Jacobs J A, De Brauwer E I, Cornelissen E I, Drent M
Departments of Medical Microbiology, University Hospital Maastricht, Maastricht, The Netherlands.
J Clin Microbiol. 2000 Jun;38(6):2117-21. doi: 10.1128/JCM.38.6.2117-2121.2000.
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. 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.
支气管肺泡灌洗(BAL)液的定量培养在呼吸机相关性肺炎的诊断中很重要,校准环通常用于进行这些培养。在本研究中,测定了校准的0.010 ml和0.001 ml环在转移BAL液中的性能。测试了来自七个制造商的一批五个环。通过重量法(0.010 ml环)和比色法(0.001 ml环)进行校准。大多数0.010 ml环的精密度小于10%,但其中六个环的准确度很差,因为它们转移的BAL液不足(镍铬合金环)或过量(一次性环)超过了+/-10%。0.010 ml环输送的BAL液平均最大和最小体积相差3倍。0.001 ml环显示出可接受的精密度。其中五个环的误差≤+/-10%,BAL液平均最大和最小体积的范围为2倍。对于所有环,采集的BAL液体积均大于采集的试剂级水体积。菌落计数实验结果证实了这些发现,并揭示了0.001 ml环在实验内的高变异性。我们得出结论,当用校准环培养BAL液样本时,(i)必须对这些环的校准进行适当验证,(ii)应以BAL液作为测试溶液进行校准,(iii)应结合这些环的误差值来解释临界定量培养结果。