The study evaluated real-time quantitative polymerase chain reaction (qPCR) and high-resolution melt-curve analysis (HRM) for simultaneous diagnosis of osteo-articular tuberculosis (OATB) and drug resistance. Two hundred and fifty synovial fluid and pus specimens (20 confirmed OATB by culture, 130 suspected OATB, and 100 controls) processed in the Department of Medical Microbiology, PGIMER were subjected to qPCR using rpoB, MPB64, and IS6110 genes. All OATB positive specimens were subjected to HRM for detecting resistance to rifampicin and isoniazid. qPCR detected 129/150 OATB cases with a sensitivity of 86% (95% for confirmed and 84.6% for suspected OATB cases) and specificity of 100%. rpoB and MPB64 genes had higher sensitivity than IS6110 (86% vs. 74.6%). HRM reported eight multidrug resistant (MDR), two mono-rifampicin, and five mono-isoniazid resistant cases, all were concordant with gene sequencing. qPCR followed by HRM analysis offer a simple, accurate, and rapid platform for simultaneous detection of OATB and MDR.

中文翻译：

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使用实时聚合酶链反应和高分辨率熔解曲线分析诊断骨关节结核和多药耐药性

该研究评估了实时定量聚合酶链反应 (qPCR) 和高分辨率熔解曲线分析 (HRM) 对骨关节结核 (OATB) 和耐药性的同时诊断。在 PGIMER 医学微生物学系处理的 250 份滑液和脓液标本（20 份经培养证实为 OATB，130 份疑似 OATB，100 份对照）使用 rpoB、MPB64和IS6110基因进行qPCR。对所有 OATB 阳性标本进行 HRM，以检测对利福平和异烟肼的耐药性。qPCR 检测到 129/150 个 OATB 病例，灵敏度为 86%（确诊为 95%，疑似 OATB 病例为 84.6%），特异性为 100%。rpoB和MPB64基因比IS6110具有更高的敏感性（86% 对 74.6%）。HRM 报告了 8 例多药耐药 (MDR)、2 例单利福平和 5 例单异烟肼耐药病例，所有病例均与基因测序一致。qPCR 后接 HRM 分析为同时检测 OATB 和 MDR 提供了一个简单、准确和快速的平台。