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首页> 外文期刊>Journal of Clinical Microbiology >Comparison of the ABI 7700 System (TaqMan) and Competitive PCR for Quantification of IS6110 DNA in Sputum during Treatment of Tuberculosis
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Comparison of the ABI 7700 System (TaqMan) and Competitive PCR for Quantification of IS6110 DNA in Sputum during Treatment of Tuberculosis

机译：ABI 7700系统（TaqMan）和竞争性PCR定量分析结核病期间痰液中IS6110 DNA的比较

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摘要

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.
机译：抗结核化疗开始后，结核分枝杆菌可以长期存在于痰中。这项研究的目的是确定是否定量估计 M。痰中的结核DNA与活菌数有关，从而可以衡量患者在治疗过程中的治疗反应。 M的两种方法。通过从化疗过程中连续收集的痰标本中分离出的DNA来检查结核病的DNA定量。使用ABI Prism 7700序列检测系统（TaqMan）将竞争性PCR分析与实时定量自动化系统进行了比较。 ABI 7700系统将标准PCR与荧光探针结合使用，荧光强度与存在的目标DNA的数量成正比。结果表明，两种PCR系统均具有可重复性和准确性。 M的数量。痰中检出的肺结核DNA与显微镜下计数的抗酸杆菌数（AFB）吻合得很好。在开始抗结核治疗之前，应先测量AFB， M。结核DNA和可培养的细菌相似，这表明DNA定量是测量初始细菌载量的好方法。但是，AFB和 M的消失率都很高。结核DNA与标本中可培养细菌的减少无关。因此，这些测试不适用于监测治疗效果。

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