Tatti Kathleen M, Wu Kai-Hui, Tondella Maria Lucia, Cassiday Pamela K, Cortese Margaret M, Wilkins Patricia P, Sanden Gary N
Meningitis and Vaccine Preventable Diseases Branch, Division of Bacterial Diseases, National Center for Immunization and Respiratory Diseases, Centers for Disease Control and Prevention, Atlanta, GA 30333, USA.
Diagn Microbiol Infect Dis. 2008 Jul;61(3):264-72. doi: 10.1016/j.diagmicrobio.2008.02.017. Epub 2008 Apr 25.
Novel, highly specific, and sensitive real-time polymerase chain reaction (PCR) assays using 2 targets, insertion sequence (IS481) and pertussis toxin subunit 1 (ptxS1), were developed to detect Bordetella pertussis and to differentiate between relevant Bordetella spp. Sixty-four non-Bordetella isolates were negative by both assays, demonstrating the specificity of the assays. B. pertussis, Bordetella parapertussis, and Bordetella holmesii isolates were specifically identified using the assays. The lower limit of detection was less than 10 genomic equivalents per reaction for the IS481 and ptxS1 assays. These assays were evaluated using 145 human clinical specimens obtained during cough-illness outbreak investigations, and PCR results were compared with Bordetella spp. culture results. Twenty-seven (18.6%) specimens had late positive cycle threshold (Ct) values (35 <or= Ct < 40) using the IS481 assay with corresponding negative results using the ptxS1 assay and culture and were considered indeterminate. Guidelines for use of PCR testing and interpretation of results during cough-illness outbreaks are discussed.
开发了使用插入序列（IS481）和百日咳毒素亚基1（ptxS1）这两个靶标的新型、高特异性和高灵敏度实时聚合酶链反应（PCR）检测方法，用于检测百日咳博德特氏菌并区分相关博德特氏菌属。64株非博德特氏菌分离株在两种检测方法中均为阴性，证明了检测方法的特异性。使用这些检测方法可特异性鉴定百日咳博德特氏菌、副百日咳博德特氏菌和霍氏博德特氏菌分离株。IS481和ptxS1检测方法的检测下限均低于每个反应10个基因组当量。使用在咳嗽疾病暴发调查期间获得的145份人类临床标本对这些检测方法进行了评估，并将PCR结果与博德特氏菌属培养结果进行了比较。使用IS481检测方法时，27份（18.6%）标本的循环阈值（Ct）值较晚（35≤Ct＜40），而使用ptxS1检测方法和培养时结果为阴性，这些标本被视为不确定。文中讨论了咳嗽疾病暴发期间PCR检测的使用指南和结果解读。
