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Abstract
Background
GenoType® MTBDRplus assay (Hain Lifescience, Germany) enables detection of the mutations prevalent in rpoB, katG, and inhA genes and identification of Mycobacterium tuberculosis complex (MTB). We evaluated the performance of the MTBDRplus assay in detecting multidrug resistant M. tuberculosis in sputum specimens by directly comparing it to the performance of conventional drug susceptibility testing (DST) with M. tuberculosis culture isolates.
Methods
From December 2007 to July 2008, 40 patients with acid-fast bacilli (AFB) smear-positive and AFB culture-positive sputa, including 19 patients with rifampin (RIF)- or isoniazid (INH)-resistant MTB isolates, were enrolled. The MTBDRplus assay was performed using DNA extracted from respiratory specimens. DST of the culture isolates was performed using an absolute concentration method.
Results
The result of the AFB smear test was ±1 for 7 specimens, +1 for 8 specimens, +2 for 9 specimens, +3 for 9 specimens, and +4 for 7 specimens. The MTBDRplus assay revealed that 37 of the 40 specimens were positive for an MTB-specific band, 12 specimens were RIF-resistant, and 16 specimens were INH-resistant. The rpoB S531L mutation was detected in 58.3% of the RIF-resistant specimens, and the katG S315T1 and inhA C15T mutations were detected in 56.3% and 31.3% of the INH-resistant specimens, respectively. Compared to the sensitivity and specificity of DST, both sensitivity and specificity of MTBDRplus assay for RIF resistance were 100%, and the corresponding values for INH resistance were 82.4% and 90.0%. Discrepant MTBDRplus assay and DST results were obtained in 3 INH-resistant isolates without mutation and 2 INH-susceptible isolates with katG S315T1 and inhA C15T mutations.
Figures and Tables
Table 1
Results of the GenoType MTBDRplus assay of the specimens showing any resistant genotypes and conventional drug sensitivity testing of their isolates
Abbreviations: AFB, acid-fast bacilli; RIF, rifampin; INH, isoniazid; DST, drug susceptibility testing; MUT, mutation; WT, wild type; S, sensitive; R, resistance; rpoB WT3, 513-516 codon; rpoB WT7 526-529 codon; rpoB MUT2A, H526Y; rpoB MUT3, S531L; katG MUT1, S315T1; inhA WT1, -15 and -16; inhA WT2, -8; inhA MUT1, C15T; inhA MUT3A, T8C.
Table 2
Comparison of the GenoType MTBDRplus assay and conventional drug susceptibility test results of their isolates in 37 specimens
Table 3
Sensitivity, specificity, positive predictive value and negative predictive value of the GenoType MTBDRplus assay
Table 4
Confirmatory susceptibility testing results and previous anti-tuberculosis medication of the 5 patients showing discrepant results between GenoType MTBDRplus and conventional drug susceptibility test of the culture isolates
*These tests were performed with Mycobacterium tuberculosis culture isolates; †low-R, resistant to INH at 0.2 µg/mL but susceptible to INH at 1.0 µg/mL; ‡high-R, resistant to INH at 1.0 µg/mL.
Abbreviations: TB, tuberculosis; DST, drug sensitivity test; INH, isoniazid; RIF, rifampin; S, sensitive; R, resistance; WT, wild type; NT, not tested.
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