Nygren M, Reizenstein E, Ronaghi M, Lundeberg J
Department of Biotechnology, KTH, Royal Institute of Technology, Sweden.
J Clin Microbiol. 2000 Jan;38(1):55-60. doi: 10.1128/JCM.38.1.55-60.2000.
The pertussis toxin (PT) promoter region is a frequently used target for DNA-based diagnosis of pertussis and parapertussis infections. The reported polymorphism in this region has also allowed discrimination of species in mixtures with several Bordetella species by their specific PCR amplicon restriction patterns. In the present study, we investigated the degree of polymorphism in order to confirm the reliability of the assay. Five different sequence types of the amplified 239- or 249-bp region were found among the 33 Bordetella pertussis, B. parapertussis, and B. bronchiseptica American Type Culture Collection reference strains and patient isolates analyzed. According to the sequences that were obtained and according to the PT promoter sequences already available in the databases, restriction enzyme analysis with TaqI, BglI, and HaeII, which gave four different patterns, can be performed to reliably identify B. pertussis, B. parapertussis, and B. bronchiseptica.
百日咳毒素(PT)启动子区域是基于DNA诊断百日咳和副百日咳感染常用的靶标。该区域报道的多态性也使得通过特定的PCR扩增子限制性图谱,能够区分与几种博德特氏菌属物种混合样本中的物种。在本研究中,我们调查了多态性程度,以确认该检测方法的可靠性。在所分析的33株百日咳博德特氏菌、副百日咳博德特氏菌和支气管败血博德特氏菌美国模式培养物保藏中心参考菌株及患者分离株中,发现扩增的239或249 bp区域有5种不同的序列类型。根据获得的序列以及数据库中已有的PT启动子序列,可以用TaqI、BglI和HaeII进行限制性酶切分析(产生四种不同模式),以可靠地鉴定百日咳博德特氏菌、副百日咳博德特氏菌和支气管败血博德特氏菌。