Rapid detection of multidrug-resistant Mycobacterium tuberculosis by multiplex allele-specific polymerase chain reaction

• Published in: The international journal of tuberculosis and lung disease Vol. 15; no. 4; pp. 496 - 501
• Main Authors: IMPERIALE, B. R, CATALDI, A. A, MORCILLO, N. S
• Format: Journal Article
• Language: English
• Published: Paris, France IUATLD 01.04.2011; International Union against Tuberculosis and Lung Disease
• Subjects: Alleles; Antitubercular Agents - pharmacology; Argentina; Bacterial diseases; Biological and medical sciences; Drug Resistance, Multiple, Bacterial; Genes, Bacterial; Human bacterial diseases; Humans; Infectious diseases; Isoniazid - pharmacology; MAS-PCR; Medical sciences; Microbial Sensitivity Tests; Multidrug-Resistant; Mutation; Mycobacterium tuberculosis; Mycobacterium tuberculosis - drug effects; Mycobacterium tuberculosis - genetics; Mycobacterium tuberculosis - isolation & purification; Pneumology; Polymerase Chain Reaction - methods; Respiratory system : syndromes and miscellaneous diseases; Rifampin - pharmacology; Tuberculosis; Tuberculosis and atypical mycobacterial infections; Argentina; MAS-PCR; Respiratory disease; Mycobacterial infection; multidrug-resistant; Infection; Mass; Polymerase chain reaction; Allele; Rapid technique; Tuberculosis; Mycobacterium tuberculosis; Mycobacteriales; Multiple resistance; Bacteriosis; Mycobacteriaceae; Bacteria; Tumor; Actinomycetes; Molecular biology; Pneumology
• ISSN: 1027-3719; 1815-7920
• DOI: 10.5588/ijtld.10.0397

SETTING: Dr Cetrangolo Hospital, Buenos Aires Province, Argentina.OBJECTIVE: To evaluate a multiplex allele-specific polymerase chain reaction (MAS-PCR) to detect multidrug-resistant tuberculosis (MDR-TB) clinical isolates and to describe the main mutations conferring resistance to isoniazid (INH) and rifampicin (RMP).DESIGN: Drug-resistant Mycobacterium tuberculosis clinical isolates were tested to detect mutations using MAS-PCR. The genes involved were katG, inhA promoter and rpoB.RESULTS: Among 193 clinical isolates included in the study, 52.6% of the INH-resistant isolates presented a mutation in the katG (315) gene, 28.1% in the inhAP (−15) and 3.0% in both. For the rpoB gene, 60% of the RMP-resistant isolates showed a mutation in codon 531, 17.5% in 526 and 2.5% in 516. Results were compared with those obtained by sequencing, and 100% concordance was obtained for the detection of the mutation in katG (315), 94.1% for inhAP (−15), and 97.8% for rpoB. The global concordance between both methods was 98%.CONCLUSIONS: The MAS-PCR system allowed the simultaneous and rapid detection of approximately 80.0% of the drug-resistant clinical isolates. This method could be used as a rapid and simple screening tool to detect drug-resistant TB in clinical practice.