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首页> 美国卫生研究院文献>Journal of Clinical Microbiology >Highly Reproducible Bactericidal Activity Test Results by Using a Modified National Committee for Clinical Laboratory Standards Broth Macrodilution Technique
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Highly Reproducible Bactericidal Activity Test Results by Using a Modified National Committee for Clinical Laboratory Standards Broth Macrodilution Technique

机译：通过使用改良的全国临床实验室标准肉汤大量稀释技术委员会可高度再现杀菌活性测试结果

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摘要

Bactericidal testing historically has exhibited variable reproducibility, even when prior standardized methods were employed. Several modifications to the National Committee for Clinical Laboratory Standards (NCCLS) broth macrodilution method are proposed to improve reproducibility. Recommended changes from the approved NCCLS guidelines (M21-A and M26-A) include omitting serum supplementation of Mueller-Hinton broth, incubating tubes at 35°C for 24 h with no agitation until they are sampled, running all tests in duplicate with six dilutions instead of nine, reincubating the test for an additional 24 h to resolve discrepant bactericidal activity test results, using a single 0.1-ml sample from each clear tube for subculture, and adopting an alternate method for calculating endpoint determination. In order to test these recommendations in a clinical laboratory setting, we used the modified methodology on 224 separate tests for bactericidal activity. There were 102 serum bactericidal titer (SBT) and 122 minimum bactericidal concentration (MBC) assays performed. By defining reproducibility as agreement between duplicate tests ± 1 dilution, we found 207 of 224 tests (92%) were reproducible at the 24-h subculture point (94% for the SBT assay and 91% for the MBC assay). When the 17 assays with discrepant results were incubated an additional 24 h for a second subculture, only 1 of 224 tests (0.4%) remained discrepant. The method used is practical for a clinical laboratory that chooses to perform bactericidal activity testing and assures a high level of reproducibility between duplicate assays. The total cost of a test was approximately $25.00.
机译：历史上，即使采用先前的标准化方法，杀菌测试也表现出可变的可重复性。提议对美国国家临床实验室标准委员会（NCCLS）肉汤大量稀释方法进行一些修改，以提高可重复性。对已批准的NCCLS指南（M21-A和M26-A）的建议更改包括：省略Mueller-Hinton肉汤的血清补充；在不搅拌的情况下将试管在35°C下孵育24小时，直到对其进行采样；所有测试一式六份进行稀释而不是9种稀释液，再将试验再温育24小时，以解决不同的杀菌活性试验结果，使用每个透明管中的单个0.1 ml样品进行传代培养，并采用另一种方法计算终点测定值。为了在临床实验室环境中测试这些建议，我们对224种单独的杀菌活性测试使用了改进的方法。进行了102次血清杀菌滴度（SBT）和122次最低杀菌浓度（MBC）分析。通过将重复性定义为重复测试±1稀释之间的一致性，我们发现224个测试中的207个（92％）在24小时传代培养点可重复（SBT分析为94％，MBC分析为91％）。当将17个测定结果不一致的样品再培养24小时以进行第二次传代培养时，224个试验中只有1个（0.4％）仍保持差异。所使用的方法对于选择进行杀菌活性测试并确保重复测定之间具有高水平可重复性的临床实验室是实用的。一项测试的总费用约为25美元。

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