title: Improved rapid molecular diagnosis of multidrug-resistant tuberculosis using a new reverse hybridization assay, REBA MTB-MDR.
abstract: Rapid diagnosis of multidrug-resistant tuberculosis (MDR-TB) is essential for the prompt initiation of effective second-line therapy to improve treatment outcome and limit transmission of this obstinate disease. A variety of molecular methods that enable the rapid detection of mutations implicated in MDR-TB have been developed. The sensitivity of the methods is dependent, in principle, on the repertoire of mutations being detected, which is typically limited to mutations in the genes rpoB, katG and the promoter region of inhA. In this study, a new reverse hybridization assay, REBA MTB-MDR (M&D), that probes mutations in the oxyR-ahpC intergenic region, in addition to those in rpoB, katG and the inhA promoter region, was evaluated. A set of 240 Mycobacterium tuberculosis clinical isolates from patients receiving retreatment regimens was subjected to conventional phenotypic drug-susceptibility testing (DST) and the REBA MTB-MDR assay. The nucleotide sequences of the loci known to be involved in drug resistance were determined for comparison. In brief, the results showed that the REBA MTB-MDR assay efficiently recognized nucleotide changes in the oxyR-ahpC intergenic region as well as those in rpoB, katG and the inhA promoter region with higher sensitivity, resulting in an 81.0 % detection rate for isoniazid resistance. Inclusion of the oxyR-ahpC intergenic region in the REBA MTB-MDR assay improved the overall sensitivity of molecular DST for MDR-TB from 73.1 to 79.9 %.
authors: Department of Biomedical Laboratory Science, College of Health Sciences, Yonsei University, Wonju 220-710, Republic of Korea.
authors: Hyeeun
authors: H
authors: Bang
authors: Department of Biomedical Laboratory Science, College of Health Sciences, Yonsei University, Wonju 220-710, Republic of Korea.
authors: Sangjung
authors: S
authors: Park
authors: Department of Biomedical Laboratory Science, College of Health Sciences, Yonsei University, Wonju 220-710, Republic of Korea.
authors: Joohwan
authors: J
authors: Hwang
authors: Department of Biomedical Laboratory Science, College of Health Sciences, Yonsei University, Wonju 220-710, Republic of Korea.
authors: Hyunwoo
authors: H
authors: Jin
authors: International Tuberculosis Research Center, Changwon 631-710, Republic of Korea.
authors: Eunjin
authors: E
authors: Cho
authors: Mokpo National Hospital, Mokpo 530-828, Republic of Korea.
authors: Dae Yoon
authors: DY
authors: Kim
authors: International Tuberculosis Research Center, Changwon 631-710, Republic of Korea.
authors: Taeksun
authors: T
authors: Song
authors: Tuberculosis Research Section, Laboratory of Clinical Infectious Disease, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, MD 20852, USA.
authors: Isdore Chola
authors: IC
authors: Shamputa
authors: Tuberculosis Research Section, Laboratory of Clinical Infectious Disease, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, MD 20852, USA.
authors: Laura E
authors: LE
authors: Via
authors: Tuberculosis Research Section, Laboratory of Clinical Infectious Disease, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, MD 20852, USA.
authors: Clifton E
authors: CE
authors: Barry
authors: The Genome Research Center for Respiratory Pathogens, Yonsei University College of Medicine, Seoul 120-752, Republic of Korea.
authors: Sang-Nae
authors: SN
authors: Cho
authors: Department of Biomedical Laboratory Science, College of Health Sciences, Yonsei University, Wonju 220-710, Republic of Korea.
authors: Hyeyoung
authors: H
authors: Lee
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