One obstacle to wider use of rapid liquid culture-based tuberculosis diagnostics such as the microscopic observation drug susceptibility (MODS) assay is concern about cross-contamination. We investigated the rate of laboratory cross-contamination in MODS, automated MBBacT, and Lowenstein-Jensen (LJ) cultures performed in parallel, through triangulation of microbiologic (reculturing stored samples), molecular (spoligotype/RFLP), and clinical epidemiologic data. At least 1 culture was positive for Mycobacterium tuberculosis for 362 (11%) of 3416 samples; 53 were regarded as potential cross-contamination suspects. Cross-contamination accounted for 17 false-positive cultures from 14 samples representing 0.41% (14/3416) and 0.17% (17/10 248) of samples and cultures, respectively. Positive predictive values for MODS, MBBacT (bioMérieux, Durham, NC), and LJ were 99.1%, 98.7%, and 99.7%, and specificity was 99.9% for all 3. Low rates of cross-contamination are achievable in mycobacterial laboratories in resource-poor settings even when a large proportion of samples are infectious and highly sensitive liquid culture-based diagnostics such as MODS are used.
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BackgroundIn mycobacteriology reference-level laboratories in the industrialized world, crosscontamination is estimated to account for between 0.5% and 6% of positive results (Bauer et al., 1997;Nivin et al., 1998;Burman and Reves 2000;de Boer et al., 2002;Jasmer et al., 2002;Ruddy et al., 2002) with significant associated cost implications (Northrup et al., 2002), and higher levels might be anticipated in high tuberculosis (TB)-burden resourcelimited settings where laboratory facilities are less sophisticated and a greater proportion of samples contain Mycobacterium tuberculosis. The microscopic observation drug susceptibility (MODS) assay in which 2 sputum samples are cultured on the same 24-well tissue-culture plate in Middlebrook 7H9 medium (Caviedes et al., 2000;Moore et al., 2004) has been proposed as a potential tool to bring low-tech sensitive TB diagnosis to the developing world where the need is most urgent; however, the risk of cross-contamination in this simple low-tech method using liquid culture medium and multiple samples in a single 24-well plate has not been previously determined.Examining samples obtained in a large community-based study in urban Lima, Peru, we determined the rate of M. tuberculosis cross-contamination in sputum cultures performed in parallel in MODS, MBBacT-automated mycobacterial culture system (bioMérieux, Durham, NC), and Lowenstein-Jensen (LJ) solid media by first defining and then investigating contamination suspect positive cultures.
Patients, materials, and methods
Sample collectionAfter written informed consent, 1923 patients undergoing investigation for TB at health centers in Lima, Peru, were recruited over an 18-month period into an operational evaluation of the MODS assay. Two sputum samples were requested from e...