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Demonstrating a Multi-drug Resistant Mycobacterium tuberculosis Amplification Microarray
Published on: April 25, 2014
Diagnosing osteo-articular tuberculosis and multidrug resistance using real-time polymerase chain reaction and
Kusum Sharma1, Megha Sharma2, Aman Sharma3
1Department of Medical Microbiology, Postgraduate Institute of Medical Education and Research (PGIMER), Chandigarh, India.
Abstract:
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.
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