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    • 2. 发明申请
    • COMPOSITIONS AND METHODS FOR A MYCOBACTERIUM TUBERCULOSIS DRUG SUSCEPTIBILITY TEST
    • MYCOBACTERIUM TUBCCOSOS药物耐药性测试的组合物和方法
    • WO2012158502A2
    • 2012-11-22
    • PCT/US2012037472
    • 2012-05-11
    • UNIV VIRGINIA PATENT FOUNDHOUPT ERIC RKELLY KIMBERLY APHOLWAT SUPORN
    • HOUPT ERIC RKELLY KIMBERLY APHOLWAT SUPORN
    • C12Q1/686A61K31/133A61K31/4409A61K31/496A61K31/506A61K31/7036C12Q1/18C12Q1/689C12Q1/701C12Q2600/136G01N2333/35
    • The present application discloses rapid Mycobacterium tuberculosis drug susceptibility utilizing real-time PCR of mycobacteriophage D29 DNA. One protocol involves culturing Tb isolates for 48 hours with and without drugs at critical concentrations, followed by incubation with 103 pfu/ml of D29 mycobacteriophage for 24 hours and then real-time PCR. Many drugs can be incubated instantly with Tb and phage. The change in phage DNA real-time PCR cycle threshold (Ct) between control Tb and Tb treated with drugs was calculated and correlated with conventional agar proportion drug susceptibility results. Specifically, 9 susceptible clinical isolates, 22 MDR, and 1 XDR Tb strains were used and Ct control - Ct drug cutoffs of between +0.3 and -6.0 yielded 422/429 (98%) accurate results for the drugs tested. The Ct values correlated with isolate minimal inhibitory concentration (MIC) for most agents. This D29 qPCR assay offers a rapid, accurate, 1-3 day phenotypic drug susceptibility test.
    • 本申请公开了利用分枝杆菌噬菌体D29DNA的实时PCR的快速结核分枝杆菌药物敏感性。 一个方案包括在临界浓度下用和不用药物培养Tb分离物48小时,然后与103pfu / ml的D29分枝杆菌噬菌体孵育24小时,然后进行实时PCR。 许多药物可以用Tb和噬菌体立即孵育。 计算用药物处理的对照Tb和Tb之间的噬菌体DNA实时PCR循环阈值(Ct)的变化,并与常规琼脂比例药物敏感性结果相关。 具体来说,使用了9种敏感性临床分离株,22种MDR和1种XDR Tb菌株,对于所测试的药物,Ct对照 - Ct药物截止值在+ 0.3和-6.0之间产生了422/429(98%)的准确结果。 Ct值与大多数试剂的分离最小抑制浓度(MIC)相关。 这种D29 qPCR测定提供了快速,准确的1-3天表型药物敏感性测试。