Rapid detection of multidrug-resistant Mycobacterium tuberculosis by multiplex allele-specific polymerase chain reaction
Abstract: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.
Document Type: Regular Paper
Affiliations: 1: Reference Laboratory, Tuberculosis Control Program of Buenos Aires Province, Buenos Aires, Argentina 2: Biotechnology Institute, National Institute of Agricultural Technology, Buenos Aires, Argentina
Publication date: 2011-04-01
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