Rapid, standardized method for determination of Mycobacterium tuberculosis drug susceptibility by use of mycolic acid analysis

Nicole M Parrish, Gerard Osterhout, Kim Dionne, Amy Sweeney, Nicole Kwiatkowski, Karen C Carroll, Kenneth C. Jost, James Dick

Research output: Contribution to journalArticle

Abstract

Multidrug-resistant (MDR) Mycobacterium tuberculosis and extrensively drug-resistant (XDR) M. tuberculosis are emerging public health threats whose threats are compounded by the fact that current techniques for testing the susceptibility of M. tuberculosis require several days to weeks to complete. We investigated the use of high-performance liquid chromatography (HPLC)-based quantitation of mycolic acids as a means of rapidly determining drug resistance and susceptibility in M. tuberculosis. Standard susceptibility testing and determination of the MICs of drug-susceptible (n = 26) and drug-resistant M. tuberculosis strains, including MDR M. tuberculosis strains (n = 34), were performed by using the Bactec radiometric growth system as the reference method. The HPLC-based susceptibilities of the current first-line drugs, isoniazid (INH), rifampin (RIF), ethambutol (EMB), and pyrazinamide (PZA), were determined. The vials were incubated for 72 h, and aliquots were removed for HPLC analysis by using the Sherlock mycobacterial identification system. HPLC quantitation of total mycolic acid peaks (TMAPs) was performed with treated and untreated cultures. At 72 h, the levels of agreement of the HPLC method with the reference method were 99.5% for INH, EMB, and PZA and 98.7% for RIF. The inter- and intra-assay reproducibilities varied by drug, with an average precision of 13.4%. In summary, quantitation of TMAPs is a rapid, sensitive, and accurate method for antibiotic susceptibility testing of all first-line drugs currently used against M. tuberculosis and offers the potential of providing susceptibility testing results within hours, rather than days or weeks, for clinical M. tuberculosis isolates.

Original languageEnglish (US)
Pages (from-to)3915-3920
Number of pages6
JournalJournal of Clinical Microbiology
Volume45
Issue number12
DOIs
StatePublished - Dec 2007

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Mycolic Acids
Mycobacterium tuberculosis
Multidrug-Resistant Tuberculosis
High Pressure Liquid Chromatography
Pharmaceutical Preparations
Pyrazinamide
Ethambutol
Rifampin
Mycobacterium
Isoniazid
Drug Resistance
Public Health
Anti-Bacterial Agents
Growth

ASJC Scopus subject areas

  • Microbiology (medical)
  • Microbiology

Cite this

Rapid, standardized method for determination of Mycobacterium tuberculosis drug susceptibility by use of mycolic acid analysis. / Parrish, Nicole M; Osterhout, Gerard; Dionne, Kim; Sweeney, Amy; Kwiatkowski, Nicole; Carroll, Karen C; Jost, Kenneth C.; Dick, James.

In: Journal of Clinical Microbiology, Vol. 45, No. 12, 12.2007, p. 3915-3920.

Research output: Contribution to journalArticle

Parrish, Nicole M ; Osterhout, Gerard ; Dionne, Kim ; Sweeney, Amy ; Kwiatkowski, Nicole ; Carroll, Karen C ; Jost, Kenneth C. ; Dick, James. / Rapid, standardized method for determination of Mycobacterium tuberculosis drug susceptibility by use of mycolic acid analysis. In: Journal of Clinical Microbiology. 2007 ; Vol. 45, No. 12. pp. 3915-3920.
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