hsp65 sequencing for identification of rapidly growing mycobacteria.

Journal of clinical microbiology ·Vol. 37 ·No. 3 ·1999-03-00 ·Pages 852-7

Ringuet H, Akoua-Koffi C, Honore S, Varnerot A, Vincent V, Berche P, Gaillard JL, Pierre-Audigier C

Abstract

Partial sequencing of the hsp65 gene was used for the identification of rapidly growing mycobacteria (RGM). A 441-bp fragment (A. Telenti, F. Marchesi, M. Balz, F. Bally, E. Böttger, and T. Bodmer, J. Clin. Microbiol. 31:175-178, 1993) was amplified and sequenced by an automated fluorescence-based method involving capillary electrophoresis. Type strains of 10 RGM species were first studied. Each species had a unique nucleotide sequence, distinguishing it clearly from the other species. A panel of strains from the four main RGM species responsible for human infections, Mycobacterium abscessus, Mycobacterium chelonae, Mycobacterium fortuitum, and Mycobacterium peregrinum, was also studied. There were few sequence differences within each of these species (<2% of bases were different from the type strain sequence), and they had no effect on species assignment. hsp65 sequencing unambiguously differentiated M. chelonae and M. abscessus, two species difficult to identify by classical methods and 16S rRNA gene sequencing. The devised procedure is a rapid and reliable tool for the identification of RGM species.

MeSH Terms

Antigens, Bacterial/genetics Bacterial Proteins Base Sequence Chaperonin 60 Chaperonins/genetics Genetic Variation Humans Molecular Sequence Data Mycobacterium/classification,genetics,growth & development Mycobacterium Infections/diagnosis Mycobacterium tuberculosis/genetics,growth & development Phylogeny Polymerase Chain Reaction/methods RNA, Bacterial/genetics RNA, Ribosomal, 16S/genetics Sequence Alignment Sequence Homology, Nucleic Acid

Chemicals

Antigens, Bacterial Bacterial Proteins Chaperonin 60 RNA, Bacterial RNA, Ribosomal, 16S heat-shock protein 65, Mycobacterium Chaperonins

Authors & Affiliations

8 authors, click to expand affiliations / ORCID

Ringuet H

Service de Microbiologie, Hôpital Necker-Enfants Malades, 75015 Paris, France.

Akoua-Koffi C

Honore S

Varnerot A

Vincent V

Berche P

Gaillard J L

Pierre-Audigier C

References (26)

26 references, click to expand

Genetic heterogeneity within Mycobacterium fortuitum complex species: genotypic criteria for identification.
J Clin Microbiol. 1992 Nov;30(11):2772-5 PMID: 1280641
Differentiation of slowly growing Mycobacterium species, including Mycobacterium tuberculosis, by gene amplification and restriction fragment length polymorphism analysis.
J Clin Microbiol. 1992 Jul;30(7):1815-22 PMID: 1352786
Proposal of Mycobacterium peregrinum sp. nov., nom. rev., and elevation of Mycobacterium chelonae subsp. abscessus (Kubica et al.) to species status: Mycobacterium abscessus comb. nov.
Int J Syst Bacteriol. 1992 Apr;42(2):240-5 PMID: 1581184
Detection and identification of mycobacteria by amplification of mycobacterial DNA.
Mol Microbiol. 1989 Jul;3(7):843-9 PMID: 2507865
The 65-kilodalton antigen of Mycobacterium tuberculosis.
J Bacteriol. 1987 Mar;169(3):1080-8 PMID: 3029018
Genotypic identification of mycobacteria by nucleic acid sequence determination: report of a 2-year experience in a clinical laboratory.
J Clin Microbiol. 1993 Nov;31(11):2882-9 PMID: 7505291
Mycobacterial taxonomy.
Eur J Clin Microbiol Infect Dis. 1994 Nov;13(11):884-901 PMID: 7698114
Recent changes in taxonomy and disease manifestations of the rapidly growing mycobacteria.
Eur J Clin Microbiol Infect Dis. 1994 Nov;13(11):953-60 PMID: 7698122
PCR amplification and restriction endonuclease analysis of a 65-kilodalton heat shock protein gene sequence for taxonomic separation of rapidly growing mycobacteria.
J Clin Microbiol. 1995 Jan;33(1):149-53 PMID: 7699032
Rapid Mycobacterium species assignment and unambiguous identification of mutations associated with antimicrobial resistance in Mycobacterium tuberculosis by automated DNA sequencing.
Arch Pathol Lab Med. 1995 Feb;119(2):131-8 PMID: 7848059
The superoxide dismutase gene, a target for detection and identification of mycobacteria by PCR.
J Clin Microbiol. 1994 Nov;32(11):2801-12 PMID: 7852575
Nucleotide sequence comparison of the mycobacterial dnaJ gene and PCR-restriction fragment length polymorphism analysis for identification of mycobacterial species.
Int J Syst Bacteriol. 1994 Jan;44(1):159-66 PMID: 7907223
DNA large restriction fragment patterns of sporadic and epidemic nosocomial strains of Mycobacterium chelonae and Mycobacterium abscessus.
J Clin Microbiol. 1993 Oct;31(10):2697-701 PMID: 8253968
Rapid identification of mycobacteria to the species level by polymerase chain reaction and restriction enzyme analysis.
J Clin Microbiol. 1993 Feb;31(2):175-8 PMID: 8381805
Identification of Mycobacterium species by using amplified ribosomal DNA restriction analysis.
J Clin Microbiol. 1993 Aug;31(8):2061-5 PMID: 8396586
Disseminated infection with rapidly growing mycobacteria.
Clin Infect Dis. 1993 Apr;16(4):463-71 PMID: 8513048
Two-laboratory collaborative study on identification of mycobacteria: molecular versus phenotypic methods.
J Clin Microbiol. 1996 Feb;34(2):296-303 PMID: 8789004
Identification and subspecific differentiation of Mycobacterium scrofulaceum by automated sequencing of a region of the gene (hsp65) encoding a 65-kilodalton heat shock protein.
J Clin Microbiol. 1996 Dec;34(12):3151-9 PMID: 8940463
Routine use of PCR-restriction fragment length polymorphism analysis for identification of mycobacteria growing in liquid media.
J Clin Microbiol. 1997 Jan;35(1):79-85 PMID: 8968884
Diversity of the 32-kilodalton protein gene may form a basis for species determination of potentially pathogenic mycobacterial species.
J Clin Microbiol. 1997 Mar;35(3):769-73 PMID: 9041432
Subspecific differentiation of Mycobacterium avium complex strains by automated sequencing of a region of the gene (hsp65) encoding a 65-kilodalton heat shock protein.
Int J Syst Bacteriol. 1997 Apr;47(2):414-9 PMID: 9103630
PCR-enzyme-linked immunosorbent assay and partial rRNA gene sequencing: a rational approach to identifying mycobacteria.
J Clin Microbiol. 1997 Sep;35(9):2375-80 PMID: 9276419
Routine rapid Mycobacterium species assignment based on species-specific allelic variation in the 65-kilodalton heat shock protein gene (hsp65).
Arch Pathol Lab Med. 1997 Aug;121(8):859-64 PMID: 9278615
Rapid identification of mycobacteria to species level by PCR-restriction fragment length polymorphism analysis of the hsp65 gene and proposition of an algorithm to differentiate 34 mycobacterial species.
J Clin Microbiol. 1997 Nov;35(11):2969-73 PMID: 9350770
Differentiation of phylogenetically related slowly growing mycobacteria based on 16S-23S rRNA gene internal transcribed spacer sequences.
J Clin Microbiol. 1998 Jan;36(1):139-47 PMID: 9431937
Clinical application of PCR-restriction enzyme pattern analysis for rapid identification of aerobic actinomycete isolates.
J Clin Microbiol. 1998 Jan;36(1):148-52 PMID: 9431938

Article Info

Journal

Journal of clinical microbiology

Abbr.

J Clin Microbiol

ISSN

0095-1137

DOI

10.1128/JCM.37.3.852-857.1999

Published

1999-03-00

Pages

852-7

Language

English

Region

United States

NLM ID

7505564

PMCID

PMC84584

Subset

Databases

GENBANK

AF071128, AF071129, AF071130, AF071131, AF071132, AF071133, AF071134, AF071135, AF071136, AF071137, AF071138, AF071139, AF071140, AF071141, AF071142, AF140676, AF140677

External Links

PubMed source DOI: 10.1128/JCM.37.3.852-857.1999 PMC Full Text View journal details

Analysis Services

hsp65 sequencing for identification of rapidly growing mycobacteria.

Contact

WeChat Official Account