Müller F M, Schnitzler N, Cloot O, Kockelkorn P, Haase G, Li Z
Children's Hospital, Institute for Medical Microbiology, University of Aachen, Germany.
Diagn Microbiol Infect Dis. 1998 Aug;31(4):517-23. doi: 10.1016/s0732-8893(98)00043-1.
The inclusion of an appropriate internal control DNA in polymerase chain reaction (PCR) is a rapid and simple method for the detection of PCR failure. Two PCR coamplification internal control DNAs (ICD I and ICD II) with the same primer-binding sequences as the target DNA for the detection of Bordetella pertussis and Bordetella parapertussis were produced using an overlap extension technique and a PCR MIMIC construction kit, respectively. The ICD II was further evaluated in a prospective clinical study in 360 patients with a clinical diagnosis of pertussis. From 360 nasopharyngeal swabs the internal control was positive in 318 (88%) samples, but was negative in 42 (12%). After phenol-chloroform extraction an additional 10 internal controls became positive. For the detection of PCR failure, the use of internal control DNA is highly recommended for PCR-based identification of B. pertussis and B. parapertussis organisms from nasopharyngeal swabs and aspirates.
在聚合酶链反应（PCR）中加入合适的内部控制DNA是检测PCR失败的一种快速简便的方法。分别使用重叠延伸技术和PCR MIMIC构建试剂盒制备了两种与百日咳博德特氏菌和副百日咳博德特氏菌检测的靶DNA具有相同引物结合序列的PCR共扩增内部控制DNA（ICD I和ICD II）。在一项对360例临床诊断为百日咳的患者进行的前瞻性临床研究中，对ICD II进行了进一步评估。在360份鼻咽拭子中，内部控制在318份（88%）样本中呈阳性，但在42份（12%）样本中呈阴性。经过酚-氯仿提取后，又有10个内部控制呈阳性。为了检测PCR失败，强烈建议在基于PCR从鼻咽拭子和吸出物中鉴定百日咳博德特氏菌和副百日咳博德特氏菌时使用内部控制DNA。
