Measurement of sputum Mycobacterium tuberculosis messenger RNA as a surrogate for response to chemotherapy

Lucy E. Desjardin, Mark D. Perkins, Kathy Wolski, Shirley Haun, Luciléia Teixeira, Ying Chen, John L. Johnson, Jerrold J. Ellner, Reynaldo Dietze, Joseph Bates, M. Donald Cave, Kathleen D. Eisenach

Research output: Contribution to journalArticle

Abstract

Effective treatment regimens for pulmonary tuberculosis are difficult to assess because of the slow growth rate of Mycobacterium tuberculosis in culture and its protracted clearance from sputum. A rapid method that reflects effective antimicrobial activity would markedly advance evaluation of treatment and promote the assessment of new antituberculosis drugs. Conventional methods measure the progressive reduction of numbers of acid- fast bacilli in the sputum smear and the clearance of organisms in sputum culture. In this study, we measured levels of M. tuberculosis 85B (alpha antigen) messenger RNA (mRNA), 16S ribosomal RNA (rRNA), and IS6110 DNA in patients' sputa to ascertain whether they could serve as potential surrogate markers of response to chemotherapy. Sputum specimens were sequentially collected for up to a year from 19 smear-positive pulmonary tuberculosis patients receiving an optimal drug treatment regimen. Nucleic acids were isolated from these specimens, and two M. tuberculosis molecular targets (mRNA, DNA) were quantified, using the ABI Prism 7700 Sequence Detection System. The Mycobacterium genus-specific 16S rRNA was quantified with a limiting dilution RT-PCR assay. Results show that levels of 85B mRNA declined after initiation of therapy, as did viable M. tuberculosis colony counts, with 90% of patients becoming negative for both markers after 2 mo of treatment. The rapid disappearance of M. tuberculosis mRNA from sputum suggests that it is a good indicator of microbial viability and a useful marker for rapid assessment of response to chemotherapy.

Original languageEnglish (US)
Pages (from-to)203-210
Number of pages8
JournalAmerican Journal of Respiratory and Critical Care Medicine
Volume160
Issue number1
StatePublished - 1999
Externally publishedYes

Fingerprint

Sputum
Mycobacterium tuberculosis
Drug Therapy
Pulmonary Tuberculosis
Messenger RNA
16S Ribosomal RNA
Microbial Viability
Therapeutics
Mycobacterium
Ribosomal DNA
Pharmaceutical Preparations
Nucleic Acids
Bacillus
Biomarkers
Polymerase Chain Reaction
Acids
DNA
Growth

ASJC Scopus subject areas

  • Pulmonary and Respiratory Medicine

Cite this

Desjardin, L. E., Perkins, M. D., Wolski, K., Haun, S., Teixeira, L., Chen, Y., ... Eisenach, K. D. (1999). Measurement of sputum Mycobacterium tuberculosis messenger RNA as a surrogate for response to chemotherapy. American Journal of Respiratory and Critical Care Medicine, 160(1), 203-210.

Measurement of sputum Mycobacterium tuberculosis messenger RNA as a surrogate for response to chemotherapy. / Desjardin, Lucy E.; Perkins, Mark D.; Wolski, Kathy; Haun, Shirley; Teixeira, Luciléia; Chen, Ying; Johnson, John L.; Ellner, Jerrold J.; Dietze, Reynaldo; Bates, Joseph; Cave, M. Donald; Eisenach, Kathleen D.

In: American Journal of Respiratory and Critical Care Medicine, Vol. 160, No. 1, 1999, p. 203-210.

Research output: Contribution to journalArticle

Desjardin, LE, Perkins, MD, Wolski, K, Haun, S, Teixeira, L, Chen, Y, Johnson, JL, Ellner, JJ, Dietze, R, Bates, J, Cave, MD & Eisenach, KD 1999, 'Measurement of sputum Mycobacterium tuberculosis messenger RNA as a surrogate for response to chemotherapy', American Journal of Respiratory and Critical Care Medicine, vol. 160, no. 1, pp. 203-210.
Desjardin, Lucy E. ; Perkins, Mark D. ; Wolski, Kathy ; Haun, Shirley ; Teixeira, Luciléia ; Chen, Ying ; Johnson, John L. ; Ellner, Jerrold J. ; Dietze, Reynaldo ; Bates, Joseph ; Cave, M. Donald ; Eisenach, Kathleen D. / Measurement of sputum Mycobacterium tuberculosis messenger RNA as a surrogate for response to chemotherapy. In: American Journal of Respiratory and Critical Care Medicine. 1999 ; Vol. 160, No. 1. pp. 203-210.
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