Affiliation:
1. Centre of Infection and Department of Microbiology, Queen Mary Hospital, The University of Hong Kong
2. Tuberculosis and Chest Service, Department of Health
3. Tuberculosis and Chest Unit, Grantham Hospital, Hong Kong Special Administrative Region, People’s Republic of China
Abstract
ABSTRACT
This study evaluated the feasibility of a molecular strategy based on identification of
Mycobacterium tuberculosis
by IS
6110
PCR or Cobas Amplicor PCR, and
rpoB
PCR-DNA sequencing of the 81-bp rifampin resistance determining region (RRDR) for direct detection of rifampin resistance in respiratory specimens. A collection of 2,138 respiratory specimens and 352 nonduplicate
M. tuberculosis
isolates (including 233 isolates from the evaluated respiratory specimens and an additional collection of 119 stored isolates) from Southern China was investigated. Using culture as the reference test, the overall diagnostic sensitivities of an acid-fast bacillus (AFB) smear, Cobas Amplicor PCR, IS
6110
PCR were 54.5% (156 of 286), 86.7% (248 of 286), and 89.2% (255 of 286), respectively. The sensitivities of the
rpoB
PCR for the specimens with positive AFB smears and with positive PCR results in the IS
6110
PCR and/or Cobas Amplicor PCR were 100% (156 of 156) and 92.3% (239 of 259), respectively. Of the 352 nonduplicate
M. tuberculosis
isolates, the agar proportion method for rifampin reported 39 resistant strains. Full agreement (352 of 352) was found with the agar proportion method and the genotype inferred from the
rpoB
DNA sequencing data for rifampin. Thirty-nine mutations of nine distinct kinds, eight point mutations, and one deletion within the RRDR were found in the 39 resistant strains. For the direct DNA sequencing performed on
rpoB
PCR-positive respiratory specimens, the concordance with the agar proportion method and the subsequent PCR-sequencing for the culture isolate was 100%. This strategy has potential application for direct and rapid diagnosis of rifampin-resistant
M. tuberculosis
in IS
6110
PCR or Cobas Amplicor PCR-positive respiratory specimens.
Publisher
American Society for Microbiology
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