High throughput PCR assays for diagnosing tuberculosis

项目总结 低菌群临床标本中结核分枝杆菌的快速检测 负载对诊断实验室来说是一个挑战。基于核酸的方法，如聚合酶链式反应 由于它们的速度和高灵敏度，在这一领域有很大的前景。DevR-和 印度Pis的实验室开发了基于hupB的聚合酶链式反应分析，显示出高水平的 人类肺结核病和肺外结核病诊断的准确性 以及疑似患有结核病的受试者的牛样本。HupB试验 区别牛分枝杆菌和结核分枝杆菌，两者都是 MTB综合体。在这个项目中，提出了优化分子信标和/或引物的设计 用于以可视格式(在美国PI中)可靠地检测DevR和hupB PCR产物 实验室)。与稳健的通用样品处理(USP)技术相结合 由印度派(Jaya S.Tyagi)开发的PCR有望成为一种用户友好的测试 在所有类型的临床标本中诊断结核病。将对标准化化验结果进行测试 在印度PI的实验室中对疑似人类结核病的临床样本进行了分析 病人(肺结核病和肺外结核病)和来自经处理的农场动物 USP技术。预计这项研究的结果将得到普遍认同 适用、可靠、用户友好和高通量的诊断和/筛查分析 人体标本中流行的结核分枝杆菌复合体病原体的鉴定 和牛的血统。

Project summary The rapid detection of Mycobacterium tuberculosis in clinical samples with a low bacterial load is a challenge to the diagnostic laboratory. Nucleic acid-based approaches such as PCR offer much promise in this area owing to their speed and their high sensitivity. devR- and hupB-based PCR assays were developed in the Indian Pis' laboratories that show high diagnostic accuracy in pulmonary and extrapulmonary tuberculosis disease settings in human and bovine samples from subjects suspected to suffer from tuberculosis. The hupB assay distinguishes M. bovis from M. tuberculosis both of which are closely related members of the MTB complex. In this project it is proposed to optimise molecular beacon and / primer design for the reliable detection ofdevR and hupB PCR products in a visual format (in US Pi's laboratory). Combined with the robust Universal Sample Processing (USP) technology developed by Indian PI (Jaya S. Tyagi), PCR is poised to become a user-friendly test for diagnosing tuberculosis in all types of clinical samples. The standardized assays will be tested in the Indian Pi's laboratories on clinical samples obtained from suspected human tuberculosis patients (pulmonary & extra-pulmonary tuberculosis) and from farm animals processed by USP technology. It is expected that the results of this study will establish universally applicable, robust, user-friendly and high throughput diagnostic and / screening assays for identification of Mycobacterial pathogens of the MTB complex prevalent in samples of human and bovine origin.