Multiplex allele-specific polymerase chain reaction for detection of isoniazid resistance in Mycobacterium tuberculosis
Abstract:SETTING: Pham Ngoc Thach Tuberculosis Reference Hospital, Ho Chi Minh City, Viet Nam.
DESIGN: A multiplex allele-specific polymerase chain reaction (MAS-PCR) was developed to detect mutations at the two most common sites responsible for isoniazid (INH) resistance in Mycobacterium tuberculosis: katG315 and inhA-15. The MAS-PCR is able to detect rare mutations at katG315, in addition to katG S315T. Conventional phenotypic proportion drug susceptibility testing on Löwenstein-Jensen media was used as a gold standard to compare the sensitivity and specificity of the commercial MTBDRplus line-probe assay and the MAS-PCR in 100 INH-resistant and 50 INH-susceptible isolates collected consecutively at Pham Ngoc Thach Hospital reference laboratory.
RESULTS: The sensitivity and specificity on culture isolates were 90% (n = 90/100, 95%CI 0.83–0.94) and 100% (n = 50/50, 95%CI 0.93–1.0), respectively, for the MAS-PCR and the MTBDRplus assay.
CONCLUSION: The MAS-PCR described here represents an alternative method for rapid screening for INH resistance in M. tuberculosis isolates.
Document Type: Regular Paper
Affiliations: 1: University of Oxford Clinical Research Unit, Wellcome Trust Major Overseas Programme, Hospital for Tropical Diseases, Ho Chi Minh City, Viet Nam 2: Pham Ngoc Thach Hospital for Tuberculosis and Lung Diseases, Ho Chi Minh City, Viet Nam 3: Hospital of Tropical Diseases, Ho Chi Minh City, Viet Nam 4: University of Oxford Clinical Research Unit, Wellcome Trust Major Overseas Programme, Hospital for Tropical Diseases, Ho Chi Minh City, Viet Nam; Centre for Clinical Vaccinology and Tropical Medicine, Churchill Hospital, Oxford, UK
Publication date: June 1, 2011
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