| Autor: |
Chakravorty, Soumitesh, Lee, Jong Seok, Cho, Eun Jin, Roh, Sandy S., Smith, Laura E., Lee, Jiim, Kim, Cheon Tae, Via, Laura E., Cho, Sang-Nae, Barry, Clifton E., Alland, David |
| Zdroj: |
Journal of Clinical Microbiology; November 2014, Vol. 53 Issue: 1 p43-51, 9p |
| Abstrakt: |
ABSTRACTResistance to amikacin (AMK) and kanamycin (KAN) in clinical Mycobacterium tuberculosisstrains is largely determined by specific mutations in the rrsgene and eisgene promoter. We developed a rapid, multiplexed sloppy molecular beacon (SMB) assay to identify these mutations and then evaluated assay performance on 603 clinical M. tuberculosisDNA samples collected in South Korea. Assay performance was compared to gold-standard phenotypic drug susceptibility tests, including Lowenstein-Jensen (LJ) absolute concentration, mycobacterial growth indicator tubes (MGIT), and TREK Sensititre MycoTB MIC plate (MycoTB) methods. Target amplicons were also tested for mutations by Sanger sequencing. The SMB assay correctly detected 115/116 mutant and mixed sequences and 487/487 wild-type sequences (sensitivity and specificity of 99.1 and 100%, respectively). Using the LJ method as the reference, sensitivity and specificity for AMK resistance were 92.2% and 100%, respectively, and sensitivity and specificity for KAN resistance were 87.7% and 95.6%, respectively. Mutations in the rrsgene were unequivocally associated with high-level cross-resistance to AMK and KAN in all three conventional drug susceptibility testing methods. However, eispromoter mutations were associated with KAN resistance using the MGIT or MycoTB methods but not the LJ method. No testing method associated eispromoter mutations with AMK resistance. Among the discordant samples with AMK and/or KAN resistance but wild-type sequence at the target genes, we discovered four new mutations in the whiB75' untranslated region (UTR) in 6/22 samples. All six samples were resistant only to KAN, suggesting the possible role of these whiB75' UTR mutations in KAN resistance. |
| Databáze: |
Supplemental Index |
| Externí odkaz: |
|
