Diagnosing osteo-articular tuberculosis and multidrug resistance using real-time polymerase chain reaction and high-resolution melt-curve analysis

Sharma, Kusum; Sharma, Megha; Sharma, Aman; Dhillon, Mandeep Singh

Affiliations

¹Department of Medical Microbiology, Postgraduate Institute of Medical Education and Research (PGIMER), Chandigarh, India

²Department of Microbiology, All India Institute of Medical Sciences (AIIMS), Bilaspur, Himachal Pradesh, India

³Department of Internal Medicine, Postgraduate Institute of Medical Education and Research (PGIMER), Chandigarh, India

⁴Department of Orthopaedics, Postgraduate Institute of Medical Education and Research (PGIMER), Chandigarh, India

Abstract and keywords

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.

Keywords: melt-curve analysis; multidrug resistant tuberculosis; nucleic acid amplification test; osteoarticular tuberculosis; rpoB gene sequencing

Links

DOI: 10.1002/jor.25410

PubMed: 35780389

WoS: 000824605800001

History

Received: 2022-05-12

Revised: 2022-06-21

Accepted: 2022-06-29