients with and without symptoms of pertussis. Results were compared to those from conventional culture and TaqMan real-time PCR. The analytical sensitivity ranged from 0.1 to 10 CFU for B. pertussis and B. parapertussis, and intra- and interassay variations were less than 7%. Results were available within 2 h. With the simplex format, 21 of 100 samples from patients with clinical symptoms of pertussis were positive for B. pertussis and/or B. parapertussis. With the duplex format, 18 of 100 samples were positive. LightCycler PCR increased the diagnostic sensitivity over that of culture by 2.0-fold (duplex PCR) (P = 0.08) to 2.3-fold (simplex PCR) (P = 0.02). Our data suggest that duplex PCR in this format showed good analytical sensitivity but lost some sensitivity on clinical samples compared with the simplex format.
机译：开发了基于LightCycler技术的实时PCR检测试剂盒，用于临床样品中百日咳博德特氏菌和副百日咳博德特氏菌的分别（简单PCR）和同时（双重PCR）检测和鉴别。 。使用来自和不带有百日咳症状的患者的113个标本对测定进行了评估。将结果与常规培养和TaqMan实时PCR的结果进行比较。对 B 的分析灵敏度范围为0.1到10 CFU。 百日咳和 B 。 百日咳，测定内和测定间差异小于7％。 2小时内可获得结果。采用单纯形格式，百日咳临床症状患者的100个样本中有21个 B 阳性。 百日咳和/或 B 。 百日咳。使用双工格式时，100个样本中有18个呈阳性。 LightCycler PCR使培养物的诊断灵敏度比培养物高2.0倍（双重PCR）（ P = 0.08）至2.3倍（单纯PCR）（ P = 0.02） 。我们的数据表明，与单纯形相比，这种形式的双链PCR显示出良好的分析灵敏度，但对临床样品失去了某些敏感性。

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