Pesquisa | SciELO

ABSTRACT Background: Brazil has the second largest number of leprosy cases worldwide, and the state of São Paulo has been considered non-endemic since 2006. Methods: We analyzed 16 variable number tandem repeats loci and three single nucleotide polymorphisms loci of Mycobacterium leprae (M. leprae) in 125 clinical isolates from patients in different municipalities in the state. Results: The clustering pattern of M. leprae indicated that the transmission of leprosy persisted in the state and included scenarios of intra-extra-familial transmission in areas with low endemicity. Conclusions: A significantly active circulation of M. leprae was observed. Therefore, surveillance and control measures must be implemented.

https://doi.org/10.1590/0037-8682-0612-2022

Tuberculosis drug resistance profiling based on machine learning: A literature review
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Sharma, Abhinav

; Machado, Edson

; Lima, Karla Valeria Batista

; Suffys, Philip Noel

; Conceição, Emilyn Costa

Brazilian Journal of Infectious Diseases

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2022, Volume 26 Nº 1 elocation 102332

Resumo: En | Texto: En | PDF: En

Abstract Tuberculosis (TB), caused by Mycobacterium tuberculosis (MTB), is one of the top 10 causes of death worldwide. Drug-resistant tuberculosis (DR-TB) poses a major threat to the World Health Organization's “End TB” strategy which has defined its target as the year 2035. In 2019, there were close to 0.5 million cases of DRTB, of which 78% were resistant to multiple TB drugs. The traditional culture-based drug susceptibility test (DST - the current gold standard) often takes multiple weeks and the necessary laboratory facilities are not readily available in low-income countries. Whole genome sequencing (WGS) technology is rapidly becoming an important tool in clinical and research applications including transmission detection or prediction of DR-TB. For the latter, many tools have recently been developed using curated database(s) of known resistance conferring mutations. However, documenting all the mutations and their effect is a time-taking and a continuous process and therefore Machine Learning (ML) techniques can be useful for predicting the presence of DR-TB based on WGS data. This can pave the way to an earlier detection of drug resistance and consequently more efficient treatment when compared to the traditional DST.

https://doi.org/10.1016/j.bjid.2022.102332

Heterogeneous persistence of Mycobacterium leprae in oral and nasal mucosa of multibacillary patients during multidrug therapy
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Neumann, Arthur da Silva

; Fontes, Amanda Nogueira Brum

; Lopes, Márcia Quinhones Pires

; Suffys, Philip Noel

; Moraes, Milton Ozório

; Lara, Flávio Alves

Memórias do Instituto Oswaldo Cruz

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2022, Volume 117 elocation e220058

Resumo: En | Texto: En | PDF: En

BACKGROUND Leprosy is curable by multidrug therapy (MDT) treatment regimen ranging from six to 12 months. The variable levels of tolerance and adherence among patients can, however, result in treatment failure and the emergence of drug-resistant strains. OBJECTIVES Describe the impact of MDT over Mycobacterium leprae viability in patient’s oral and nasal mucosa along treatment. METHODS Mycobacterium leprae viability was monitored by quantitative polymerase chain reaction (qPCR) quantification of 16S rRNA in lateral and contralateral scrapings of oral and nasal mucosa of 10 multibacillary patients along the initial five months of treatment. FINDINGS The results demonstrated high heterogenicity of M. leprae viability among patients and between nasal and oral samples. Of six patients who presented good adherence and tolerance to the treatment, only four displayed absence of M. leprae viability in both samples three months after the first MDT dose, while for the other two, the absence of M. leprae viability in the oral and nasal cavities was only detected five months after the first dose. MAIN CONCLUSIONS We concluded that qPCR of 16S rRNA for the determination of M. leprae viability in nasal and oral scraping samples could represent an interesting approach to monitor treatment efficacy.

https://doi.org/10.1590/0074-02760220058

Molecular epidemiology of Mycobacterium tuberculosis in Brazil before the whole genome sequencing era: a literature review
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Conceição, Emilyn Costa

; Salvato, Richard Steiner

; Gomes, Karen Machado

; Guimarães, Arthur Emil dos Santos

; da Conceição, Marília Lima

; Souza e Guimarães, Ricardo José de Paula

; Sharma, Abhinav

; Furlaneto, Ismari Perini

; Barcellos, Regina Bones

; Bollela, Valdes Roberto

; Anselmo, Lívia Maria Pala

; Sisco, Maria Carolina

; Niero, Cristina Viana

; Ferrazoli, Lucilaine

; Refrégier, Guislaine

; Lourenço, Maria Cristina da Silva

; Gomes, Harrison Magdinier

; de Brito, Artemir Coelho

; Catanho, Marcos

; Duarte, Rafael Silva

; Suffys, Philip Noel

; Lima, Karla Valéria Batista

Memórias do Instituto Oswaldo Cruz

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2021, Volume 116 elocation e200517

Resumo: En | Texto: En | PDF: En

Molecular-typing can help in unraveling epidemiological scenarios and improvement for disease control strategies. A literature review of Mycobacterium tuberculosis transmission in Brazil through genotyping on 56 studies published from 1996-2019 was performed. The clustering rate for mycobacterial interspersed repetitive units - variable tandem repeats (MIRU-VNTR) of 1,613 isolates were: 73%, 33% and 28% based on 12, 15 and 24-loci, respectively; while for RFLP-IS6110 were: 84% among prison population in Rio de Janeiro, 69% among multidrug-resistant isolates in Rio Grande do Sul, and 56.2% in general population in São Paulo. These findings could improve tuberculosis (TB) surveillance and set up a solid basis to build a database of Mycobacterium genomes.

https://doi.org/10.1590/0074-02760200517 744 downloads

Spoligotyping, genotyping, and spatial distribution of Mycobacterium bovis in cattle in the state of Bahia, Brazil
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Santos, Evelin S.V.

; Alzamora Filho, Fernando

; Santos, Bruno R.

; Silva, Alana V.

; Gomes, Harrison M.

; Suffys, Philip N.

; Costa, Joselito N.

Pesquisa Veterinária Brasileira

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2021, Volume 41 elocation e06729

Resumo: Pt En | Texto: Pt En | PDF: Pt | PDF: En

RESUMO: O Mycobacterium bovis é o responsável pela tuberculose bovina e bubalina, doença zoonótica importante e com distribuição global. O conhecimento da distribuição e a identificação precisa dessa enfermidade, incluindo diagnósticos mais avançados como o spoligotyping, permite escolher as melhores estratégias de combate ao avanço da doença. O presente trabalho objetivou investigar a presença de micobactérias, genotipar suas estirpes e avaliar a distribuição espacial dos casos de tuberculose a partir de lesões sugestivas nas carcaças de bovinos e bubalinos inspecionadas em frigoríficos sob regime de inspeção oficial no estado da Bahia. Foram investigados 453.417 animais dentre os quais 31 (0,007%) foram suspeitos de doença e provenientes de 17 municípios. Após o crescimento em meio de cultura, 95% foram categorizados como bacilos álcool-ácido resistentes (BAAR). Todos os isolados foram submetidos à spoligotyping e 95% foram confirmados M. bovis (SB0120, SB0121, SB0852, SB0828, SB0295, SB0881, SB1648, SB6119, SB0140, SB1055). A cepa SB0120 foi a mais prevalente e este perfil vem sendo descrito na literatura com casos de tuberculose humana por M. bovis ressaltando o potencial zoonótico deste perfil. Este estudo também identificou cepas nunca relatadas no estado da Bahia, destacando um padrão distinto de outras partes do Brasil, além da existência de infecções mistas. Permitiu ainda relatar linhagens nunca antes descritas no estado com destaque para um padrão novo no Brasil (SB6119 e SB0852 respectivamente). Um perfil inédito identificado foi identificado e inserido no banco de dados internacional (Mbovis.org), nomeado SB2715.

ABSTRACT: Mycobacterium bovis is responsible for bovine and buffalo tuberculosis, an important zoonotic disease with global distribution. The knowledge of the distribution and the precise identification of this disease, including advanced diagnoses such as spoligotyping, allows choosing the best strategies to fight the disease’s progress. The present work aimed to investigate mycobacteria’s presence, genotype their strains, and evaluate tuberculosis cases’ spatial distribution from suggestive lesions in carcasses of bovine and buffalo inspected in slaughterhouses under an official inspection regime in the state of Bahia, Brazil. The study investigated 453,417 animals. Among these, 31 (0.007%) from 17 municipalities were suspected of tuberculosis. Among the culture medium growth, 95% of these were categorized as alcohol-acid resistant bacilli (BAAR). All isolates were subjected to spoligotyping and 95% were confirmed as M. bovis (SB0120, SB0121, SB0852, SB0828, SB0295, SB0881, SB1648, SB6119, SB0140, SB1055). The strain SB0120 was the most prevalent, and this profile has been described in cases of human tuberculosis by M. bovis, highlighting the zoonotic potential of this profile. This study also identified strains never reported in Bahia, highlighting a distinctive pattern from other parts of Brazil, besides mixed infections. Besides, to identify strains never before described in the state, highlighting a distinctive pattern in Brazil (SB6119 and SB0852, respectively). An unpublished profile was identified and inserted in the international database (Mbovis.org), named SB2715.

https://doi.org/10.1590/1678-5150-pvb-6729 197 downloads

Evaluation of drug susceptibility profile of Mycobacterium tuberculosis Lineage 1 from Brazil based on whole genome sequencing and phenotypic methods
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Guimarães, Arthur Emil dos Santos

; Sharma, Abhinav

; Furlaneto, Ismari Perini

; Rutaihwa, Liliana

; Cardoso, Jedson Ferreira

; da Conceição, Marília Lima

; Spinassé, Lizânia Borges

; Machado, Edson

; Lopes, Maria Luiza

; Duarte, Rafael Silva

; Gagneux, Sebastien

; Suffys, Philip Noel

; Lima, Karla Valéria Batista

; Conceição, Emilyn Costa

Memórias do Instituto Oswaldo Cruz

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2020, Volume 115 elocation e200520

Resumo: En | Texto: En | PDF: En

BACKGROUND The evaluation of procedures for drug susceptibility prediction of Mycobacterium tuberculosis based on genomic data against the conventional reference method test based on culture is realistic considering the scenario of growing number of tools proposals based on whole-genome sequences (WGS). OBJECTIVES This study aimed to evaluate drug susceptibility testing (DST) outcome based on WGS tools and the phenotypic methods performed on isolates of M. tuberculosis Lineage 1 from the state of Pará, Brazil, generally associated with low levels of drug resistance. METHODOLOGY Culture based DST was performed using the Proportion Method in Löwenstein-Jensen medium on 71 isolates that had been submitted to WGS. We analysed the seven main genome sequence-based tools for resistance and lineage prediction applied to M. tuberculosis and for comparison evaluation we have used the Kappa concordance test. FINDINGS When comparing the WGS-based tools against the DST, we observed the highest level of agreement using TB-profiler. Among the tools, TB-profiler, KvarQ and Mykrobe were those which identified the largest number of TB-MDR cases. Comparing the four most sensitive tools regarding resistance prediction, agreement was observed for 43 genomes. MAIN CONCLUSIONS Drug resistance profiling using next-generation sequencing offers rapid assessment of resistance-associated mutations, therefore facilitating rapid access to effective treatment.

https://doi.org/10.1590/0074-02760200520 427 downloads

Newly sequenced genomes of four Bacillus Calmette Guerin vaccines
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Sisco, Maria Carolina

; Silva, Marlei Gomés

; Lopez, Beatriz

; Arguelles, Claudia

; Mendonça-Lima, Leila

; de Waard, Jacobus H

; Duarte, Rafael Silva

; Suffys, Philip Noel

Memórias do Instituto Oswaldo Cruz

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2020, Volume 115 elocation e190401

Resumo: En | Texto: En | PDF: En

Bacillus Calmette Guerin (BCG) vaccines comprise a family of related strains. Whole genome sequencing has allowed the better characterisation of the differences between many of the BCG vaccines. As sequencing technologies improve, updating of publicly available sequence data becomes common practice. We hereby announce the draft genome of four commonly used BCG vaccines in Brazil, Argentina and Venezuela.

https://doi.org/10.1590/0074-02760190401 669 downloads

Antiretroviral therapy-induced paradoxical worsening of previously healed Mycobacterium haemophilum cutaneous lesions in advanced HIV infection
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Eyer-Silva, Walter de Araujo

; Almeida, Marina Rodrigues de

; Martins, Carlos José

; Basílio-de-Oliveira, Rodrigo Panno

; Araujo, Luciana Ferreira de

; Basílio-de-Oliveira, Carlos Alberto

; Azevedo, Marcelo Costa Velho Mendes de

; Pinto, Jorge Francisco da Cunha

; Vasconcellos, Sidra Ezidio Gonçalves

; Rodrigues-dos-Santos, Ícaro

; MagdinierGomes, Harrison

; Suffys, Philip Noel

Revista do Instituto de Medicina Tropical de São Paulo

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2019, Volume 61 elocation e71

Resumo: En | Texto: En | PDF: En

ABSTRACT Mycobacterium haemophilum is a nontuberculous mycobacterium that causes localized or disseminated disease, mainly in immunocompromised hosts. We report the case of a 35-year-old HIV-infected woman who presented with several enlarging cutaneous lesions over the arms and legs. Histopathological examination revealed the diagnosis of a cutaneous mycobacterial disease. Mycobacterial analyses unveiled M. haemophilum infection. Six months after completion of a successful antimycobacterial treatment, she developed an immune reconstitution inflammatory syndrome (IRIS). This paradoxical relapse presented as tenderness, redness and swelling at the precise sites of the healed lesions and took place in the setting of significant recovery of the CD4 cell count (from 05 to 318 cells/mm 3 ). Microbiological analyses of these worsening lesions were negative, and they spontaneously remitted without the initiation of a novel antimycobacterial treatment cycle. M. haemophilum infection should always be considered as a cause of skin lesions in immunocompromised subjects. Physicians should be aware of the possibility of IRIS as a complication of successful antiretroviral therapy in HIV-infected patients with M. haemophilum infection.

https://doi.org/10.1590/s1678-9946201961071 412 downloads

Comparative study of RFLP-IS6110 and MIRU-VNTR from Mycobacterium tuberculosis isolated in the state of Minas Gerais, Brazil
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Augusto, Cláudio José

; Carvalho, Wânia da Silva

; Almeida, Isabela Neves de

; Figueiredo, Lida Jouca de Assis

; Dantas, Nayanne Gama Teixeira

; Suffys, Philip Noel

; Miranda, Silvana Spíndola de

Brazilian Journal of Microbiology

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Set 2018, Volume 49 Nº 3 Páginas 641 - 646

Resumo: En | Texto: En | PDF: En

Abstract DNA genotyping of Mycobacterium tuberculosis has been widely applied in the understanding of disease transmission in many countries. The purpose of this study was to genotype the strains of M. tuberculosis isolated in patients with new tuberculosis (TB) cases in Minas Gerais, as well as to compare the similarity, discriminatory power, and agreement of the clusters between the IS6110 Restriction Fragment Length Polymorfism (RFLP) and 12 loci Variable Number Tandem Repeat - Mycobacterial Interspersed Repetitive Units (MIRU-VNTR) techniques. It was observed that 32% (66/204) of the isolated strains in the RFLP-IS6110 and 50.9% (104/204) of the isolated strains in the MIRU-VNTR presented a similarity of equal to or above 85%. The RFLP-IS6110 and MIRU-VNTR proved to contain a high discriminatory power. The similarity index resulting from the RFLP showed no recent transmission. Good agreement was observed between the techniques when clusters were detected; however, the best epidemiological relationship was found when using the RFLP-IS6110.

https://doi.org/10.1016/j.bjm.2017.04.015 806 downloads

10.

Whole genome sequence of Mycobacterium kansasii isolates of the genotype 1 from Brazilian patients with pulmonary disease demonstrates considerable heterogeneity
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Machado, Edson

; Vasconcellos, Sidra Ezidio Gonçalves

; Cerdeira, Camillo

; Gomes, Lia Lima

; Junqueira, Ricardo

; Carvalho, Luciana Distasio de

; Ramos, Jesus Pais

; Redner, Paulo

; Campos, Carlos Eduardo Dias

; Caldas, Paulo Cesar de Souza

; Gomes, Ana Paula Chaves Sobral

; Goldenberg, Telma

; Montes, Fatima Fandinho

; Mello, Fernanda Carvalho de Queiroz

; Mussi, Vinicius de Oliveira

; Lasunskaia, Elena

; Soolingen, Dick van

; Miranda, Antonio Basílio de

; Rigouts, Leen

; Jong, Bouke C de

; Meehan, Conor J

; Catanho, Marcos

; Suffys, Philip N

Memórias do Instituto Oswaldo Cruz

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2018, Volume 113 Nº 9 elocation e180085

Resumo: En | Texto: En | PDF: En

Mycobacterium kansasii is an opportunistic pathogen and one of the most commonly encountered species in individuals with lung disease. We here report the complete genome sequence of 12 clinical isolates of M. kansasii from patients with pulmonary disease in Brazil.

https://doi.org/10.1590/0074-02760180085 1130 downloads

11.

Risk factors for human Mycobacterium bovis infections in an urban area of Brazil
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Silva, Marcio Roberto

; Rocha, Adalgiza da Silva

; Araújo, Flábio Ribeiro

; Fonseca-Júnior, Antônio Augusto

; Alencar, Andrea Padilha de

; Suffys, Philip Noel

; Costa, Ronaldo Rodrigues da

; Moreira, Maria Aparecida Scatamburlo

; Guimarães, Mark Drew Crosland

Memórias do Instituto Oswaldo Cruz

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2018, Volume 113 Nº 8 elocation e170445

Resumo: En | Texto: En | PDF: En

BACKGROUND The World Health Organization (WHO) has classified human zoonotic tuberculosis (TB) due to Mycobacterium bovis as a neglected issue in the developing world. In a recent cross-sectional study in Brazil, three of 189 TB patients presented with a coinfection of M. bovis and M. tuberculosis and were selected as cases for this study. OBJECTIVE The aim was to evaluate risk factors (RF) for zoonotic TB in an urban area of Brazil in order to guide preventive programmes. METHODS A matched case-control study was carried out nested within a cross-sectional study. For each of the three cases, 14 age- and sex-matched controls (TB due to M. tuberculosis) were selected. FINDINGS Zoonotic potential exposures (ZE) and extrapulmonary TB (EPTB) were independently associated with zoonotic TB in multivariate analyses. CONCLUSIONS ZE by occupation and consumption of raw milk and derivative products that place individuals in direct and indirect contact with animals and their excretions/secretions increase the risk for zoonotic TB in Brazil, especially among those with EPTB. Therefore, measures such as efficient control of bovine TB, distribution of pasteurised milk and its derivative products, and the diagnosis and monitoring of zoonotic TB in humans are essential steps, especially in developing countries where bovine TB is enzootic, and further studies are necessary.

https://doi.org/10.1590/0074-02760170445 2022 downloads

12.

Tuberculosis associated factors caused by Mycobacterium tuberculosis of the RDRio genotype
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Moraes, Eloise Brasil

; Slompo, Letícia

; Finardi, Amanda Juliane

; Silveira, Heloisa Paro Pedro da

; Ruiz, Luciana

; Gomes, Harrison Magdinier

; Richini, Virginia Bodelão

; Suffys, Philip

; Fortaleza, Carlos Magno Castelo Branco

; Cavalcanti, Ricardo

; Baptista, Ida Maria Foschiani Dias

Memórias do Instituto Oswaldo Cruz

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Mar 2017, Volume 112 Nº 3 Páginas 182 - 187

Resumo: En | Texto: En | PDF: En

BACKGROUND Tuberculosis (TB) continues to be a disease that affects many countries around the world, including Brazil. Recently, a subtype of Latin American-Mediterranean family strain was identified and characterised by RDRio. The strain has been associated with different characteristics of the disease. OBJECTIVES In the present study we investigated the association of epidemiological, clinical, radiological and bacteriological variables with pulmonary tuberculosis caused by RDRioMycobacterium tuberculosis strain in large regions of São Paulo. METHODS We conducted a cross-sectional study in 530 patients with pulmonary tuberculosis, diagnosed using sputum culture, from two regions of the São Paulo state in Brazil. The samples were brought to São Paulo reference laboratories for epidemiological, clinical, radiological and bacteriological analyses, and the data were obtained from a TB notification system. RDRio genotyping and Spoligotyping of the samples were performed. For the analysis of the categorical variables we used the chi-square test or the Fisher’s exact test, and for the continuous variables, the Mann-Whitney test. In addition, a logistic regression was used for multivariate analysis. Differences with p < 0.05 were considered significant. FINDINGS The RDRio deletion was identified in 152 (28.7%) samples. In the univariate analysis, both the age groups above 25 years and alcohol consumption were associated with the RDRio deletion. The multivariate analysis confirmed the association of the RDRio deletion with the age groups: 25-35 years old [OR: 2.28 (1.02-5.07; p = 0.04)] and 36-60 years old (OR: 2.36 (1.11-5.05); p = 0.03], and also with alcohol consumption [OR: 1.63 (1.05-2.54); p = 0,03]. MAIN CONCLUSIONS In this study, we identified new factors associated with the M. tuberculosis of the RDRio deletion strains infection.

https://doi.org/10.1590/0074-02760160347 1157 downloads

13.

Clinical and epidemiological profiles of individuals with drug-resistant tuberculosis
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Pedro, Heloisa da Silveira Paro

; Nardi, Susilene Maria Tonelli

; Pereira, Maria Izabel Ferreira

; Oliveira, Rosângela Siqueira

; Suffys, Philip Noel

; Gomes, Harrison Magdinier

; Finardi, Amanda Juliane

; Moraes, Eloise Brasil de

; Baptista, Ida Maria Foschiani Dias

; Machado, Ricardo Luiz Dantas

; Castiglioni, Lilian

Memórias do Instituto Oswaldo Cruz

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Abr 2015, Volume 110 Nº 2 Páginas 235 - 248

Resumo: En | Texto: En | PDF: En

Drug-resistant tuberculosis (TB) is a growing global threat. Approximately 450,000 people developed multidrug-resistant TB worldwide in 2012 and an estimated 170,000 people died from the disease. This paper describes the sociodemographic, clinical-epidemiological and bacteriological aspects of TB and correlates these features with the distribution of anti-TB drug resistance. Mycobacterium tuberculosis (MT) cultures and drug susceptibility testing were performed according to the BACTEC MGIT 960 method. The results demonstrated that MT strains from individuals who received treatment for TB and people who were infected with human immunodeficiency virus were more resistant to TB drugs compared to other individuals (p < 0.05). Approximately half of the individuals received supervised treatment, but most drug-resistant cases were positive for pulmonary TB and exhibited positive acid-fast bacilli smears, which are complicating factors for TB control programs. Primary healthcare is the ideal level for early disease detection, but tertiary healthcare is the most common entry point for patients into the system. These factors require special attention from healthcare managers and professionals to effectively control and monitor the spread of TB drug-resistant cases.

https://doi.org/10.1590/0074-02760140316 2214 downloads

14.

In house reverse membrane hybridisation assay versus GenoType MTBDRplus and their performance to detect mutations in the genes rpoB, katG and inhA
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Ferreira Junior, Sergio Luiz Montego

; Costa, Elis Regina Dalla

; Santos, Paula Gonçalves dos

; Gomes, Harrison Magdinier

; Silva, Marcia Susana Nunes

; Esteves, Leonardo Souza

; Oliveira, Martha Maria

; Maschmann, Raquel de Abreu

; Kritski, Afrânio Lineu

; Suffys, Philip Noel

; Rossetti, Maria Lucia Rosa

Memórias do Instituto Oswaldo Cruz

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Jun 2014, Volume 109 Nº 3 Páginas 307 - 314

Resumo: En | Texto: En | PDF: En

Drug-resistant tuberculosis (TB) threatens global TB control and is a major public health concern in several countries. We therefore developed a multiplex assay (LINE-TB/MDR) that is able to identify the most frequent mutations related to rifampicin (RMP) and isoniazid (INH) resistance. The assay is based on multiplex polymerase chain reaction, membrane hybridisation and colorimetric detection targeting of rpoB and katG genes, as well as the inhA promoter, which are all known to carry specific mutations associated with multidrug-resistant TB (MDR-TB). The assay was validated on a reference panel of 108 M. tuberculosis isolates that were characterised by the proportion method and by DNA sequencing of the targets. When comparing the performance of LINE-TB/MDR with DNA sequencing, the sensitivity, specificity and agreement were 100%, 100% and 100%, respectively, for RMP and 77.6%, 90.6% and 88.9%, respectively, for INH. Using drug sensibility testing as a reference standard, the performance of LINE-TB/MDR regarding sensitivity, specificity and agreement was 100%, 100% and 100% (95%), respectively, for RMP and 77%, 100% and 88.7% (82.2-95.1), respectively, for INH. LINE-TB/MDR was compared with GenoType MTBDRplus for 65 isolates, resulting in an agreement of 93.6% (86.7-97.5) for RIF and 87.4% (84.3-96.2) for INH. LINE-TB/MDR warrants further clinical validation and may be an affordable alternative for MDR-TB diagnosis.

https://doi.org/10.1590/0074-0276130469 1524 downloads

15.

Direct detection of Mycobacterium tuberculosis complex in bovine and bubaline tissues through nested-PCR
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Araújo, Cristina P.

; Osório, Ana Luiza A.R.

; Jorge, Klaudia S.G.

; Ramos, Carlos A.N.

; Souza Filho, Antonio F.

; Vidal, Carlos E.S.

; Vargas, Agueda P.C.

; Roxo, Eliana

; Rocha, Adalgiza S.

; Suffys, Philip N.

; Fonseca Júnior, Antônio A.

; Silva, Marcio R.

; Barbosa Neto, José D.

; Cerqueira, Valíria D.

; Araújo, Flábio R.

Brazilian Journal of Microbiology

Métricas do periódico
- Sobre o periódico
- SciELO Analytics

Jun 2014, Volume 45 Nº 2 Páginas 633 - 640

Resumo: En | Texto: En | PDF: En

Post-mortem bacterial culture and specific biochemical tests are currently performed to characterize the etiologic agent of bovine tuberculosis. Cultures take up to 90 days to develop. A diagnosis by molecular tests such as PCR can provide fast and reliable results while significantly decreasing the time of confirmation. In the present study, a nested-PCR system, targeting rv2807, with conventional PCR followed by real-time PCR, was developed to detect Mycobacterium tuberculosis complex (MTC) organisms directly from bovine and bubaline tissue homogenates. The sensitivity and specificity of the reactions were assessed with DNA samples extracted from tuberculous and non-tuberculous mycobacteria, as well as other Actinomycetales species and DNA samples extracted directly from bovine and bubaline tissue homogenates. Regarding the analytical sensitivity, DNA of the M. bovis AN5 strain was detected up to 1.5 pg by nested-PCR, whereas DNA of M. tuberculosis H37Rv strain was detected up to 6.1 pg. The nested-PCR system showed 100% analytical specificity for MTC when tested with DNA of reference strains of non-tuberculous mycobacteria and closely-related Actinomycetales. A clinical sensitivity level of 76.7% was detected with tissues samples positive for MTC by means of the culture and conventional PCR. A clinical specificity of 100% was detected with DNA from tissue samples of cattle with negative results in the comparative intradermal tuberculin test. These cattle exhibited no visible lesions and were negative in the culture for MTC. The use of the nested-PCR assay to detect M. tuberculosis complex in tissue homogenates provided a rapid diagnosis of bovine and bubaline tuberculosis.

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