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Comparison of 14 Molecular Assays for Detection of Mycobacterium tuberculosis Complex in Bronchoalveolar Lavage Fluid

机译:检测支气管肺泡灌洗液中结核分枝杆菌复合物的14种分子检测方法的比较

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We compared 14 molecular assays for their ability to detect the Mycobacterium tuberculosis complex in bronchoalveolar lavage fluid samples. Three approaches were followed. First, by using DNA from Mycobacterium bovis BCG, we determined the detection limits of the assays using routine molecular methods. Second, in order to determine the analytical sensitivities of the assays, we added one of four M. tuberculosis isolates with various numbers of the insertion sequence IS6110 to N-acetyl-l-cysteine (NALC)-NaOH-treated bronchoalveolar lavage fluid samples in dilutions of 1:10 to 1:10,000,000. Third, intertest variabilities were measured and defined by the standard deviations for the quantitation cycle (Cq) values of three positive test results per dilution per assay. The 14 assays tested had similar analytical sensitivities, except for GeneXpert, which had an analytical sensitivity that was 10- to 100-fold lower than that of the other assays. The MP MTB/NTM test and the in-house TaqMan-10 revealed the best performances for the detection limit and had the highest analytical sensitivities. Most of the tests performed well regarding detection limit and analytical sensitivity for the detection of the M. tuberculosis complex in serial dilutions, and the differences were small. The MP MTB/NTM and the in-house TaqMan-10 assays revealed the best, and GeneXpert the worst, overall performances.
机译:我们比较了14种分子测定法检测支气管肺泡灌洗液样本中结核分枝杆菌复合物的能力。遵循了三种方法。首先,通过使用牛分枝杆菌BCG的DNA,我们使用常规分子方法确定了检测的检测限。其次,为了确定测定的分析灵敏度,我们将四种具有不同插入序列IS 6110 插入序列的结核分枝杆菌菌株之一添加到 N -乙酰基-用L-半胱氨酸(NALC)-NaOH处理过的支气管肺泡灌洗液样品,稀释比例为1:10至1:10,000,000。第三,测试间的变异性由每个测定的每个稀释度的三个阳性测试结果的定量循环(Cq)值的标准偏差测量并定义。除了GeneXpert,GeneXpert的分析灵敏度比其他分析低10到100倍,测试的14种测定具有相似的分析灵敏度。 MP MTB / NTM测试和内部TaqMan-10显示出最佳的检测限性能,并具有最高的分析灵敏度。大多数测试在连续稀释液中检测结核分枝杆菌复合物的检测限和分析灵敏度方面都表现良好,差异很小。 MP MTB / NTM和内部TaqMan-10检测显示出最佳的整体性能,而GeneXpert则显示出最差的整体性能。

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