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Evaluation of amplified rDNA restriction analysis (ARDRA) for the identification of cultured mycobacteria in a diagnostic laboratory

BACKGROUND: The development of DNA amplification for the direct detection of M. tuberculosis from clinical samples has been a major goal of clinical microbiology during the last ten years. However, the limited sensitivity of most DNA amplification techniques restricts their use to smear positive sam...

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Autores principales: De Baere, Thierry, de Mendonça, Ricardo, Claeys, Geert, Verschraegen, Gerda, Mijs, Wouter, Verhelst, Rita, Rottiers, Sylvianne, Van Simaey, Leen, De Ganck, Catharine, Vaneechoutte, Mario
Formato: Texto
Lenguaje:English
Publicado: BioMed Central 2002
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC101405/
https://www.ncbi.nlm.nih.gov/pubmed/11945178
http://dx.doi.org/10.1186/1471-2180-2-4
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author De Baere, Thierry
de Mendonça, Ricardo
Claeys, Geert
Verschraegen, Gerda
Mijs, Wouter
Verhelst, Rita
Rottiers, Sylvianne
Van Simaey, Leen
De Ganck, Catharine
Vaneechoutte, Mario
author_facet De Baere, Thierry
de Mendonça, Ricardo
Claeys, Geert
Verschraegen, Gerda
Mijs, Wouter
Verhelst, Rita
Rottiers, Sylvianne
Van Simaey, Leen
De Ganck, Catharine
Vaneechoutte, Mario
author_sort De Baere, Thierry
collection PubMed
description BACKGROUND: The development of DNA amplification for the direct detection of M. tuberculosis from clinical samples has been a major goal of clinical microbiology during the last ten years. However, the limited sensitivity of most DNA amplification techniques restricts their use to smear positive samples. On the other hand, the development of automated liquid culture has increased the speed and sensitivity of cultivation of mycobacteria. We have opted to combine automated culture with rapid genotypic identification (ARDRA: amplified rDNA restriction analysis) for the detection resp. identification of all mycobacterial species at once, instead of attempting direct PCR based detection from clinical samples of M. tuberculosis only. RESULTS: During 1998–2000 a total of approx. 3500 clinical samples was screened for the presence of M. tuberculosis. Of the 151 culture positive samples, 61 were M. tuberculosis culture positive. Of the 30 smear positive samples, 26 were M. tuberculosis positive. All but three of these 151 mycobacterial isolates could be identified with ARDRA within on average 36 hours. The three isolates that could not be identified belonged to rare species not yet included in our ARDRA fingerprint library or were isolates with an aberrant pattern. CONCLUSIONS: In our hands, automated culture in combination with ARDRA provides with accurate, practically applicable, wide range identification of mycobacterial species. The existing identification library covers most species, and can be easily updated when new species are studied or described. The drawback is that ARDRA is culture-dependent, since automated culture of M. tuberculosis takes on average 16.7 days (range 6 to 29 days). However, culture is needed after all to assess the antibiotic susceptibility of the strains.
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spelling pubmed-1014052002-04-12 Evaluation of amplified rDNA restriction analysis (ARDRA) for the identification of cultured mycobacteria in a diagnostic laboratory De Baere, Thierry de Mendonça, Ricardo Claeys, Geert Verschraegen, Gerda Mijs, Wouter Verhelst, Rita Rottiers, Sylvianne Van Simaey, Leen De Ganck, Catharine Vaneechoutte, Mario BMC Microbiol Methodology Article BACKGROUND: The development of DNA amplification for the direct detection of M. tuberculosis from clinical samples has been a major goal of clinical microbiology during the last ten years. However, the limited sensitivity of most DNA amplification techniques restricts their use to smear positive samples. On the other hand, the development of automated liquid culture has increased the speed and sensitivity of cultivation of mycobacteria. We have opted to combine automated culture with rapid genotypic identification (ARDRA: amplified rDNA restriction analysis) for the detection resp. identification of all mycobacterial species at once, instead of attempting direct PCR based detection from clinical samples of M. tuberculosis only. RESULTS: During 1998–2000 a total of approx. 3500 clinical samples was screened for the presence of M. tuberculosis. Of the 151 culture positive samples, 61 were M. tuberculosis culture positive. Of the 30 smear positive samples, 26 were M. tuberculosis positive. All but three of these 151 mycobacterial isolates could be identified with ARDRA within on average 36 hours. The three isolates that could not be identified belonged to rare species not yet included in our ARDRA fingerprint library or were isolates with an aberrant pattern. CONCLUSIONS: In our hands, automated culture in combination with ARDRA provides with accurate, practically applicable, wide range identification of mycobacterial species. The existing identification library covers most species, and can be easily updated when new species are studied or described. The drawback is that ARDRA is culture-dependent, since automated culture of M. tuberculosis takes on average 16.7 days (range 6 to 29 days). However, culture is needed after all to assess the antibiotic susceptibility of the strains. BioMed Central 2002-03-01 /pmc/articles/PMC101405/ /pubmed/11945178 http://dx.doi.org/10.1186/1471-2180-2-4 Text en Copyright © 2002 De Baere et al; licensee BioMed Central Ltd. This is an Open Access article: verbatim copying and redistribution of this article are permitted in all media for any purpose, provided this notice is preserved along with the article's original URL.
spellingShingle Methodology Article
De Baere, Thierry
de Mendonça, Ricardo
Claeys, Geert
Verschraegen, Gerda
Mijs, Wouter
Verhelst, Rita
Rottiers, Sylvianne
Van Simaey, Leen
De Ganck, Catharine
Vaneechoutte, Mario
Evaluation of amplified rDNA restriction analysis (ARDRA) for the identification of cultured mycobacteria in a diagnostic laboratory
title Evaluation of amplified rDNA restriction analysis (ARDRA) for the identification of cultured mycobacteria in a diagnostic laboratory
title_full Evaluation of amplified rDNA restriction analysis (ARDRA) for the identification of cultured mycobacteria in a diagnostic laboratory
title_fullStr Evaluation of amplified rDNA restriction analysis (ARDRA) for the identification of cultured mycobacteria in a diagnostic laboratory
title_full_unstemmed Evaluation of amplified rDNA restriction analysis (ARDRA) for the identification of cultured mycobacteria in a diagnostic laboratory
title_short Evaluation of amplified rDNA restriction analysis (ARDRA) for the identification of cultured mycobacteria in a diagnostic laboratory
title_sort evaluation of amplified rdna restriction analysis (ardra) for the identification of cultured mycobacteria in a diagnostic laboratory
topic Methodology Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC101405/
https://www.ncbi.nlm.nih.gov/pubmed/11945178
http://dx.doi.org/10.1186/1471-2180-2-4
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