Abstract
Drug-resistant tuberculosis (TB) is a global crisis and a threat to health security. Since conventional drug susceptibility testing (DST) takes several weeks, we herein described a molecular assay to rapidly identify multidrug-resistant (MDR) and extensively drug-resistant (XDR) and reveal transmission associated-mutations of Mycobacterium tuberculosis complex (MTBC) isolates in 6 to 7 hours. An array was designed with 12 pairs of primers and 60 single nucleotide polymorphisms of 9 genes: rpoB, katG, inhA, ahpC, embB, rpsL, gyrA, rrs and eis. We assessed the performance of the array using 176 clinical MTBC isolates. The results of culture-based DST were used as the gold standard, the GenoType MTBDRplus and MTBDRsl tests were used for parallel comparison, and gene sequencing was performed to resolve the discordance. The sensitivities and specificities of the array are comparable to those of the MTBDRplus test for resistance to isoniazid (INH) (100.0%, 96.7%) and rifampicin (RIF) (99.4%, 96.7%) and of the MTBDRsl test for resistance to fluoroquinolones (FQs) (100%, 100%) and second-line injectable drugs (SLIDs) (98.3%, 100%). The sensitivities of the array for detecting resistance to ethambutol and streptomycin were 79.3% and 64.9%, respectively. The array has potential as a powerful tool for clinical diagnosis and epidemiological investigations.
Highlights
Drug-resistant tuberculosis (TB) is a public health concern and a threat to global TB control programs
We report the performance of the new array and compare it with the performance of the GenoType MTBDRplus and GenoType MTBDRsl tests while using the conventional culture-based drug susceptibility testing (DST) as a reference test
Because of the urgent need to rapidly and accurately diagnose drug-resistant TB, the World Health Organization (WHO) has recommended the GenoType MTBDRplus and GenoType MTBDRsl tests for detecting resistance to first- and second-line drugs[1]
Summary
Drug-resistant tuberculosis (TB) is a public health concern and a threat to global TB control programs. Array-based analysis of MTBC has been shown to provide important information on the mechanism of drug resistance, the identity of the species or strain, and gene expression[9]. Commercial microarrays such as the BluePoint MycoID array[10] are capable of identifying species of MTBC and non-tuberculous mycobacteria (NTM), whereas the DR. We developed and commercialized a nylon membrane array (BluePoint MtbDR, Bio Concept Inc., Taichung, Taiwan) for detecting gene mutations conferring resistance to RIF and INH in MTBC25. We designed a novel array that can detect resistance to 8 anti-TB drugs simultaneously by including probes that can identify gene mutations conferring RIF, INH, EMB, SM, FQ, and SLIDs resistance. We report the performance of the new array and compare it with the performance of the GenoType MTBDRplus and GenoType MTBDRsl tests while using the conventional culture-based DST as a reference test
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