title: Comparison of the ABI 7700 system (TaqMan) and competitive PCR for quantification of IS6110 DNA in sputum during treatment of tuberculosis.
abstract: 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.
authors.affiliation: University of Arkansas for Medical Sciences, Little Rock, USA.
authors.forename: L E
authors.initials: LE
authors.lastname: Desjardin
authors.forename: Y
authors.initials: Y
authors.lastname: Chen
authors.forename: M D
authors.initials: MD
authors.lastname: Perkins
authors.forename: L
authors.initials: L
authors.lastname: Teixeira
authors.forename: M D
authors.initials: MD
authors.lastname: Cave
authors.forename: K D
authors.initials: KD
authors.lastname: Eisenach
mesh_terms: D000995:Antitubercular Agents / Q000627:therapeutic use; D001431:Bacteriological Techniques; D001483:Base Sequence; D001667:Binding, Competitive; D015169:Colony Count, Microbial; D017931:DNA Primers; D004251:DNA Transposable Elements; D004269:DNA, Bacterial / Q000032:analysis; D004359:Drug Therapy, Combination; D006801:Humans; D008969:Molecular Sequence Data; D009169:Mycobacterium tuberculosis / Q000235:genetics / Q000302:isolation & purification*; D016133:Polymerase Chain Reaction / Q000379:methods*; D013183:Sputum / Q000382:microbiology*; D013997:Time Factors; D014397:Tuberculosis, Pulmonary / Q000188:drug therapy* / Q000382:microbiology*
chemical_list: D000995:Antitubercular Agents; D017931:DNA Primers; D004251:DNA Transposable Elements; D004269:DNA, Bacterial