title: rpoB gene high-resolution melt curve analysis: a rapid approach for diagnosis and screening of drug resistance in tuberculous meningitis.
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.
authors.affiliation: Department of Medical Microbiology, Postgraduate Institute of Medical Education and Research, Chandigarh, India. Electronic address: sharmakusum9@yahoo.co.in.
authors.forename: Kusum
authors.initials: K
authors.lastname: Sharma
authors.affiliation: Department of Neurology, Postgraduate Institute of Medical Education and Research, Chandigarh, India.
authors.forename: Manish
authors.initials: M
authors.lastname: Modi
authors.affiliation: Department of Medical Microbiology, Postgraduate Institute of Medical Education and Research, Chandigarh, India.
authors.forename: Harsimran
authors.initials: H
authors.lastname: Kaur
authors.affiliation: Department of Internal Medicine, Postgraduate Institute of Medical Education and Research, Chandigarh, India.
authors.forename: Aman
authors.initials: A
authors.lastname: Sharma
authors.affiliation: Department of Medical Microbiology, Postgraduate Institute of Medical Education and Research, Chandigarh, India.
authors.forename: Pallab
authors.initials: P
authors.lastname: Ray
authors.affiliation: Department of Internal Medicine, Postgraduate Institute of Medical Education and Research, Chandigarh, India.
authors.forename: Subhash
authors.initials: S
authors.lastname: Varma
mesh_terms: D000328:Adult; D012321:DNA-Directed RNA Polymerases / Q000235:genetics*; D024881:Drug Resistance, Bacterial; D006801:Humans; D008403:Mass Screening / Q000379:methods; D009169:Mycobacterium tuberculosis / Q000145:classification / Q000187:drug effects* / Q000235:genetics / Q000302:isolation & purification*; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D013997:Time Factors; D044366:Transition Temperature; D014390:Tuberculosis, Meningeal / Q000175:diagnosis* / Q000382:microbiology
chemical_list: D012321:DNA-Directed RNA Polymerases; C105078:RNA polymerase beta subunit
keywords: Diagnosis; High resolution melt curve; Resistance; Tuberculous meningitis