Identification of Mycobacteria by Thin Layer Chromatographic Analysis of Mycolic Acids and Conventional Biochemical Method: Four Years of Experience

Leite, Clarice Queico Fujimura; Souza, Clóvis Wesley Oliveira de; Leite, Sérgio Roberto de Andrade

doi:10.1590/s0074-02761998000600019

Cited by 32 publications

(35 citation statements)

References 12 publications

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“…The cells from eugonic and dysgonic colonies that suggest the growth of mycobacteria were examined microscopically after Ziehl-Neelsen staining for acid-fast bacilli. Eugonic mycobacterial isolates were identified by conventional methods (rate of growth, colonial morphology and pigmentation, and biochemical properties) (Brasil 1994), and analysis of mycolic acid profiles using thinlayer chromatography (TLC) (Leite et al 1998). Unusually slowly growing dysgonic cultures, that required 50-60 days for the appearance of colonies, limited to one or two in each Stonebrink medium tube, were submitted to PCR analysis.…”

Section: Methodsmentioning

confidence: 99%

“…Mycolic acids analysis -Extraction and identification of mycolic acids from reference and isolated strains were performed according to Leite et al (1998). The identification of mycolic acids, as methyl esters, was done by onedimensional TLC, with silica gel as the stationary phase and two different elution systems: diethyl ether/petroleum ether 12:88 v/v (three developments), and dichloromethane (single development).…”

Section: Methodsmentioning

confidence: 99%

See 1 more Smart Citation

Isolation and identification of mycobacteria from livestock specimens and milk obtained in Brazil

Leite

Anno

Leite³

et al. 2003

Mem. Inst. Oswaldo Cruz

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Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Isolation and identification of mycobacteria from livestock specimens and milk obtained in Brazil

Leite

Anno

Leite³

et al. 2003

Mem. Inst. Oswaldo Cruz

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“…The incidence detected during avian necropsy has ranged from 0.5% to 14% (21,25,31,32). Mycobacterial infection prevalence in wild birds has not been determined, although usually at least 1 per cent of birds examined postmortem are affected.…”

Section: Epidemiologymentioning

confidence: 99%

“…However, this delays its clinical usefulness in many situations because the culture is not available in less than two weeks and some species, such as M. genavense, are difficult to isolate by standard culture techniques (24,32,51,54). The enzymatic linked immunoassay (ELISA) has been used for detecting antimycobacterial antibodies in the serum of certain species having clinical significance (9,17,42).…”

Section: Diagnosismentioning

confidence: 99%

Mycobacteriosis in Wild Birds: the Potential risk of Disseminating a Little-known Infectious Disease

Soler¹,

Brieva²,

Ribón³

2009

Rev. salud pública

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Avian mycobacteriosis is important for animal and human health; wild birds play an important role in mycobacterial species' ecology and movement. This review was aimed at reporting the role of birds in the spread of avian mycobacteriosis in human and animal populations at risk and thus a systematic review was made of PubMed, Science Direct, Scielo and Scirus databases. Mycobacteria are classified into the Mycobacterium tuberculosis complex and non-tuberculous mycobacteria; the Mycobacterium avium complex represents the most important part of the latter because it is primarily responsible for mycobacterial infection in wild birds and is a potential pathogen for mammals, especially for immunocompromised patients. The clinical signs in birds are variable as it is a chronic and debilitating disease, involving emaciated carcasses, white nodules in different organs and microscopically it presents granulomatosous multifocal inflammation. Diagnosis begins by suspicion based on clinical signs and finishes with microbiological confirmation. New diagnostic techniques include testing with DNA-RNA probes. No effective treatment is currently available and chemoprophylaxis on suspicion of infection is not recommended at the start; these factors increase the potential risk of mycobacteriosis becoming one of the most frequently documented zoonotic diseases which is difficult to treat in birds and humans. Recent concern regarding mycobacterial infection lies in the increased frequency of these opportunistic infections occurring in immunocompromised individuals and these infections' potential impact on bird conservation, this being increased by greater contact between humans and wild and captive birds.

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“…De 1873 até o presente, técnicas que vão desde a taxonomia numérica (Tsukamura, 1966), métodos radiométricos associados à hibridização de sondas de DNA (Ellner et al, 1988), perfis de ácidos micólicos (Leite et al, 1998), seqüênciamento de DNA (Springer et al, 1996;Pai et al, 1997), e testes baseados na PCR, permitiram a identificação de 152 espécies e subespécies no gênero Mycobacterium. As primeiras espécies descobertas foram aquelas que mais afetaram e ainda afetam a saúde do homem: o bacilo de Hansen (1873), denominado de Mycobacterium leprae, e o bacilo de Koch (1882) designado como M. tuberculosis.…”

Section: Introductionunclassified

Pesquisa de Micobactérias Ambientais em água de torneira, luvas e soluções utilizadas em procedimentos cirúrgicos no Hospital Universitário Getúlio Vargas - Manaus/AM

et al. 2009

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RESUMOInvestigou-se por métodos bacteriológicos (cultivo) e moleculares (PCR -Restriction Enzyme Analysis, PRA), a presença de micobactérias ambientais em águas de torneira, soluções e luvas cirúrgicas, utilizadas nas etapas dos procedimentos cirúrgicos executados no centro cirúrgico do Hospital Universitário Getulio Vargas (HUGV), na cidade de Manaus-Amazonas/Brasil. Foram colhidas e analisadas 105 amostras sendo: 24 de águas (colhidas das 2 torneiras existentes no centro cirúrgico), 8 de solução de Povidine e 7 de solução de Clorhexidina, que servem para a higienização das mãos dos cirurgiões; 39 de luvas cirúrgicas (superfícies internas e externas); e 27 de soluções que foram efetivamente utilizadas durante o ato cirúrgico. Por método bacteriológico obteve-se 41 isolados micobacterianos apenas de águas das torneiras. Pelo PRA obteve-se a detecção de DNA micobacteriano somente na amostra de água que forneceu acima de 100 colônias de micobactérias por tubo semeado. Os isolados foram identificados como sendo Mycobacterium celatum perfil 2, M. gordonae perfil 3, M. gordonae perfil 6, M. intracellulare perfil 1, M. lentiflavum perfil 3 e M. mucogenicum perfil 1. O encontro de M. mucogenicum, espécie já incriminada em surtos pós-cirúrgicos, indica que devem ser efetuados procedimentos de limpeza e monitoramento em todos os pontos de distribuição de águas, visando à prevenção de surtos de micobacterioses nosocomiais induzidos pelo uso das águas nas diferentes atividades de manuseio ou higienização dos pacientes submetidos a procedimentos invasivos. PALAVRAS-CHAVE:Micobactérias Ambientais, MNT, água, soluções anti-sépticas, luvas, procedimentos cirúrgicos. Investigation of Environmental Mycobacteria in tap water, surgical gloves and antiseptic solutions used in surgical procedures at the Getúlio Vargas University Hospital, Manaus-AM/BrazilABSTRACT Using bacteriological (culture) and molecular (PCR -Restriction Enzyme Analysis, PRA) methods, we investigated the presence of environmental mycobacteria in tap water, antiseptic solutions and surgical gloves, used in carrying out surgical procedures at the Getúlio Vargas University Hospital Surgical Center, in Manaus -Amazonas/Brazil. Samples (105) were collected and analyzed from: tap water (24 -collected from 3 taps in the surgical center); povidone-iodine solution, PVP-I, (8); Chlorhexidine solution (7), that were used to hygienize surgeons' hands; surgical gloves (39); and solutions that were effectively used during the surgical procedure (27). Using bacteriological method 41 mycobacteria were isolated, all from samples of tap water. Using PRA, mycobacterial DNA was detected only in the sample from water that provided over 100 colonies per inoculated culture tubes. The isolated were identified as Mycobacterium celatum pattern II, M. gordonae pattern III, M. gordonae pattern VI, M. intracellulare pattern I, M. lentiflavum pattern III and M. mucogenicum pattern I. The isolating of M. mucogenicum, a species that had already been incriminated for causing post-surgical outb...

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Identification of Mycobacteria by Thin Layer Chromatographic Analysis of Mycolic Acids and Conventional Biochemical Method: Four Years of Experience

Abstract: Mycolic acids analysis by thin-layer chromatography (TLC)

Cited by 32 publications

References 12 publications

Isolation and identification of mycobacteria from livestock specimens and milk obtained in Brazil

Isolation and identification of mycobacteria from livestock specimens and milk obtained in Brazil

Mycobacteriosis in Wild Birds: the Potential risk of Disseminating a Little-known Infectious Disease

Pesquisa de Micobactérias Ambientais em água de torneira, luvas e soluções utilizadas em procedimentos cirúrgicos no Hospital Universitário Getúlio Vargas - Manaus/AM

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