Rapid Mycobacterium species assignment and unambiguous identification of mutations associated with antimicrobial resistance in Mycobacterium tuberculosis by automated DNA sequencing

Details

Serval ID
serval:BIB_D70B1425308C
Type
Article: article from journal or magazin.
Collection
Publications
Institution
Title
Rapid Mycobacterium species assignment and unambiguous identification of mutations associated with antimicrobial resistance in Mycobacterium tuberculosis by automated DNA sequencing
Journal
Archives of Pathology and Laboratory Medicine
Author(s)
Kapur  V., Li  L. L., Hamrick  M. R., Plikaytis  B. B., Shinnick  T. M., Telenti  A., Jacobs, W. R., Jr. , Banerjee  A., Cole  S., Yuen  K. Y., Clarridge  J.E., Kreiswirth  B.N., Musser  J.M.
ISSN
0003-9985 (Print)
Publication state
Published
Issued date
02/1995
Volume
119
Number
2
Pages
131-8
Notes
Journal Article Research Support, U.S. Gov't, P.H.S. --- Old month value: Feb
Abstract
OBJECTIVE--To develop and demonstrate the utility of automated DNA sequencing strategies for rapid and unambiguous identification of Mycobacterium species and mutations associated with antimicrobial resistance in Mycobacterium tuberculosis. DESIGN AND SPECIMENS--A 360-base pair segment of the gene (hsp65) encoding a 65-kd heat shock protein was characterized from 91 isolates assigned to 24 Mycobacterium species by traditional biochemical techniques. Areas of seven genes recently shown to contain mutations associated with antimicrobial resistance in M tuberculosis strains were also sequenced in a sample of 128 resistant organisms. Early positive BACTEC 460 cultures and acid-fast, bacterium-positive sputum specimens from patients with tuberculosis were also studied. RESULTS--Automated DNA sequencing identified species-specific polymorphism in the target segment of hsp65, successfully identified organisms to the species level in smear-positive sputum samples, and unambiguously characterized seven genes associated with antimicrobial resistance in M tuberculosis. CONCLUSIONS--Rapid identification of M tuberculosis and other Mycobacterium species is possible by automated DNA sequencing of a portion of hsp65. The technique is also feasible for analysis of some smear-positive sputum specimens. Unambiguous characterization of target segments of genes harboring mutations associated with antimicrobial resistance in M tuberculosis is possible from primary patient specimens. Taken together, the data demonstrate the feasibility of mycobacterial species identification and potential to identify mutations associated with antimicrobial resistance in less than 48 hours.
Keywords
Alleles Bacteriological Techniques Base Sequence Drug Resistance, Microbial/genetics Genes, Bacterial/*genetics Humans Molecular Sequence Data Mutation/*genetics Mycobacterium/genetics/*isolation & purification Mycobacterium tuberculosis/drug effects/genetics Sequence Analysis, DNA/*methods Sputum/microbiology
Pubmed
Web of science
Create date
25/01/2008 14:46
Last modification date
20/08/2019 15:56
Usage data