Identifica los linajes de Mycobacterium tuberculosis (MTB) en la región Callao mediante el uso de la metodología MIRU-VNTR. El estudio comprende 133 muestras de DNA obtenidas a partir de aislamientos de MTB en medio sólido Löwenstein - Jensen (LJ). Utiliza 24 MIRU-VNTR para la genotipificación de MTB. Los resultados reportan un alto poder discriminatorio de este método con un HGDI de 0.995. El linaje LAM es el más prevalente (51.1 %), seguido por Haarlem (18.8 %), el linaje asiático Beijing (8.3 %), el linaje Uganda se identificó en el 6% y los linajes T y S se observaron con un 2.3 % y 0.8 %, respectivamente. Identifica como Orphans (huérfanos) un 8.3 % de las cepas analizadas. La región Callao muestra una gran diversidad de linajes; así como, la presencia de patrones huérfanos endémicos. Esta información permite reconocer los linajes que se encuentran circulando en toda la región, así mismo su posible relación con multidrogorresistencia, lo cual será de gran utilidad al sector salud, para establecer programas de control y prevención oportuna de casos de tuberculosis. / Tesis

Dissertation Submitted to the Faculty of Health Sciences, University of the Witwatersrand, Johannesburg, in the fulfillment of the requirement for the degree of Master of Science in Medicine by Research. Johannesburg, 2017 / Electron transport and respiration in Mycobacterium tuberculosis (Mtb), the causative agent of tuberculosis (TB) occurs through the use of the aa3 type cytochrome c oxidase (CcO) under normoxic conditions or cytochrome bd oxidase (CbdO) under microaerophillia. Using these oxidases, Mtb couples substrate level oxidation to generation of metabolic energy in the form of adenosine triphosphate (ATP) under aerobic conditions. The presence of the CbdO is expected to allow for growth and survival of Mtb in the oxygen restricted environment of the human TB granuloma, thus rendering it an important enzyme for further study. CbdO is comprised of two structural subunits, CydA (subunit I) and CydB (subunit II), which are encoded by the cydAB operon. Both subunits span the cytoplasmic membrane to form part of the mycobacterial electron transport chain. Notably, two other genes that are transcribed separately from the cydAB operon, the cydDC operon, have been proposed to be required for the synthesis of a functional CbdO. Based on amino acid sequence and structural predictions, the cydDC encode heterodimeric members of the ATP Binding Cassette ABC-type transporters. In other organisms, the cydDC functions to transport reducing agents to the periplasm, thus contributing to periplasmic redox homeostasis. In this study we aimed to analyze the function of the cydDC genes in Mycobacterium smegmatis. Through bioinformatics analyses, it was demonstrated that the CydDC subunits retain conserved residues associated with the ABC domain and adopt a three-dimensional fold that is similar to their counterparts in Escherichia coli. However, the published sequence of M. smegmatis suggests that cydC is a pseudogene, which was inconsistent with the demonstrated evidence of a functional CbdO in this organism. In this study, using standard DNA sequencing, it was demonstrated that the CBTBR laboratory strain of M. smegmatis does not harbor a cydC pseudogene but rather has a functional cydC gene. Next, we interrogated the function of the M. smegmatis and Mtb cydDC genes by heterologous expression in an E. coli cydD mutant. Heterologous expression of the Mtb cydDC genes restored CbdO biogenesis in the E. coli mutant. Using various microbiological approaches, we demonstrated that the mycobacterial cydDC was able to revert the stationary phase exit defect, high temperature sensitivity and increased oxidative stress susceptibility defects of the E. coli cydD mutant. Collectively, these data provided strong evidence that the mycobacterial cydDC genes encode a functional transporter that contributes to periplasmic redox homeostasis. Following this, we generated a double deletion mutant of the cydDC operon in M smegmatis. We confirmed the genetic integrity of the ΔcydDC strain by Southern Blot analysis and proved by absorption difference spectroscopy that this strain is defective in the ability to synthesize a functional CbdO, as measured by the lack of a heme d peak in membrane preparations from the mutant. In addition, the ΔcydDC mutant displayed increased sensitivity to oxidative stress and a reduced ability to exit stationary phase, phenotypic defects that were consistent with the lack of CbdO. In summary, this study provides the first evidence that loss of the M. smegmatis cydDC genes affects CbdO biogenesis. These data also confirm that the CydDC ABC-type transporter most likely transports reducing equivalents that allow for maturation of CbdO in the periplasm. Collectively, our observations advance the understanding of the mycobacterial electron transport chain and provide new evidence to assist in the development of CbdO as a TB drug target. / MT2017

Mycobacterium tuberculosis (MTB) encodes three enzymes that are dependent on vitamin B12–derived cofactors for activity, including a B12-dependent methionine synthase (MetH). Previously, work in the Molecular Mycobacteriology Research Unit (MMRU) demonstrated vitamin B12 auxotrophy in a mutant strain disrupted in the alternative, B12-independent methionine synthase, MetE. This observation established the ability of MTB to transport corrinoids despite the absence of an identifiable B12-specific transporter. In addition, it suggested that MTB does not synthesize vitamin B12 in vitro. Notably, bioinformatic analyses identified PPE2 as the only B12-related transport candidate in MTB, though as a putative B12-regulated cobalt transporter. PPE2 is unusual in possessing directly upstream of its predicted start codon one of only two B12-dependent riboswitches in the MTB genome, and it lies in a putative operon with B12 biosynthetic genes, cobU and cobQ1. In this study, the possibility that PPE2 functions in the transport of vitamin B12 or cobalt was investigated. Transcriptional and phenotypic data suggested that PPE2 was not involved in B12 transport. Instead, it was shown that cobalt can supplement the growth of an MTB metE mutant in liquid medium, strongly supporting the ability of MTB to synthesize B12 de novo. Moreover, the ability to utilise exogenous cobalt was dependent on functional PPE2, thereby establishing a role for a PPE-family member in cobalt assimilation in MTB. Vitamin B12 comprises a central corrin ring co-ordinated to 5,6-dimethylbenzimidazole (DMB) as α-axial ligand. Substituting DMB with adenine yields the alternate form, pseudo-B12. The ability of mycobacteria to utilize pseudo-B12 precursors (cobinamide and adenine) to support full function of B12-dependent metabolic pathways was evaluated. Although the pseudo-B12 precursors appeared to complement chemically the mycobacterial B12 auxotrophs, growth of the mutants on cobinamide alone complicated this interpretation. To address this limitation, DMB synthesis was targeted by disrupting the MTB bluB homologue, Rv0306. Neither site-directed mutagenesis of key Rv0306 residues, nor full-gene deletion was sufficient to eliminate growth on cobinamide. Instead, this observation highlights the need to establish biochemically the nature of the active B12 form synthesized and utilized by MTB under different conditions. In combination, the results presented here support the inferred flexibility of vitamin B12 biosynthesis in MTB, and reinforce the potential role of B12-dependent metabolism in mycobacterial pathogenesis.

Submitted by PPG Biologia Celular e Molecular (bcm@pucrs.br) on 2018-07-26T14:44:00Z No. of bitstreams: 1 JOSE_EDUARDO_SACCONI_NUNES_DIS.pdf: 1721834 bytes, checksum: 00ef2ffd5994d7fa6e05348a79aaced6 (MD5) / Approved for entry into archive by Sheila Dias (sheila.dias@pucrs.br) on 2018-07-31T14:27:33Z (GMT) No. of bitstreams: 1 JOSE_EDUARDO_SACCONI_NUNES_DIS.pdf: 1721834 bytes, checksum: 00ef2ffd5994d7fa6e05348a79aaced6 (MD5) / Made available in DSpace on 2018-07-31T14:47:08Z (GMT). No. of bitstreams: 1 JOSE_EDUARDO_SACCONI_NUNES_DIS.pdf: 1721834 bytes, checksum: 00ef2ffd5994d7fa6e05348a79aaced6 (MD5) Previous issue date: 2011-04-15 / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior - CAPES / In 2009, tuberculosis (TB) was responsible for 1.3 million deaths worldwide. The incidence rates reached 9.4 millions and the World Health Organization (WHO) estimative indicates that one third of the world population is infected by Mycobacterium tuberculosis, the main agent responsible for the disease. The lack of new drugs released on market, the long period treatment presenting side effects (causing the abandon by the patients) and the cases with HIV co-infection contributed to the appearance of multi drug resistant strains (MDR-TB) and extensively drug resistant strains (XDR-TB). It's clear, thus, that the development of new drugs to fight TB is necessary and fundamental to the success in eradicating this disease. The histidine biosynthesis pathway emerge in this context offering attractive targets, given that its present in prokaryotes, lower eukaryotes and plants, but absent in animals. The last enzyme in the route is called Histidinol Dehydrogenase and is responsible for the conversion of L-Histidinol into LHistidine. Its essentiality to the bacilli was confirmed by gene knockout, confirming its potential for the development of inhibitory compounds. In this work, a purification protocol was developed, producing the enzyme in the homogeneous form in quantities sufficient to carry its biochemical characterization. The enzyme needs a divalent metal ion in the active site to catalyze the reaction. The kinetic constants were determined, as well as the mechanism, the pH rate profiles and the interaction of its substrates and products by isothermal titration calorimetry. A tridimensional model for its structure was constructed by sequence homology, allowing the analysis of the interaction of the substrates and metal in the active site. The results obtained will allow the rational design of molecules that act as inhibitors. / Em 2009 a tuberculose (TB) foi respons?vel por 1,3 milh?es de mortes no mundo inteiro. A incid?ncia de casos chegou ao patamar de 9,4 milh?es e as estimativas da Organiza??o Mundial da Sa?de (OMS) indicam que aproximadamente 1/3 da popula??o mundial est? infectada pelo Mycobacterium tuberculosis, principal agente causador da doen?a. A falta de novas drogas no mercado, o tratamento longo e com efeitos colaterais (levando ao abandono por parte dos pacientes) e os quadros de co-infec??o com HIV tem colaborado para o surgimento de novas cepas resistentes as drogas atualmente em uso (MDR-TB e XDR-TB). Fica claro, portanto, que o desenvolvimento de novas drogas para o combate da TB ? necess?rio e fundamental para que se tenha sucesso na erradica??o desta doen?a. A via de bioss?ntese de histidina aparece nesse contexto oferecendo alvos atrativos, visto que est? presente em organismos procari?ticos, em organismos eucari?ticos inferiores e em plantas, mas ausente em animais. A ?ltima enzima pertencente ? via ? chamada de Histidinol Desidrogenase e ? respons?vel pela convers?o de L-Histidinol em L-Histidina. Sua essencialidade para a viabilidade do bacilo foi comprovada atrav?s de nocaute g?nico, confirmando sua potencialidade para o desenvolvimento de compostos inibidores de sua atividade. Neste trabalho, um protocolo depurifica??o foi desenvolvido, produzindo a enzima na forma homog?nea em quantidades suficientes para realizar a caracteriza??o bioqu?mica da mesma. A enzima necessita de um ?on met?lico divalente no s?tio para catalisar a rea??o. Suas constantes cin?ticas foram determinadas, assim como o mecanismo, os perfis de pH, e a intera??o com os substratos e produtos atrav?s de calorimetria de titula??o isot?rmica. Um modelo tridimensional da sua estrutura foi constru?do por homologia de sequ?ncia, permitindo uma an?lise da intera??o dos substratos e do metal no s?tio ativo da enzima. Os resultados obtidos permitir?o o desenho racional de mol?culas que atuem como inibidores.

Orientador: Lilian Castiglioni / Banca: Débora Aparecida Pires de Campos Zuccari / Banca: Fernando Rogério Pavan / Banca: Daisy Nakamura Sato / Banca: Margarete Gotardo de Almeida / Resumo: A tuberculose é considerada um problema prioritário de saúde pública. Há cada vez mais evidências de que, a variação das cepas de M. tuberculosis tem um papel importante no resultado da infecção por tuberculose. No presente estudo, além de descrever os aspectos sociodemográficos, clínico-epidemiológicos e bacteriológicos da tuberculose e relacionar com a distribuição de resistência aos fármacos antituberculose, também objetivou-se avaliar a diversidade dos genótipos de isolados de Mycobacterium tuberculosis utilizando as técnicas de Spoligotipyng (Spacer Oligonucleotide Typing) e MIRU-VNTR (Mycobacterial Interspersed Repetitive Units-variable-number tandem repeats). Com relação à resistência aos fármacos, as culturas de MT e os testes de sensibilidade mostram que cepas de pessoas que fizeram tratamento de TB anterior ou infectados pelo HIV apresentaram mais resistência os fármacos quando comparados aos que não apresentavam essas características (p<0,05). Os casos resistentes, em sua maioria, são de origem pulmonar e apresentam baciloscopia positiva. A análise molecular mostrou que das 377 amostras com perfis completos para 14 loci MIRU-VNTR, os loci 10, 16, 23, 26, 27, 31, 39 e 40 tiveram um alto poder discriminatório (0.6), os loci 2, ETRB e Mtub21 foram moderadamente discriminatórios (0.3) e loci 4, 20 e 24, baixo poder discriminatório (0.3). Dos 250 isolados de M. tuberculosis analisados por spoligotyping, 92 diferentes padrões foram observados e diferentes famílias foram identificadas: Haarlem (H), Latin American and Mediterraneam (LAM), T Family, undesignated (U), S lineage e X Family. Significativa variabilidade genética foi observada nos isolados de M. tuberculosis circulantes na região Noroeste Paulista quando analisados por spoligotyping e MIRU-VNTR / Abstract: Tuberculosis is considered a high priority public health problem. There is increasing evidence that the variation in Mycobacterium tuberculosis strains play an important role in the outcome of infections by tuberculosis. Apart from describing sociodemographic, clinical, epidemiological and bacteriological aspects of tuberculosis (TB) and correlating the distribution of resistance to antituberculosis drugs, one objective of this study was to evaluate the diversity of the genotypes of M. tuberculosis isolates using the Spoligotyping (spacer oligonucleotide typing) and MIRU-VNTR (Mycobacterial Interspersed Repetitive Units variable-number tandem repeats) techniques. With respect to drug resistance, cultures of M. tuberculosis and susceptibility tests showed that strains from people previously submitted to TB treatment or infected with HIV have more resistance to drugs compared to those without these characteristics (p <0.05 ). Resistant cases are mostly pulmonary M. tuberculosis, and have positive smear test results. Molecular analysis showed that of the 377 samples, 14 MIRU-VNTR loci had complete profiles; the 10, 16, 23, 26, 27, 31, 39 and 40 loci had high discriminatory power (0.6) the 2, ETRB and Mtub21 loci were moderately discriminatory (0.3) and 4, 20 and 24 loci had low discriminatory power (0.3). Ninety-two different patterns were observed in the 250 M. tuberculosis isolates analyzed by spoligotyping and different families were identified as Haarlem (H), Latin American and Mediterranean (LAM), T Family, undesignated (U), S lineage and X Family. Significant genetic variability was observed in M. tuberculosis isolates circulating in the northwestern region of Sao Paulo State when analyzed by spoligotyping and MIRU-VNTR / Doutor

Resumo: De acordo com a OMS, a tuberculose mata 5000 pessoas por dia, e se não controlada, são estimados 35 milhões de novos casos nos próximos vinte anos. A alta susceptibilidade dos infectados por HIV para a tuberculose, bem como a proliferação da tuberculose multidroga-resistente (MDR), tem criado um grande interesse mundial de expansão nos programas atuais de pesquisas sobre a tuberculose. A Purina Nucleosídeo Fosforilase do Mycobacterium tuberculosis (MtPNP) é um potencial alvo para novas drogas anti-tuberculose visto que é responsável pela síntese de novo de ribonucleotídeos de purina. A Inibição específica da PNP poderia potencialmente levar o M. tuberculosis ao estado latente. O objetivo desde trabalho é resolver a estrutura da MtPNP complexada com adenina, e analisar as interações com os ligantes. A MtPNP foi cristalizada utilizando condições experimentais descritas posteriormente, e o ligante (adenina) foi adicionado por soaking. Os dados de difração de raios X foram coletados no detector CCD utilizando fonte de radiação síncotron (Laboratório Nacional de Luz Síncrotron, LNLS, Campinas, Brazil). O programa MOSFLM foi utilizado para o processamento, e os dados foram escalonados através do programa SCALA, apresentando o grupo espacial ortorrômico P21212 (a=118,96Å, b=134,65 Å, c=44,42 Å). O cristal foi determinado utilizando o método de substituição...(Resumo completo, clicar acesso eletrônico abaixo) / Abstract: In according to the WHO, the tuberculosis kills 5000 people every day, if does not controlled, are esteemed 35 millions of new cases in next 20 years. The high susceptibility of human immunodeficiency virus infected persons to the disease and the proliferation of multidrug-resistant (MDR) strains have created a worldwide interest in expanding current programs in tuberculosis research. The Purine Nucleoside Phosphorylase from Mycobacterum tuberculosis (MtPNP) is a potential target for new drug anti-tuberculosis, whereas is responsible for de novo synthesis of purine ribonucleotides. The specific inhibition of M. tuberculosis PNP could potentially lead to the latent state of M. tuberculosis. The objective of this work is to solve the structure from MtPNP complexed with adenine, and to analyze their interactions with the ligand. MtPNP was crystallized using the experimental conditions described elsewhere, and the ligand (adenine) was added by soaking. The X-ray diffraction data were collected on a CCD detector using synchrotron radiation source (Laboratório Nacional de Luz Síncrotron, LNLS, Campinas, Brazil). It was used the MOSFLM program to processing, and the data were scaled through the program SCALA, presenting orthorhombic spatial group P21212 (a=118,96Å, b=134,65 Å, c=44,42 Å). The crystal was determined by molecular replacement methods using the program AMoRe. The final model has Rfree and Rfactor of 23.87% and 17.51% respectively, and maximum resolution of 1.86Å. It was observed a large...(Complete abstract click electronic access below) / Orientador: Fernanda Canduri / Coorientador: Walter Filgueira de Azevedo Júnior / Banca: José Roberto Ruggiero / Banca: Flávio Augusto Vicente Seixas / Mestre

Orientador: Fernando Rogério Pavan / Banca: Patricia Bento da Silva / Banca: Jean Leandro dos Santos / Resumo: A tuberculose (TB) é uma doença infecciosa que tem como principal patógeno o Mycobacterium tuberculosis e continua sendo um importante problema de saúde pública mundial, exigindo o desenvolvimento de estratégias para o seu controle. Em 2011 foram notificados 8,7 milhões de casos da doença no mundo. Ao longo dos anos o cenário da doença não tem se mostrado otimista, devido ao aumento de número de casos de TB multi resistente a fármacos (TB-MDR) e o surgimento de cepas de resistência estendida (TB-XDR). A pesquisa de novos fármacos, em um contexto geral, apresenta-se como um enorme desafio científico para a era moderna. Neste sentido, a Química Inorgânica Medicinal tem se mostrado uma ferramenta bastante promissora. Este trabalho objetivou a caracterização da atividade anti- M. tuberculosis intra e extracelular e a citotoxicidade de 158 compostos de coordenação com metais. A citotoxicidade usando linhagens celulares de macrófago (J774A.1) e células epiteliais (VERO) também foi investigada. Diante Os resultados demonstraram que 16 compostos apresentaram uma alta seletividade, ou seja, alta atividade contra o bacilo da tuberculose e baixa citotoxicidade frente às linhagens testadas. Quatro desses 16 compostos selecionados foram analisados quanto a atividade intracelular; dos quais 2 compostos de coordenação de Co (cobalto) mostraram-se promissores quanto a esta atividade. Com base nos resultados encontrados mais estudos serão realizados a fim de garantir a eficácia e segurança desses novos compostos de coordenação candidatos à fármacos para tratamento da tuberculose / Abstract: Not available / Mestre

Em todo o mundo, menos de 5% dos doentes com tuberculose (TB), sejam casos novos ou previamente tratados, tem a avaliação dos isolados quanto ao perfil de sensibilidade aos antibioticos. No Rio Grande do Sul, estado localizado no sul do Brasil, cerca de 4700 casos novos de TB são registrados a cada ano, com uma taxa de cura de 68,9%, e uma taxa de abandono de 7,5%. A identificação rápida da resistência às drogas, em isolados clínicos de M. tuberculosis é importante para o estabelecimento de uma quimioterapia eficaz bem como para evitar a propagação de cepas resistentes. Os objetivos deste estudo foram caracterizar os pacientes de TB com maior risco de possuir TB-­‐MDR, analisando o perfil de resistência às drogas dos isolados e o perfil epidemiológico desses pacientes. Além disso utilizou-­‐se as amostras clínicas para avaliar o teste comercial (GenoType® MTBDRplus) e para desenvolver e padronizar um novo teste (Detect-­‐TBMR) para detectar as mutações mais frequentes associadas a resistência à INH e RIF. Uma proporção significativamente maior (75% versus 20%, p = 0,009) de pacientes do gênero masculino foi encontrada entre os casos resistentes às drogas do que entre os casos suscetíveis. 43,8% dos pacientes demoraram mais de 30 dias para procurar assistência médica e no grupo TB MDR, 25% dos casos não tinha sido submetido a qualquer tratamento prévio anti-­‐TB. Em nossas amostras, encontramos uma proporção de 48,3% de TB-­‐ MDR. A família T foi a família de spoligotipo mais frequente. Comparado com o método da proporções, a sensibilidade e especificidade do ensaio MTBDRplus foram 82% e 94% para a resistência à RIF, 60% e 94% para resistência à INH. Comparado com sequenciamento, a sensibilidade e especificidade do ensaio MTBDRplus foi 92% e 97% para a resistência à RIF e 100% e 100% para a resistência à INH, respectivamente. Para detectar resistência à RIF e INH, o ensaio Detect-­‐TBMDR mostrou sensibilidade e especificidade de 79,3% e 77,0% e 100% e 65%, respectivamente, em comparação com o método da proporções. Comparado com o sequenciamento, a sensibilidade e especificidade do ensaio Detect-­‐TBMDR foi de 81,2% e 94,7% e 100% e 96,2%, para detectar e resistência à RIF e INH, respectivamente. Ainda existem discordâncias entre o método das proporções e a abordagem molecular, particularmente em relação a resistência à INH. Contudo, estes métodos são muito importantes para o manejo mais rápido e correto dos pacientes, auxiliando na escolha do melhor esquema terapêutico. / In most parts of the world, less than 5% of new and previously treated tuberculosis (TB) patients are tested for multidrug resistance (MDR) TB. In Rio Grande do Sul state, the southern most Brazilian state; approximately 4700 new cases of TB are recorded each year, with a cure rate of 68.9%, and a noncompliance rate of 7.5%. Rapid identification of drug resistance in clinical isolates of Mycobacterium tuberculosis is important to facilitate rapid and adequate chemotherapy of TB, and to prevent the spread of resistant strains. The aim of this study was to characterize TB patients at higher risk of having MDR-TB, to analyze the drug resistance and epidemiological profile of these patients. Use the clinical samples to assess the commercial test (GenoType® MTBDRplus) and develop and standardize a new test (Detect-MDRTB) for detecting the most frequent mutations associated with resistance to INH and RIF. A significantly higher proportion (75% versus 20%, p = 0.009) of males were found among drug-resistant cases than drug susceptible cases. 43.8% of patients took longer than 30 days to seek medical care and in the MDR group 25% of the cases did not undergo any previous anti-TB treatment. In our samples we found a proportion of 48.3% of MDR-TB. The T family was the most frequent spoligotype family. Compared with the proportion method, the sensitivity and specificity of the MTBDRplus assay were 82% and 94% for RIF-resistance, 60% and 94% for INH resistance. Compared with sequencing, the sensitivity and specificity of the MTBDRplus assay were 92% and 97% for RIF-resistance, 100% and 100% for INHresistance. To detect RIF and INH-resistance, the Detect-TBMDR assay showed a sensitivity and specificity of 79.3% and 77.0%, and 100% and 65%, respectively, compared to proportion method. When compared with sequencing, Detect-TBMDR assay, to detect RIF and INH-resistance, showed a sensitivity and specificity of 81.2% and 94.7% and to 100% and 96.2%, respectively. Discordances still exist between the proportion method and molecular approach, particularly regarding INH-resistance. However, these methods are very important for the management faster and correct patient, helping to choose the best treatment regimen.

Thesis (M.Sc. (Medical Microbiology)) --University Limpopo, 2013 / Genotyping of TB is essential to investigate and confirm transmission of the multi-drug resistant tuberculosis and of great value in optimizing strategies for the determination of strains causing the increased mortality rates of TB outbreaks. Sputum samples (207) were collected from National Health Laboratory Services in Polokwane laboratory for determining mutations and genotypes of the Mycobacterium tuberculosis strains using GenoType®MTBDRplus (Hain LifeScience, Germany) and Real-Time PCR (Roche, South Africa) techniques. Of the 207 samples, 28 (13.5%) exhibited drug resistance. Thirteen of the 28 (46%) MDR-TB strains belonged to the non-Beijing family, with mutations at codons rpoB 516 and rpoB 526 for RIF and katG 315 and inhA 15 for INH resistance. The Non-Beijing strains 75% (21/28) were monoresistant to RIF 14% (3/21) at codons 516, 526, 531 of rpoB gene and INH 19% (4/21) at codon 315 of katG and codon 15 of inhA 5% (1/21). Of the eight Beijing strains, 3(8%) were INH- resistant at codon 315 for katG and codon 15 for inhA and 3(8%) were RIF-resistant with mutations at codons 516 and 526. Two samples were typed as MDR for the Beijing strains with codon 315 for INH and codons 526 and 531 for RIF. The sample with a co-infection for Beijing and non-Beijing was an MDR-TB strain with mutations in rpoB codons 526, 531, katG 315 and inhA 8, 15 and16. The study showed a high rate of drug resistance with the non-Beijing compared to Beijing strains and mutations in specific codons for RIF and INH are variable for the TB families.

Abstract not available