Rapid Identification of Mycobacteria to the Species Level Using INNO-LiPA Mycobacteria, a Reverse Hybridization Assay

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Published inJournal of Clinical Microbiology Vol. 39; no. 12; pp. 4477 - 4482
Main Authors SUFFYS, P. N, DA SILVA ROCHA, A, MIJS, W, VANDERBORGHT, B, DE OLIVEIRA, M, DIAS CAMPOS, C. E, BARRETO, A. M, PORTAELS, F, RIGOUTS, L, WOUTERS, G, JANNES, G, VAN REYBROECK, G
Format Journal Article
LanguageEnglish
Published Washington, DC American Society for Microbiology 01.12.2001
Subjects
Bacterial Proteins
Bacteriological methods and techniques used in bacteriology
Bacteriology
Biological and medical sciences
Chaperonin 60
Chaperonins - genetics
DNA Probes
DNA, Ribosomal Spacer - genetics
Fundamental and applied biological sciences. Psychology
Humans
Microbiology
Mycobacteriology and Aerobic Actinomycetes
Mycobacterium
Mycobacterium - classification
Mycobacterium - genetics
Mycobacterium Infections - microbiology
Mycological methods and techniques used in mycology
Mycology
Nucleic Acid Hybridization - methods
Polymerase Chain Reaction
Polymorphism, Restriction Fragment Length
Reagent Kits, Diagnostic
Reference Standards
Reproducibility of Results
reverse hybridization
RNA, Ribosomal, 16S - genetics
RNA, Ribosomal, 23S - genetics
Sensitivity and Specificity
Species Specificity
Time Factors
Mycobacteriaceae
Mycobacterium
Bacteria
Actinomycetes
Hybridization
Mycobacteriales
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Abstract Article Usage Stats Services JCM Citing Articles Google Scholar PubMed Related Content Social Bookmarking CiteULike Delicious Digg Facebook Google+ Mendeley Reddit StumbleUpon Twitter current issue JCM About JCM Subscribers Authors Reviewers Advertisers Inquiries from the Press Permissions & Commercial Reprints ASM Journals Public Access Policy JCM RSS Feeds 1752 N Street N.W. • Washington DC 20036 202.737.3600 • 202.942.9355 fax • journals@asmusa.org Print ISSN: 0095-1137 Online ISSN: 1098-660X Copyright © 2014 by the American Society for Microbiology.   For an alternate route to JCM .asm.org, visit: JCM       
AbstractList ABSTRACT INNO-LiPA Mycobacteria (LiPA; Innogenetics, Zwijnaarde, Belgium) is a kit for the simultaneous detection and identification of Mycobacterium species in culture and identifies the Mycobacterium tuberculosis complex, the M. avium complex (MAC), and the following Mycobacterium species: M. kansasii , M. avium , M. intracellulare , M. scrofulaceum , M. gordonae , M. xenopi , and the M. chelonae-M. abscessus complex. The assay, which targets the 16S-23S rRNA spacer region, was evaluated on 157 mycobacterial strains that had been identified by conventional techniques and PCR-restriction enzyme analysis of the hsp65 gene (PRA). Forty-seven reference strains consisting of 37 different species and 110 human clinical isolates were submitted to the test, and all were hybridized with the Mycobacterium genus probe (MYC) on the LiPA strip (100% sensitivity). Ninety-four isolates hybridized to their corresponding species- or complex-specific probes; only one isolate phenotypically identified as M. gordonae did not react with its specific probe (99.4% accuracy). Thirty-seven MAC strains were phenotypically identified to the complex level and to the species level by LiPA as M. avium ( n = 18) or M. intracellulare ( n = 7) or as belonging to the M. avium - M. intracellulare - M. scrofulaceum complex ( n = 12). Of the last 12 strains, 10 had M. avium PRA patterns and 2 had M. intracellulare PRA patterns. Three isolates that had been identified as a single species by conventional identification were proven to be mixed cultures by the LiPA assay. The whole procedure can be performed in 1 working day, starting with the supernatant of a small amount of bacterial mass that had been treated by freezing and then boiling.
Article Usage Stats Services JCM Citing Articles Google Scholar PubMed Related Content Social Bookmarking CiteULike Delicious Digg Facebook Google+ Mendeley Reddit StumbleUpon Twitter current issue JCM About JCM Subscribers Authors Reviewers Advertisers Inquiries from the Press Permissions & Commercial Reprints ASM Journals Public Access Policy JCM RSS Feeds 1752 N Street N.W. • Washington DC 20036 202.737.3600 • 202.942.9355 fax • journals@asmusa.org Print ISSN: 0095-1137 Online ISSN: 1098-660X Copyright © 2014 by the American Society for Microbiology.   For an alternate route to JCM .asm.org, visit: JCM       
INNO-LiPA Mycobacteria (LiPA; Innogenetics, Zwijnaarde, Belgium) is a kit for the simultaneous detection and identification of Mycobacterium species in culture and identifies the Mycobacterium tuberculosis complex, the M. avium complex (MAC), and the following Mycobacterium species: M. kansasii, M. avium, M. intracellulare, M. scrofulaceum, M. gordonae, M. xenopi, and the M. chelonae-M. abscessus complex. The assay, which targets the 16S-23S rRNA spacer region, was evaluated on 157 mycobacterial strains that had been identified by conventional techniques and PCR-restriction enzyme analysis of the hsp65 gene (PRA). Forty-seven reference strains consisting of 37 different species and 110 human clinical isolates were submitted to the test, and all were hybridized with the Mycobacterium genus probe (MYC) on the LiPA strip (100% sensitivity). Ninety-four isolates hybridized to their corresponding species- or complex-specific probes; only one isolate phenotypically identified as M. gordonae did not react with its specific probe (99.4% accuracy). Thirty-seven MAC strains were phenotypically identified to the complex level and to the species level by LiPA as M. avium (n = 18) or M. intracellulare (n = 7) or as belonging to the M. avium-M. intracellulare-M. scrofulaceum complex (n = 12). Of the last 12 strains, 10 had M. avium PRA patterns and 2 had M. intracellulare PRA patterns. Three isolates that had been identified as a single species by conventional identification were proven to be mixed cultures by the LiPA assay. The whole procedure can be performed in 1 working day, starting with the supernatant of a small amount of bacterial mass that had been treated by freezing and then boiling.
INNO-LiPA Mycobacteria (LiPA; Innogenetics, Zwijnaarde, Belgium) is a kit for the simultaneous detection and identification of Mycobacterium species in culture and identifies the Mycobacterium tuberculosis complex, the M. avium complex (MAC), and the following Mycobacterium species: M. kansasii , M. avium , M. intracellulare , M. scrofulaceum , M. gordonae , M. xenopi , and the M. chelonae-M. abscessus complex. The assay, which targets the 16S-23S rRNA spacer region, was evaluated on 157 mycobacterial strains that had been identified by conventional techniques and PCR-restriction enzyme analysis of the hsp65 gene (PRA). Forty-seven reference strains consisting of 37 different species and 110 human clinical isolates were submitted to the test, and all were hybridized with the Mycobacterium genus probe (MYC) on the LiPA strip (100% sensitivity). Ninety-four isolates hybridized to their corresponding species- or complex-specific probes; only one isolate phenotypically identified as M. gordonae did not react with its specific probe (99.4% accuracy). Thirty-seven MAC strains were phenotypically identified to the complex level and to the species level by LiPA as M. avium ( n = 18) or M. intracellulare ( n = 7) or as belonging to the M. avium - M. intracellulare - M. scrofulaceum complex ( n = 12). Of the last 12 strains, 10 had M. avium PRA patterns and 2 had M. intracellulare PRA patterns. Three isolates that had been identified as a single species by conventional identification were proven to be mixed cultures by the LiPA assay. The whole procedure can be performed in 1 working day, starting with the supernatant of a small amount of bacterial mass that had been treated by freezing and then boiling.
Author G. van Reybroeck
C. E. Dias Campos
B. Vanderborght
M. de Oliveira
F. Portaels
G. Jannes
G. Wouters
W. Mijs
L. Rigouts
P. N. Suffys
A. M. Werneck Barreto
A. da Silva Rocha
AuthorAffiliation Biochemistry and Molecular Biology Department, Oswaldo Cruz Institute, Oswaldo Cruz Foundation, 1 Reference Center Professor Hélio Fraga, 2 and Laboratory of Molecular Biology, HUCFF, Federal University of Rio de Janeiro, 5 Rio de Janeiro, Brazil, and Institute of Tropical Medicine, Antwerp, 3 and Innogenetics, Zwijnaarde, 4 Belgium
AuthorAffiliation_xml – name: Biochemistry and Molecular Biology Department, Oswaldo Cruz Institute, Oswaldo Cruz Foundation, 1 Reference Center Professor Hélio Fraga, 2 and Laboratory of Molecular Biology, HUCFF, Federal University of Rio de Janeiro, 5 Rio de Janeiro, Brazil, and Institute of Tropical Medicine, Antwerp, 3 and Innogenetics, Zwijnaarde, 4 Belgium
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Corresponding author. Mailing address: Laboratory of Molecular and Diagnosis of Infectious Diseases, DBBM, IOC, Fiocruz, Av. Brasil 4365, Manguinhos 21045-900, Rio de Janeiro, Brazil. Phone: 55-21-5984289. Fax: 55-21-2709997. E-mail: psuffys@ioc.fiocruz.br.
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SSID ssj0014455
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Snippet Article Usage Stats Services JCM Citing Articles Google Scholar PubMed Related Content Social Bookmarking CiteULike Delicious Digg Facebook Google+ Mendeley...
INNO-LiPA Mycobacteria (LiPA; Innogenetics, Zwijnaarde, Belgium) is a kit for the simultaneous detection and identification of Mycobacterium species in culture...
ABSTRACT INNO-LiPA Mycobacteria (LiPA; Innogenetics, Zwijnaarde, Belgium) is a kit for the simultaneous detection and identification of Mycobacterium species...
INNO-LiPA Mycobacteria (LiPA; Innogenetics, Zwijnaarde, Belgium) is a kit for the simultaneous detection and identification of Mycobacterium species in culture...
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SourceType Open Access Repository
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StartPage 4477
SubjectTerms Bacterial Proteins
Bacteriological methods and techniques used in bacteriology
Bacteriology
Biological and medical sciences
Chaperonin 60
Chaperonins - genetics
DNA Probes
DNA, Ribosomal Spacer - genetics
Fundamental and applied biological sciences. Psychology
Humans
Microbiology
Mycobacteriology and Aerobic Actinomycetes
Mycobacterium
Mycobacterium - classification
Mycobacterium - genetics
Mycobacterium Infections - microbiology
Mycological methods and techniques used in mycology
Mycology
Nucleic Acid Hybridization - methods
Polymerase Chain Reaction
Polymorphism, Restriction Fragment Length
Reagent Kits, Diagnostic
Reference Standards
Reproducibility of Results
reverse hybridization
RNA, Ribosomal, 16S - genetics
RNA, Ribosomal, 23S - genetics
Sensitivity and Specificity
Species Specificity
Time Factors
Title Rapid Identification of Mycobacteria to the Species Level Using INNO-LiPA Mycobacteria, a Reverse Hybridization Assay
URI http://jcm.asm.org/content/39/12/4477.abstract
https://www.ncbi.nlm.nih.gov/pubmed/11724865
https://search.proquest.com/docview/18218554
https://search.proquest.com/docview/72398442
https://pubmed.ncbi.nlm.nih.gov/PMC88569
Volume 39
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