High-resolution melting analysis for molecular detection of multidrug resistance tuberculosis in Peruvian isolates.
The emergence of multidrug-resistant strains is a major health problem especially for countries with high TB incidence such as Peru. In this study, we evaluated High Resolution Melting (HRM) assay in Peruvian isolates for the detection of mutations within rpoB, katG genes and promoter region inhA to determine isoniazid and rifampicin resistance in Mycobacterium tuberculosis (Mtb).



DNA samples extracted from a total of 167 clinical isolates of Mtb, 89 drug-sensitive and 78 multidrug-resistant, were blindly analyzed by HRM analysis and verified by DNA sequencing.



The HRM analysis generated patterns that were specific to distinguish between sensitive and resistance isolates. The sensitivity and specificity of the HRM assays in comparison with drug susceptibility testing (DST) for detection of rifampicin resistance were 98.7 % and 97.5 %, and for isoniazid resistance were 98.7 % and 100 %.



This study suggests that HRM Analysis could help with rapid diagnosis of MDR-TB cases in Peru.
Laboratorio de Biotecnología y Biología Molecular, Instituto Nacional de Salud, Lima, Peru.
Marco
M
Galarza
Laboratorio de Biotecnología y Biología Molecular, Instituto Nacional de Salud, Lima, Peru.
Manuel
M
Fasabi
Laboratorio de Biotecnología y Biología Molecular, Instituto Nacional de Salud, Lima, Peru.
Kelly S
KS
Levano
Laboratorio de Referencia Regional de Tuberculosis y Microbiología, DIRESA Callao, Lima, Peru.
Edith
E
Castillo
Laboratorio de Referencia Regional de Tuberculosis y Microbiología, DIRESA Callao, Lima, Peru.
Nadia
N
Barreda
Laboratorio de Referencia Regional de Tuberculosis y Microbiología, DIRESA Callao, Lima, Peru.
Mitzi
M
Rodriguez
Laboratorio de Biotecnología y Biología Molecular, Instituto Nacional de Salud, Lima, Peru. hguio@ins.gob.pe.
Heinner
H
Guio
D000995:Antitubercular Agents / Q000494:pharmacology* / Q000627:therapeutic use; D006801:Humans; D007538:Isoniazid / Q000494:pharmacology* / Q000627:therapeutic use; D008826:Microbial Sensitivity Tests; D009169:Mycobacterium tuberculosis / Q000187:drug effects / Q000235:genetics / Q000302:isolation & purification*; D010568:Peru / Q000453:epidemiology; D012293:Rifampin / Q000494:pharmacology* / Q000627:therapeutic use; D012680:Sensitivity and Specificity; D018088:Tuberculosis, Multidrug-Resistant / Q000175:diagnosis* / Q000453:epidemiology / Q000382:microbiology
D000995:Antitubercular Agents; D007538:Isoniazid; D012293:Rifampin
High Resolution Melting Analysis; Multi-drug resistance tuberculosis; Tuberculosis