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Diagn Microbiol Infect Dis. 2015 Oct;83(2):144-9. doi: 10.1016/j.diagmicrobio.2015.06.010. Epub 2015 Jun 18.

rpoB gene high-resolution melt curve analysis: a rapid approach for diagnosis and screening of drug resistance in tuberculous meningitis.

Author information

1
Department of Medical Microbiology, Postgraduate Institute of Medical Education and Research, Chandigarh, India. Electronic address: sharmakusum9@yahoo.co.in.
2
Department of Neurology, Postgraduate Institute of Medical Education and Research, Chandigarh, India.
3
Department of Medical Microbiology, Postgraduate Institute of Medical Education and Research, Chandigarh, India.
4
Department of Internal Medicine, Postgraduate Institute of Medical Education and Research, Chandigarh, India.

Abstract

Timely and rapid diagnosis of multidrug resistance in tuberculous meningitis (TBM) is a challenge both for a microbiologist and neurologist. The present study was conducted to evaluate role of real-time polymerase chain reaction (PCR) using rpoB, IS6110, and MPB64 as targets in diagnosis of TBM in 110 patients and subsequent high-resolution melt (HRM) curve analysis of rpoB gene amplicons for screening of drug resistance. The sensitivity of smear, culture, and real-time PCR was 1.8%, 10.9%, and 83.63%, respectively. All 120 control patients showed negative results. With HRM rpoB analysis, rifampicin resistance was detected in 3 out of 110 cases of TBM (3.33%). Subsequently, results of HRM analysis were confirmed by rpoB gene sequencing, and mutations were observed at 516 (2 patients) and 531 (1 patient) codons, respectively. rpoB HRM analysis can be a promising tool for rapid diagnosis and screening of drug resistance in TBM patients in 90minutes.

KEYWORDS:

Diagnosis; High resolution melt curve; Resistance; Tuberculous meningitis

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