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Journal of Clinical Microbiology, July 1998, p. 1964-1968, Vol. 36, No. 7
0095-1137/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.

Comparison of the ABI 7700 System (TaqMan) and Competitive PCR for Quantification of IS6110 DNA in Sputum during Treatment of Tuberculosis

L. e. Desjardin,1 Y. Chen,1 M. D. Perkins,2 L. Teixeira,2 M. D. Cave,1,3 and K. D. Eisenach1,3,*

University of Arkansas for Medical Sciences1 and J. L. McClellan Memorial Veterans Affairs Hospital,3 Little Rock, Arkansas, and Duke University Medical Center/Universidade Federal do Espìrito Santo, Vitória, Brazil2

Received 21 January 1998/Returned for modification 9 March 1998/Accepted 28 March 1998

Mycobacterium tuberculosis can persist in sputum for long periods of time after the initiation of antituberculosis chemotherapy. The purpose of this study was to determine whether quantitative estimates of M. tuberculosis DNA in sputum correlate with the numbers of viable bacilli and thus measure the therapeutic response of patients during treatment. Two methods of M. tuberculosis DNA quantification were examined by using DNA isolated from sputum specimens serially collected during the course of chemotherapy. A competitive PCR assay was compared to an automated system of real-time quantification with the ABI Prism 7700 Sequence Detection System (TaqMan). The ABI 7700 system uses standard PCR in conjunction with a fluorogenic probe in which the intensity of fluorescence is proportional to the amount of target DNA present. The results showed that both PCR systems are reproducible and accurate. The amounts of M. tuberculosis DNA quantified in sputum corresponded well with the numbers of acid-fast bacilli (AFB) counted by microscopy. Before initiation of antituberculosis therapy, measures of AFB, M. tuberculosis DNA, and cultivable bacilli were similar, suggesting that quantification of DNA is a good method for measuring the initial bacillary load. However, the rate of disappearance of both AFB and M. tuberculosis DNA did not correlate with the decline in cultivable bacilli in the specimen; therefore, these tests are not appropriate for monitoring treatment efficacy.

* Corresponding author. Mailing address: Medical Research Service, Slot LR-151, J. L. McClellan Memorial VA Hospital, 4300 W. 7th St., Little Rock, AR 72205. Phone: (501) 660-2062. Fax: (501) 664-6748. E-mail: eisenachkathleend{at}exchange.uams.edu.

Journal of Clinical Microbiology, July 1998, p. 1964-1968, Vol. 36, No. 7
0095-1137/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.


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