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1.
Indian J Med Res ; 143(3): 341-7, 2016 Mar.
Article in English | MEDLINE | ID: mdl-27241648

ABSTRACT

BACKGROUND & OBJECTIVES: Pre-extensively drug resistant (pre-XDR) and extensively drug resistant tuberculosis (XDR-TB) have been areas of growing concern, and are posing threat to global efforts of TB control. The present study was planned to study the presence of pre-XDR and XDR Mycobacterium tuberculosis and their genotypes in clinical isolates obtained from previously treated cases of pulmonary TB. METHODS: A total of 219 isolates obtained from previously treated cases of pulmonary TB were subjected to first-line (streptomycin, isoniazid, rifampicin and ethambutol) and second-line (ofloxacin, kanamycin, capreomycin and amikacin) drug susceptibility testing on solid Lowenstein-Jensen medium by proportion method. Genotyping was done for pre-XDR and XDR-TB isolates using 12 loci Mycobacterial Interspersed Repetitive Units-Variable Number Tandem Repeats (MIRU-VNTR). RESULTS: Multi-drug resistance was observed in 39.7 per cent (87/219) isolates. pre-XDR and XDR M. tuberculosis isolates amongst 87 multi-drug resistant (MDR) TB isolates were 43 (49.4%) and 10 (11.4%), respectively. Two most dominant genotypes among pre-XDR and XDR M. tuberculosis isolates were Beijing and Delhi/CAS types. INTERPRETATION & CONCLUSIONS: Resistance to second-line anti-tubercular drugs should be routinely assessed in areas endemic for TB. Similar genotype patterns were seen in pre-XDR and XDR-TB isolates. Beijing and Delhi/CAS were predominant genotypes.


Subject(s)
Drug Resistance, Multiple, Bacterial , Extensively Drug-Resistant Tuberculosis/microbiology , Mycobacterium tuberculosis/genetics , Tuberculosis, Multidrug-Resistant/microbiology , Extensively Drug-Resistant Tuberculosis/epidemiology , Extensively Drug-Resistant Tuberculosis/genetics , Genotype , Humans , India/epidemiology , Isoniazid/therapeutic use , Kanamycin/therapeutic use , Microbial Sensitivity Tests , Mycobacterium tuberculosis/drug effects , Mycobacterium tuberculosis/pathogenicity , Ofloxacin/therapeutic use , Rifampin/therapeutic use , Tuberculosis, Multidrug-Resistant/epidemiology
2.
J Antibiot (Tokyo) ; 68(1): 63-6, 2015 Jan.
Article in English | MEDLINE | ID: mdl-25052485

ABSTRACT

The study was done to know the prevalent mutations of gyrA and gyrB genes, and their significance with drug resistance in clinical isolates of Mycobacterium tuberculosis. A total of 100 ofloxacin- (OFX) resistant and 100 OFX-sensitive isolates of M. tuberculosis were consecutively selected from routine Tuberculosis laboratory. Drug resistance pattern of these isolates was recorded. MIC of OFX was tested in all these isolates by absolute concentration method. Quinolone resistance determining region (QRDR) of gyrA and gyrB genes of 320 and 428 bp, respectively, were amplified and sequenced. Sequencing data were analyzed by BLAST on NCBI with reference strain H37Rv. Of 100 OFX-sensitive isolates, 30 were pansusceptible, 28 were monoresistant, 10 were polyresistant and 32 were multidrug resistant (MDR). Among 100 OFX-resistant isolates, 19 were OFX monoresistant, 16 were polyresistant and 65 were MDR. Mutations in gyrA and gyrB genes were observed in 79% and 5% of OFX-resistant isolates, respectively. Most prevalent mutation was found at codon 94 in QRDR of gyrA gene. Double mutations found in gyrA gene and in both gyrA and gyrB genes signifies higher levels of OFX resistance. In one isolate, a substitution at codon 592 (Pro592Ser) was found as a novel mutation outside the QRDR of gyrB gene. Our findings support previous studies that the OFX resistance to M. tuberculosis is associated with mutations in the QRDR of gyrA gene; however, the level of OFX resistance may not be predicted based on the mutation patterns in the gyrA gene.


Subject(s)
Antitubercular Agents/pharmacology , DNA Gyrase/genetics , Drug Resistance, Bacterial/genetics , Mycobacterium tuberculosis/drug effects , Ofloxacin/pharmacology , Base Sequence , Codon , Drug Resistance, Multiple, Bacterial/genetics , Fluoroquinolones/pharmacology , Humans , Microbial Sensitivity Tests , Mutation , Mycobacterium tuberculosis/genetics , Mycobacterium tuberculosis/isolation & purification , Prevalence , Tuberculosis/microbiology
4.
Tuberculosis (Edinb) ; 92(5): 404-6, 2012 Sep.
Article in English | MEDLINE | ID: mdl-22789499

ABSTRACT

Resistance to second line anti tubercular drugs is a cause of serious concern. The present study reports the prevalence of Ofloxacin (OFX) and Kanamycin (KM) resistance in Mycobacterium tuberculosis isolates from cases of pulmonary tuberculosis, who received anti tubercular treatment for >4 weeks in past. Of total 438 enrolled patients, 361 were culture positive for M. tuberculosis, of which, 95 (26.3%) were OFX resistant & 49 (13.5%) were KM resistant. Total 130 isolates were Multidrug resistant, of which, 55 (42.3%) were resistant to either OFX or KM (Pre-XDR) & 11 (8.5%) were resistant to both KM & OFX (XDR). Resistance to quinolones & aminoglycosides should be routinely assessed in areas endemic for tuberculosis.


Subject(s)
Antitubercular Agents/pharmacology , Kanamycin Resistance , Kanamycin/pharmacology , Mycobacterium tuberculosis/isolation & purification , Ofloxacin/pharmacology , Tuberculosis, Multidrug-Resistant/epidemiology , Antitubercular Agents/administration & dosage , Drug Resistance, Bacterial , Female , Humans , India/epidemiology , Kanamycin/administration & dosage , Male , Microbial Sensitivity Tests , Mycobacterium tuberculosis/drug effects , Ofloxacin/administration & dosage , Population Surveillance , Prevalence , Treatment Outcome , Tuberculosis, Multidrug-Resistant/drug therapy
5.
J Microbiol Methods ; 88(1): 122-6, 2012 Jan.
Article in English | MEDLINE | ID: mdl-22133919

ABSTRACT

BACKGROUND: Drug resistant tuberculosis (TB) is a growing concern worldwide. Early detection of multidrug-resistant Mycobacterium tuberculosis is of primary importance for both patient management and infection control. Optimal method for identifying drug-resistant M. tuberculosis in a timely and affordable way in resource-limited settings is not yet available. AIM: This study evaluated; nitrate reductase assay (NRA), resazurin microtiter assay (REMA) and microscopic observation drug susceptibility assay (MODS) against the conventional 1% proportion method (PM) for the detection of resistance to first line antitubercular drugs, in M. tuberculosis clinical isolates. METHODS: A total of one hundred and five clinical isolates of M. tuberculosis; 50 pan sensitive and 55 pan resistant were tested with NRA, REMA and MODS. The 1% proportion method on Lowenstein-Jensen medium was used as reference test. RESULTS: Of all three methods which were tested NRA was found to be most sensitive and specific. Sensitivity for rifampicin resistance detection was 100%, 94.55% and 92.73% by NRA, REMA and MODS respectively. NRA and REMA were found to be 100% specific, while the MODS was 98% specific for detection of rifampicin resistance. Test results with all these methods were obtained within 8-14 days. CONCLUSION: Rapid non-conventional and inexpensive methods may serve as a replacement for 1% proportion method in resource limited settings.


Subject(s)
Antitubercular Agents/pharmacology , Microbial Sensitivity Tests/methods , Microscopy/methods , Mycobacterium tuberculosis/drug effects , Nitrate Reductase/analysis , Tuberculosis/microbiology , Bacterial Proteins/analysis , Bacterial Proteins/metabolism , Humans , Mycobacterium tuberculosis/chemistry , Mycobacterium tuberculosis/cytology , Mycobacterium tuberculosis/metabolism , Nitrate Reductase/metabolism , Nitrates/metabolism , Oxazines/analysis , Sensitivity and Specificity , Xanthenes/analysis
6.
J Biosci ; 33(4): 605-16, 2008 Nov.
Article in English | MEDLINE | ID: mdl-19208985

ABSTRACT

Drug resistant tuberculosis is a man made problem. While tuberculosis is hundred percent curable, multidrug resistant tuberculosis (MDR-TB) is difficult to treat. Inadequate and incomplete treatment and poor treatment adherence has led to a newer form of drug resistance known as extensively drug resistant tuberculosis (XDR-TB). XDR-TB is defined as tuberculosis caused by Mycobacterium tuberculosis strain, which is resistant to at least rifampicin and isoniazid among the first line anti tubercular drugs (MDR-TB) in addition to resistance to any fluroquinolones and at least one of three injectable second line anti tubercular drugs i.e. amikacin, kanamycin and/or capreomycin. Mismanagement of tuberculosis paves the way to drug resistant tuberculosis. Emergence of XDR-TB is reported world wide. Reported prevalence rates of XDR-TB of total MDR cases are; 6.6% overall worldwide, 6.5% in industrialized countries, 13.6% in Russia and Eastern Europe, 1.5% in Asia, 0.6% in Africa and Middle East and 15.4% in Republic of Korea. Better management and control of tuberculosis specially drug resistant TB by experienced and qualified doctors, access to standard microbiology laboratory,co-morbitidy of HIV and tuberculosis,new anti-TB drug regimens, better diagnostic tests,international standards for second line drugs (SLD)-susceptibility testing,invention of newer anti- tubercular molecules and vaccines and knowing the real magnitude of XDR-TB are some of the important issues to be addressed for effective prevention and management of XDR-TB.


Subject(s)
Antibiotics, Antitubercular/pharmacology , Drug Resistance, Multiple, Bacterial , Mycobacterium tuberculosis/drug effects , Tuberculosis/microbiology , Acquired Immunodeficiency Syndrome/complications , Antibiotics, Antitubercular/therapeutic use , HIV Infections/complications , Humans , Microbial Sensitivity Tests , Mycobacterium tuberculosis/genetics , Treatment Outcome , Tuberculosis/complications , Tuberculosis/drug therapy , Tuberculosis/prevention & control
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