Watanabe Pinhata Juliana Maira, Cergole-Novella Maria Cecilia, Moreira Dos Santos Carmo Andreia, Ruivo Ferro E Silva Regina, Ferrazoli Lucilaine, Tavares Sacchi Claudio, Siqueira de Oliveira Rosangela
Department of Tuberculosis and Mycobacteriosis, Center of Bacteriology, Instituto Adolfo Lutz, Av. Dr Arnaldo, 355, 01246-902 São Paulo, SP, Brazil.
Center of Regional Laboratory of Santo Andre, Rua Ramiro Colleone, 240, 09040-160 Santo André, SP, Brazil.
J Med Microbiol. 2015 Sep;64(9):1040-1045. doi: 10.1099/jmm.0.000121. Epub 2015 Jul 9.
Tuberculosis (TB) is an infectious disease of global distribution, constituting a serious public health problem in Brazil. São Paulo State, located in the south-east of Brazil, notified 16,580 new TB cases in 2013. The Instituto Adolfo Lutz is a public health reference laboratory for TB diagnosis for all the State. Considering that rapid and accurate diagnosis is essential for TB control, the aim of this study was to evaluate the use of an in-house real-time (RT)-PCR assay targeting the mpt64 gene in the routine diagnosis of TB, and to compare this technique with smear microscopy and culture. From August 2012 to October 2013, 715 sputum samples from 657 patients were included in the study. Smear microscopy, culture, phenotypic and PRA-hsp65 identification of mycobacteria, and mpt64 RT-PCR were performed. With respect to confirmed TB cases (n = 62/657; 9.4%), smear microscopy had a sensitivity of 82.3%. Culture and RT-PCR showed the same sensitivity, i.e. 90.3%. Specificity was 99.7, 99.4 and 98.6% for smear microscopy, culture and RT-PCR, respectively. mpt64 RT-PCR showed high sensitivity and specificity for the detection of Mycobacterium tuberculosis complex in sputum samples. This technique can be deployed in laboratories that do not have a rapid test for TB available, enabling the performance of TB diagnosis in up to 5 h.
结核病（TB）是一种全球流行的传染病，在巴西构成严重的公共卫生问题。圣保罗州位于巴西东南部，2013年报告了16580例新的结核病病例。阿道夫·卢茨研究所是该州结核病诊断的公共卫生参考实验室。鉴于快速准确的诊断对结核病控制至关重要，本研究旨在评估针对mpt64基因的内部实时（RT）-PCR检测法在结核病常规诊断中的应用，并将该技术与涂片显微镜检查和培养法进行比较。2012年8月至2013年10月，本研究纳入了657例患者的715份痰标本。进行了涂片显微镜检查、培养、分枝杆菌的表型和PRA-hsp65鉴定以及mpt64 RT-PCR检测。对于确诊的结核病病例（n = 62/657；9.4%），涂片显微镜检查的敏感性为82.3%。培养和RT-PCR显示出相同的敏感性，即90.3%。涂片显微镜检查、培养和RT-PCR的特异性分别为99.7%、99.4%和98.6%。mpt64 RT-PCR对痰标本中结核分枝杆菌复合群的检测具有高敏感性和特异性。该技术可应用于没有可用的结核病快速检测方法的实验室，能够在长达5小时内完成结核病诊断。
