title: Comparison of 14 molecular assays for detection of Mycobacterium tuberculosis complex in bronchoalveolar lavage fluid.
abstract: 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.
authors: University of Groningen, University Medical Center Groningen, Department of Pulmonology and Tuberculosis, Groningen, the Netherlands.
authors: Onno W
authors: OW
authors: Akkerman
authors: Tjip S
authors: TS
authors: van der Werf
authors: Maria
authors: M
authors: de Boer
authors: Jessica L
authors: JL
authors: de Beer
authors: Zeaur
authors: Z
authors: Rahim
authors: John W A
authors: JW
authors: Rossen
authors: Dick
authors: D
authors: van Soolingen
authors: Huib A M
authors: HA
authors: Kerstjens
authors: Adri G M
authors: AG
authors: van der Zanden
mesh_terms: D001431:Bacteriological Techniques / Q000379:methods*; D001992:Bronchoalveolar Lavage Fluid / Q000382:microbiology*; D006801:Humans; D025202:Molecular Diagnostic Techniques / Q000379:methods*; D009169:Mycobacterium tuberculosis / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification*; D012680:Sensitivity and Specificity; D014376:Tuberculosis / Q000175:diagnosis* / Q000382:microbiology
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