Home LiteratureArticle Details
PMID: 10325367 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Species-specific identification of Mycobacterium leprae by PCR-restriction fragment length polymorphism analysis of the hsp65 gene.

Journal of clinical microbiology ·Vol. 37 ·No. 6 ·1999-06-00 ·Pages 2016-9

Rastogi N, Goh KS, Berchel M

Abstract

PCR-restriction fragment length polymorphism analysis (PRA) of the hsp65 gene present in all mycobacteria was used in the present investigation to characterize Mycobacterium leprae. Bacilli were extracted and purified from different organs from experimentally infected armadillos and nude mice (Swiss mice of nu/nu origin). A total of 15 samples were assayed in duplicate, and the results were compared with those obtained for a total of 147 cultivable mycobacteria representing 34 species. Irrespective of its origin or viability, M. leprae strains from all the samples were uniformly characterized by two fragments of 315 and 135 bp upon BstEII digestion and two fragments of 265 and 130 bp upon HaeIII digestion. PRA is a relatively simple method and permits the conclusive identification of M. leprae to the species level.

MeSH Terms
Algorithms Animals Antigens, Bacterial/genetics Armadillos Bacterial Proteins Chaperonin 60 Chaperonins/genetics Genes, Bacterial Leprosy/diagnosis Liver/microbiology Lymph Nodes/microbiology Mice Mice, Nude Mycobacterium leprae/genetics,isolation & purification Mycobacterium tuberculosis/genetics,isolation & purification Polymerase Chain Reaction/methods Polymorphism, Restriction Fragment Length Sensitivity and Specificity Species Specificity Spleen/microbiology
Chemicals
Antigens, Bacterial Bacterial Proteins Chaperonin 60 heat-shock protein 65, Mycobacterium Chaperonins
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Rastogi N
Unité de la Tuberculose et des Mycobactéries, Institut Pasteur, 97165 Pointe à Pitre Cedex, Guadeloupe. [email protected]
Goh K S
Berchel M
References (15)
15 references, click to expand
  1. Efficient mapping of protein antigenic determinants.
    Proc Natl Acad Sci U S A. 1986 Sep;83(18):7013-7 PMID: 2428046
  2. Changes in the morphology of Mycobacterium leprae in patients under treatment.
    Int J Lepr. 1962 Jul-Sep;30:266-77 PMID: 13999150
  3. Mycobacterium leprae surface components intervene in the early phagosome-lysosome fusion inhibition event.
    Infect Immun. 1987 Dec;55(12):2916-21 PMID: 3316028
  4. The Mycobacterium tuberculosis 65-kilodalton antigen is a heat shock protein which corresponds to common antigen and to the Escherichia coli GroEL protein.
    Infect Immun. 1988 Feb;56(2):446-51 PMID: 2892795
  5. Rapid and sensitive detection of Mycobacterium leprae using a nested-primer gene amplification assay.
    J Clin Microbiol. 1990 Sep;28(9):1913-7 PMID: 2229372
  6. Differential handling of bacterial antigens in macrophages infected with Mycobacterium leprae as studied by immunogold labeling of ultrathin sections.
    Int J Lepr Other Mycobact Dis. 1991 Jun;59(2):278-91 PMID: 2071985
  7. Species-specific assessment of Mycobacterium leprae in skin biopsies by in situ hybridization and polymerase chain reaction.
    Lab Invest. 1992 May;66(5):618-23 PMID: 1573855
  8. 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
  9. Evaluation of polymerase chain reaction amplification of Mycobacterium leprae-specific repetitive sequence in biopsy specimens from leprosy patients.
    J Clin Microbiol. 1993 Apr;31(4):895-9 PMID: 8463401
  10. 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
  11. Bangkok Workshop on Leprosy Research. Diagnosis, classification and prognosis.
    Int J Lepr Other Mycobact Dis. 1996 Dec;64(4 Suppl):S13-4; discussion S14-5 PMID: 9044915
  12. 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
  13. Reverse transcription-PCR detection of Mycobacterium leprae in clinical specimens.
    J Clin Microbiol. 1998 May;36(5):1352-6 PMID: 9574704
  14. Polymerase chain reaction assay for the detection and identification of Mycobacterium leprae in patients in the United States.
    Am J Clin Pathol. 1998 May;109(5):642-6 PMID: 9576586
  15. The 65-kilodalton antigen of Mycobacterium tuberculosis.
    J Bacteriol. 1987 Mar;169(3):1080-8 PMID: 3029018
Article Info
Journal
Journal of clinical microbiology
Abbr.
J Clin Microbiol
ISSN
0095-1137
DOI
10.1128/JCM.37.6.2016-2019.1999
Email
[email protected]
Published
1999-06-00
Pages
2016-9
Language
English
Region
United States
NLM ID
7505564
PMCID
PMC85014
Subset
IM
External Links
PubMed source DOI: 10.1128/JCM.37.6.2016-2019.1999 PMC Full Text View journal details
Analysis Services
Analysis Services

Contact

No. 2 Wenbo Road, Zhangqiu District, Jinan, Shandong

Qilu Normal University · Genelibs Bioinformatics Lab

750 Shunhua Rd, Jinan

2F, Bldg F, University Science Park

Tel: 0531-88819269

WeChat Official Account

Follow our WeChat subscription account for real-time updates and the latest in medical and biological research.


Business Email

E-mail: [email protected]

Copyright © 2026.Genelibs Jinan Honghui Biotechnology Co., Ltd. 鲁ICP备20007159号-2