Molecular detection of rifampin and isoniazid resistance to guide chronic TB patient management in Burkina Faso.
BACKGROUND
Drug-resistant tuberculosis (DR-TB) is considered a real threat to the achievement of TB control. Testing of mycobacterial culture and testing of drug susceptibility (DST) capacity are limited in resource-poor countries, therefore inadequate treatment may occur, favouring resistance development. We evaluated the molecular assay GenoType MTBDRplus (Hain Lifescience, Germany) in order to detect DR-TB directly in clinical specimens as a means of providing a more accurate management of chronic TB patients in Burkina Faso, a country with a high TB-HIV co-infection prevalence.


METHODS
Samples were collected in Burkina Faso where culture and DST are not currently available, and where chronic cases are therefore classified and treated based on clinical evaluation and sputum-smear microscopy results. One hundred and eight chronic TB patients (sputum smear-positive, after completing a re-treatment regimen for pulmonary TB under directly observed therapy) were enrolled in the study from December 2006 to October 2008. Two early morning sputum samples were collected from each patient, immediately frozen, and shipped to Italy in dry ice. Samples were decontaminated, processed for smear microscopy and DNA extraction. Culture was attempted on MGIT960 (Becton Dickinson, Cockeysville, USA) and decontaminated specimens were analyzed for the presence of mutations conferring resistance to rifampin and isoniazid by the molecular assay GenoType MTBDRplus.


RESULTS
We obtained a valid molecular test result in 60/61 smear-positive and 47/47 smear-negative patients. Among 108 chronic TB cases we identified patients who (i) harboured rifampin- and isoniazid-susceptible strains (n 24), (ii) were negative for MTB complex DNA (n 24), and (iii) had non-tuberculous mycobacteria infections (n 13). The most represented mutation conferring rifampin-resistance was the D516V substitution in the hotspot region of the rpoB gene (43.8% of cases). Other mutations recognized were the H526D (15.6%), the H526Y (15.6%), and the S531L (9.4%). All isoniazid-resistant cases (n 36) identified by the molecular assay were carrying a S315T substitution in the katG gene. In 41.7% of cases, a mutation affecting the promoter region of the inhA gene was also detected.


CONCLUSIONS
The GenoType MTBDRplus assay performed directly on sputum specimens improves the management of chronic TB cases allowing more appropriate anti-TB regimens.
Miotto
Paolo
P
Emerging Bacterial Pathogens Unit, San Raffaele Scientific Institute, Milan, Italy. miotto.paolo@hsr.it
Saleri
Nuccia
N
Dembelé
Mathurin
M
Ouedraogo
Martial
M
Badoum
Gisèle
G
Pinsi
Gabriele
G
Migliori
Giovanni B
GB
Matteelli
Alberto
A
Cirillo
Daniela M
DM
D000995:Antitubercular Agents / Q000494:pharmacology*; D002050:Burkina Faso; D004252:DNA Mutational Analysis; D004269:DNA, Bacterial / Q000235:genetics; D006801:Humans; D007538:Isoniazid / Q000494:pharmacology*; D008826:Microbial Sensitivity Tests; D009169:Mycobacterium tuberculosis / Q000187:drug effects / Q000235:genetics*; D011933:Reagent Kits, Diagnostic; D012293:Rifampin / Q000494:pharmacology*; D012680:Sensitivity and Specificity; D013183:Sputum / Q000382:microbiology; D018088:Tuberculosis, Multidrug-Resistant / Q000175:diagnosis / Q000382:microbiology*
D000995:Antitubercular Agents; D004269:DNA, Bacterial; D011933:Reagent Kits, Diagnostic; D007538:Isoniazid; D012293:Rifampin