title: Multi-drug resistant Mycobacterium tuberculosis complex genetic diversity and clues on recent transmission in Punjab, Pakistan.
abstract: Multi-Drug Resistant Tuberculosis (MDR-TB), i.e. bacilli resistant to rifampicin (RIF) and isoniazid (INH), is a major Public Health concern in Pakistan according to WHO estimates (3.5% and 32% of new and retreated cases, respectively). Previous Pakistanis reports identified a correlation between being MDR and belonging to Beijing or EAI lineages in one study, and belonging to "H4"-Ural Euro-American sublineage in another study. In addition, MDR-TB transmission was suspected in Karachi. We tested MDR characteristics on a Punjab sample of 278 clinical isolates (without selection for Multi-Drug Resistance) including new and retreated cases collected from 2008 to 2012. All samples were characterized by a new, microbead-based method named "TB-SPRINT" (molecular diagnostic including spoligotype identification, and genetic resistance determinants to first-line anti-TB drugs RIF and INH). Isolates from 2011 to 2012 (n=100) were further analyzed using 24-loci MIRU-VNTR. We detected 8.7% MDR isolates (CI95%=[5.0; 12.5]), mainly among CAS lineage that predominates in this central-East region of Pakistan. Out of 20 MDR-TB cases, 12 different TB-SPRINT profiles were identified, limiting the suspicion of MDR-TB transmission. 24 MIRU-VNTR confirmed the unrelatedness of isolates with different TB-SPRINT profiles and discriminated 3 isolates with identical TB-SPRINT profiles. In conclusion, our study did not confirm any of the correlations between Multi-Drug Resistance and lineage or sublineage in Punjab, Pakistan. MDR-TB isolates were diverse indicating that transmission is not pervasive. TB-SPRINT proved useful as a first step for detecting MDR-TB likely transmission events, before more extensive genotyping such as 15 or 24 MIRU-VNTR and thorough epidemiological investigation.
authors: Health Biotechnology Division, National Institute for Biotechnology and Genetic Engineering (NIBGE), P.O. Box# 577, Jhang Road, Faisalabad, Pakistan; Pakistan Institute of Engineering and Applied Sciences (PIEAS), P.O. Nilore, Islamabad, Pakistan.
authors: Memona
authors: M
authors: Yasmin
authors: Institut de Génétique et Microbiologie, UMR8621, CNRS-Université Paris-Sud, Campus d'Orsay, F-91405 Orsay-Cedex, France; Centre MURAZ, Bobo-Dioulasso, Burkina Faso.
authors: Michel K
authors: MK
authors: Gomgnimbou
authors: Health Biotechnology Division, National Institute for Biotechnology and Genetic Engineering (NIBGE), P.O. Box# 577, Jhang Road, Faisalabad, Pakistan; Pakistan Institute of Engineering and Applied Sciences (PIEAS), P.O. Nilore, Islamabad, Pakistan.
authors: Rubina T
authors: RT
authors: Siddiqui
authors: Institut de Génétique et Microbiologie, UMR8621, CNRS-Université Paris-Sud, Campus d'Orsay, F-91405 Orsay-Cedex, France.
authors: Guislaine
authors: G
authors: Refrégier
authors: Institut de Génétique et Microbiologie, UMR8621, CNRS-Université Paris-Sud, Campus d'Orsay, F-91405 Orsay-Cedex, France. Electronic address: christophe.sola@u-psud.fr.
authors: Christophe
authors: C
authors: Sola
mesh_terms: D000995:Antitubercular Agents / Q000494:pharmacology; D016000:Cluster Analysis; D024901:Drug Resistance, Multiple, Bacterial / Q000235:genetics*; D014644:Genetic Variation; D005838:Genotype; D005843:Geography; D006801:Humans; D008826:Microbial Sensitivity Tests; D018598:Minisatellite Repeats; D058885:Multilocus Sequence Typing; D009154:Mutation; D009169:Mycobacterium tuberculosis / Q000145:classification / Q000235:genetics*; D010154:Pakistan / Q000453:epidemiology; D018088:Tuberculosis, Multidrug-Resistant / Q000453:epidemiology / Q000382:microbiology* / Q000635:transmission*
chemical_list: D000995:Antitubercular Agents
keywords: Genotypic drug susceptibility testing; MDR-TB; Molecular epidemiology; Rifampin