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

Vivek Kapur, Lingling Li, M. R. Hamrick, B. B. Plikaytis, T. M. Shinnick, A. Telenti, W. R. Jacobs, A. Banerjee, S. Cole, K. Y. Yuen, J. E. Clarridge, B. N. Kreiswirth, J. M. Musser

Research output: Contribution to journalArticle

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Abstract

Objective.-To develop and demonstrate the utility of automated DNA sequencing strategies for rapid and un-ambiguous 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 anti-microbial 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.

Original languageEnglish (US)
Pages (from-to)131-138
Number of pages8
JournalArchives of Pathology and Laboratory Medicine
Volume119
Issue number2
StatePublished - Jan 1 1995

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Mycobacterium
DNA Sequence Analysis
Mycobacterium tuberculosis
Tuberculosis
Sputum
Mutation
Genes
Heat-Shock Proteins
Base Pairing
Bacteria
Acids

All Science Journal Classification (ASJC) codes

  • Pathology and Forensic Medicine
  • Medical Laboratory Technology

Cite this

Kapur, Vivek ; Li, Lingling ; Hamrick, M. R. ; Plikaytis, B. B. ; Shinnick, T. M. ; Telenti, A. ; Jacobs, W. R. ; Banerjee, A. ; Cole, S. ; Yuen, K. Y. ; Clarridge, J. E. ; Kreiswirth, B. N. ; Musser, J. M. / Rapid Mycobacterium species assignment and unambiguous identification of mutations associated with antimicrobial resistance in Mycobacterium tuberculosis by automated DNA sequencing. In: Archives of Pathology and Laboratory Medicine. 1995 ; Vol. 119, No. 2. pp. 131-138.
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title = "Rapid Mycobacterium species assignment and unambiguous identification of mutations associated with antimicrobial resistance in Mycobacterium tuberculosis by automated DNA sequencing",
abstract = "Objective.-To develop and demonstrate the utility of automated DNA sequencing strategies for rapid and un-ambiguous 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 anti-microbial 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.",
author = "Vivek Kapur and Lingling Li and Hamrick, {M. R.} and Plikaytis, {B. B.} and Shinnick, {T. M.} and A. Telenti and Jacobs, {W. R.} and A. Banerjee and S. Cole and Yuen, {K. Y.} and Clarridge, {J. E.} and Kreiswirth, {B. N.} and Musser, {J. M.}",
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Kapur, V, Li, L, Hamrick, MR, Plikaytis, BB, Shinnick, TM, Telenti, A, Jacobs, WR, Banerjee, A, Cole, S, Yuen, KY, Clarridge, JE, Kreiswirth, BN & Musser, JM 1995, 'Rapid Mycobacterium species assignment and unambiguous identification of mutations associated with antimicrobial resistance in Mycobacterium tuberculosis by automated DNA sequencing', Archives of Pathology and Laboratory Medicine, vol. 119, no. 2, pp. 131-138.

Rapid Mycobacterium species assignment and unambiguous identification of mutations associated with antimicrobial resistance in Mycobacterium tuberculosis by automated DNA sequencing. / Kapur, Vivek; Li, Lingling; Hamrick, M. R.; Plikaytis, B. B.; Shinnick, T. M.; Telenti, A.; Jacobs, W. R.; Banerjee, A.; Cole, S.; Yuen, K. Y.; Clarridge, J. E.; Kreiswirth, B. N.; Musser, J. M.

In: Archives of Pathology and Laboratory Medicine, Vol. 119, No. 2, 01.01.1995, p. 131-138.

Research output: Contribution to journalArticle

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T1 - Rapid Mycobacterium species assignment and unambiguous identification of mutations associated with antimicrobial resistance in Mycobacterium tuberculosis by automated DNA sequencing

AU - Kapur, Vivek

AU - Li, Lingling

AU - Hamrick, M. R.

AU - Plikaytis, B. B.

AU - Shinnick, T. M.

AU - Telenti, A.

AU - Jacobs, W. R.

AU - Banerjee, A.

AU - Cole, S.

AU - Yuen, K. Y.

AU - Clarridge, J. E.

AU - Kreiswirth, B. N.

AU - Musser, J. M.

PY - 1995/1/1

Y1 - 1995/1/1

N2 - Objective.-To develop and demonstrate the utility of automated DNA sequencing strategies for rapid and un-ambiguous 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 anti-microbial 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.

AB - Objective.-To develop and demonstrate the utility of automated DNA sequencing strategies for rapid and un-ambiguous 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 anti-microbial 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.

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