A comparison of mycolic acid analysis for nontuberculous mycobacteria identification by thin-layer chromatography and molecular methods

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dc.contributor.authorFujimura Leite, Clarisse Queico [UNESP]
dc.contributor.authorDa Silva Rocha, Adalgiza
dc.contributor.authorDe Andrade Leite, Sergio Roberto [UNESP]
dc.contributor.authorCarvalho Ferreira, Rosa Maria
dc.contributor.authorSuffys, Philip N.
dc.contributor.authorFonseca, Lella de Souza
dc.contributor.authorFéres Saad, Maria Helena
dc.contributor.institutionUniversidade Estadual Paulista (Unesp)
dc.contributor.institutionOswaldo Cruz Institute
dc.contributor.institutionUniversidade Federal do Rio de Janeiro (UFRJ)
dc.contributor.institutionFIOCRUZ
dc.date.accessioned2014-05-27T11:21:37Z
dc.date.available2014-05-27T11:21:37Z
dc.date.issued2005-09-21
dc.description.abstractThe development of fast, inexpensive, and reliable tests to identify nontuberculous mycobacteria (NTM) is needed. Studies have indicated that the conventional identification procedures, including biochemical assays, are imprecise. This study evaluated a proposed alternative identification method in which 83 NTM isolates, previously identified by conventional biochemical testing and in-house M. avium IS1245-PCR amplification, were submitted to the following tests: thin-layer chromatography (TLC) of mycolic acids and PCR-restriction enzyme analysis of hsp65 (PRA). High-performance liquid chromatography (HPLC) analysis of mycolic acids and Southern blot analysis for M. avium IS1245 were performed on the strains that evidenced discrepancies on either of the above tests. Sixty-eight out of 83 (82%) isolates were concordantly identified by the presence of IS1245 and PRA and by TLC mycolic acid analysis. Discrepant results were found between the phenotypic and molecular tests in 12/83 (14.4%) isolates. Most of these strains were isolated from non-sterile body sites and were most probably colonizing in the host tissue. While TLC patterns suggested the presence of polymycobacterial infection in 3/83 (3.6%) cultures, this was the case in only one HPLC-tested culture and in none of those tested by PRA. The results of this study indicated that, as a phenotypic identification procedure, TLC mycolic acid determination could be considered a relatively simple and cost-effective method for routine screening of NTM isolates in mycobacteriology laboratory practice with a potential for use in developing countries. Further positive evidence was that this method demonstrated general agreement on MAC and M. simiae identification, including in the mixed cultures that predominated in the isolates of the disseminated infections in the AIDS patients under study. In view of the fact that the same treatment regimen is recommended for infections caused by these two species, TLC mycolic acid analysis may be a useful identification tool wherever molecular methods are unaffordable.en
dc.description.affiliationDepartment of Pharmacy Institute of Chemistry UNESP, Araraquara, São Paulo
dc.description.affiliationInstitute of Chemistry UNESP, Araraquara, São Paulo
dc.description.affiliationDepartment of Mycobacterioses Evandro Chagas Hospital Oswaldo Cruz Institute, Fiocruz, Rio de Janeiro, R.J.
dc.description.affiliationEvandro Chagas Hospital Oswaldo Cruz Institute, Fiocruz, Rio de Janeiro, R.J.
dc.description.affiliationMycobacteriology Laboratory Microbiology Institute UFRJ, Rio de Janeiro, R.J.
dc.description.affiliationMicrobiology Institute UFRJ, Rio de Janeiro, R.J.
dc.description.affiliationDepartment of Mycobacterioses Instituto Oswaldo Cruz FIOCRUZ, Av. Brasil 4365, 21045-900 Rio de Janeiro, RJ
dc.description.affiliationUnespDepartment of Pharmacy Institute of Chemistry UNESP, Araraquara, São Paulo
dc.description.affiliationUnespInstitute of Chemistry UNESP, Araraquara, São Paulo
dc.format.extent571-578
dc.identifierhttp://dx.doi.org/10.1111/j.1348-0421.2005.tb03642.x
dc.identifier.citationMicrobiology and Immunology, v. 49, n. 7, p. 571-578, 2005.
dc.identifier.doi10.1111/j.1348-0421.2005.tb03642.x
dc.identifier.issn0385-5600
dc.identifier.scopus2-s2.0-24744437725
dc.identifier.urihttp://hdl.handle.net/11449/68414
dc.identifier.wosWOS:000230410300001
dc.language.isoeng
dc.relation.ispartofMicrobiology and Immunology
dc.relation.ispartofjcr1.335
dc.relation.ispartofsjr0,764
dc.rights.accessRightsAcesso restrito
dc.sourceScopus
dc.subjectIS1245
dc.subjectMycolic acids
dc.subjectNontuberculous mycobacteria
dc.subjectPRA
dc.subjectTLC
dc.subjectheat shock protein 65
dc.subjectmycolic acid
dc.subjectacquired immune deficiency syndrome
dc.subjectatypical Mycobacterium
dc.subjectbacterial colonization
dc.subjectbacterial strain
dc.subjectbacterium culture
dc.subjectbacterium detection
dc.subjectbacterium identification
dc.subjectbacterium isolate
dc.subjectbacterium isolation
dc.subjectcontrolled study
dc.subjectcost effectiveness analysis
dc.subjectenzyme assay
dc.subjecthigh performance liquid chromatography
dc.subjectintermethod comparison
dc.subjectmixed cell culture
dc.subjectMycobacterium avium
dc.subjectnonhuman
dc.subjectphenotype
dc.subjectpolymerase chain reaction
dc.subjectrestriction mapping
dc.subjectscreening test
dc.subjectSouthern blotting
dc.subjectsuperinfection
dc.subjectthin layer chromatography
dc.subjectBacterial Typing Techniques
dc.subjectChromatography, Thin Layer
dc.subjectDNA, Bacterial
dc.subjectMycobacterium
dc.subjectMycolic Acids
dc.subjectatypical mycobacterium
dc.subjectBacteria (microorganisms)
dc.subjectCorynebacterineae
dc.titleA comparison of mycolic acid analysis for nontuberculous mycobacteria identification by thin-layer chromatography and molecular methodsen
dc.typeArtigo
dcterms.licensehttp://olabout.wiley.com/WileyCDA/Section/id-406071.html

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