Colorthink Pro 3 0 3 Serial Dilutions Are Used To

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Quantitative cultures of bronchoalveolar lavage (BAL) fluid are important in the diagnosis of ventilator-associated pneumonia, and calibrated loops are commonly used to set up these cultures. In this study, the performances of calibrated 0.010- and 0.001-ml loops in the transfer of BAL fluid were determined. Five loops of one lot from seven manufacturers were tested. Calibrations were performed by the gravimetric method (0.010-ml loops) and the colorimetric method (0.001-ml loops). Most of the 0.010-ml loops displayed a precision that was less than 10%, but six of them showed very poor accuracies as they transferred a deficiency (nichrome loops) or an excess (disposable loops) of BAL fluid that exceeded ±10%. The mean maximum and minimum BAL fluid volumes delivered by the 0.010-ml loops differed by a factor 3. The 0.001-ml loops displayed acceptable precision.

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Five of them showed inaccuracies of ≤±10%, and mean maximum and minimum BAL fluid volumes had a range of a factor of 2. For all loops, the volumes of BAL fluid sampled were larger than the volumes of reagent-grade water sampled. Results of the colony counting experiments confirmed these findings and revealed a high intra-assay variability for the 0.001-ml loops. We conclude that, when BAL fluid samples are cultured with calibrated loops, (i) proper verification of the calibration of these loops is mandatory, (ii) calibrations should be performed with BAL fluid as the test solution, and (iii) borderline quantitative culture results should be interpreted with knowledge of the inaccuracy values of these loops. Quantitative cultures of bronchoalveolar lavage (BAL) fluid are used in the diagnosis of ventilator-associated pneumonia (VAP). As the dilution of the lung secretions in the BAL fluid is 10- to 100-fold, a colony count of ≥10 4 CFU/ml represents a bacterial load of 10 5 to 10 6/ml in the collection site, which is indicative of bacterial pneumonia (, ).

Conversely, a BAL fluid colony count below the 10 4-CFU/ml threshold points to oropharyngeal contamination. This theoretical concept has been validated in numerous clinical studies, and quantitative culture of BAL fluid specimens is consequently recommended as the reference method for the diagnosis of VAP (). For quantitative cultures of BAL fluid, two approaches are used: the serial dilution method and the calibrated loop method ().

In the serial dilution method, 0.100-ml aliquots of the raw BAL fluid and two serial 100-fold dilutions are inoculated onto the agar plate surfaces. After incubation, counts are made from the dilution that contains the greatest number of bacteria without confluence or overcrowding. Many microbiological laboratories perceive this method as too cumbersome and labor-intensive (). Therefore, they prefer the calibrated loop method, which they are familiar with. Calibrated loops are routinely used to set up quantitative urine cultures (, ).