Nakao H, Popovic T
Division of Bacterial and Mycotic Diseases, National Center for Infectious Disease, Centers for Disease Control and Prevention, Atlanta, Georgia 30333, USA.
J Clin Microbiol. 1997 Jul;35(7):1651-5. doi: 10.1128/jcm.35.7.1651-1655.1997.
PCR has proved to be a reliable tool for the detection of the diphtheria toxin gene, tox, and its use has allowed for the rapid differentiation between toxigenic and nontoxigenic strains. In this study, this PCR was further developed, evaluated, and standardized to detect this gene directly from clinical specimens. Optimal conditions for collection, transport, and storage of the clinical specimens and isolation and purification of DNA from the clinical specimens were defined. With two sets of primers that detect the A and B subunits of the diphtheria toxin gene, sensitivity levels of 50 and 500 CFU/PCR mixture, respectively, were achieved. This PCR was evaluated with 162 clinical samples collected from patients with diphtheria and other upper respiratory tract infections, as well as from healthy individuals.
聚合酶链反应(PCR)已被证明是检测白喉毒素基因tox的可靠工具,其应用使得产毒菌株和非产毒菌株得以快速区分。在本研究中,对该PCR方法进行了进一步改进、评估和标准化,以便直接从临床标本中检测该基因。确定了临床标本采集、运输和储存以及从临床标本中分离和纯化DNA的最佳条件。使用两组检测白喉毒素基因A和B亚基的引物,分别达到了50和500 CFU/PCR混合物的灵敏度水平。用从白喉患者和其他上呼吸道感染患者以及健康个体采集的162份临床样本对该PCR进行了评估。