Mutations in rpoB and katG genes in Mycobacterium isolates from the Southeast of Mexico.

The most frequent mutations associated with rifampin and isoniazid resistance in Mycobacterium are the substitutions at codons 531 and 315 in the rpoB and katG genes, respectively. Hence, the aim of this study was to characterize these mutations in Mycobacterium isolates from patients suspected to be infected with drug-resistant (DR) pulmonary tuberculosis (TB) in Veracruz, Mexico. Drug susceptibility testing of 25 clinical isolates revealed that five were susceptible while 20 (80%) were DR (15% of the annual prevalence for Veracruz). Of the DR isolates, 15 (75%) were resistant to rifampin, 17 (85%) to isoniazid and 15 (75%) were resistant to both drugs (MDR). Sequencing analysis performed in the isolates showed that 14 (93%) had mutations in the rpoB gene; seven of these (47%) exhibited a mutation at 531 (S-->L). Ten (58%) of the 20 resistant isolates showed mutations in katG; nine (52%) of these 10 exhibited a mutation at 315 (S-->T). In conclusion, the DR profile of the isolates suggests a significant number of different DR-TB strains with a low frequency of mutation at codons 531 and 315 in rpoB and katG, respectively. This result leads us to consider different regions of the same genes, as well as other genes for further analysis, which is important if a genetic-based diagnosis of DR-TB is to be developed for this region.

Evaluation of new external quality assessment guidelines involving random blinded rechecking of acid-fast bacilli smears in a pilot project setting in Mexico.

SETTING: Laboratories in Mexico that support the national tuberculosis (TB) control program have been involved in an acid-fast bacilli (AFB) microscopy external quality assurance program which includes rechecking 100% of smears identified as AFB-positive by the local laboratories and 10% of smears identified as AFB-negative. Very few errors have been detected in Mexico using non-random selection and unblinded rechecking of the slides. OBJECTIVE: To evaluate the results from a 1-year pilot program involving blinded rechecking of randomly selected AFB slides from local TB laboratories in two Mexican states and determine its feasibility for future implementation. DESIGN: To reduce potential bias, laboratory staff from the National TB Laboratory, Institute for Epidemiological Diagnosis and Reference (InDRE), performed quarterly statistical sampling of AFB smears and on-site evaluations in local laboratories in each state. AFB smears were rechecked at the respective state laboratories with discordant results resolved at InDRE. RESULTS: A significantly greater percentage of errors was detected on the randomly selected, blinded AFB smears than on the non-randomly selected, unblinded smears. CONCLUSION: Random blinded rechecking provides more accurate estimates of AFB microscopy results, resulting in improved diagnosis and monitoring of treatment response.

Mutations in rpoB and katG genes in Mycobacterium isolates from the Southeast of Mexico

The most frequent mutations associated with rifampin and isoniazid resistance in Mycobacterium are the substitutions at codons 531 and 315 in the rpoB and katG genes, respectively. Hence, the aim of this study was to characterize these mutations in Mycobacterium isolates from patients suspected to be infected with drug-resistant (DR) pulmonary tuberculosis (TB) in Veracruz, Mexico. Drug susceptibility testing of 25 clinical isolates revealed that five were susceptible while 20 (80 percent) were DR (15 percent of the annual prevalence for Veracruz). Of the DR isolates, 15 (75 percent) were resistant to rifampin, 17 (85 percent) to isoniazid and 15 (75 percent) were resistant to both drugs (MDR). Sequencing analysis performed in the isolates showed that 14 (93 percent) had mutations in the rpoB gene; seven of these (47 percent) exhibited a mutation at 531 (S[L). Ten (58 percent) of the 20 resistant isolates showed mutations in katG; nine (52 percent) of these 10 exhibited a mutation at 315 (S[T). In conclusion, the DR profile of the isolates suggests a significant number of different DR-TB strains with a low frequency of mutation at codons 531 and 315 in rpoB and katG, respectively. This result leads us to consider different regions of the same genes, as well as other genes for further analysis, which is important if a genetic-based diagnosis of DR-TB is to be developed for this region.