Department of Microbiology, University of Antwerp, Antwerp, Belgium.
J Microbiol Methods. 2010 Aug;82(2):131-5. doi: 10.1016/j.mimet.2010.05.006. Epub 2010 May 19.
An external quality assessment (EQA) panel consisting of a total of 13 samples in broncho alveolar lavage (BAL) or transport medium was prepared to assess the proficiency of laboratories in the correct detection of Chlamydophila pneumoniae and Mycoplasma pneumoniae by nucleic acid amplification techniques (NAATs) (6 samples containing various concentrations (4.9-490 inclusion forming units (IFU)/ml) of C. pneumoniae, 5 samples containing various concentrations (20-5000 color-changing units (CCU)/ml) of M. pneumoniae and 2 samples negative for both). Seventy-nine laboratories from 18 countries participated in this EQA study. Sixty-four datasets were returned for C. pneumoniae (n=5 conventional commercial, n=10 conventional in-house, n=4 real-time commercial, n=43 real-time in-house, and n=2 SDA). Sixty-seven datasets were obtained for M. pneumoniae (n=5 conventional commercial, n=10 conventional in-house, n=4 real-time commercial, n=46 real-time in-house, and n=2 strand displacement amplification (SDA)). For the total panels, correct results per sample varied between 95.3% and 100% for C. pneumoniae and between 53.7% and 95.5% for M. pneumoniae. In general, commercial conventional NAATs showed possible sensitivity issues when compared to conventional in-house NAATs for both organisms. On the other hand, real-time commercial NAATs scored better than real-time in-house assays in terms of sensitivity for both organisms. For C. pneumoniae and M. pneumoniae, 0.8% and 2.2% true false-positive results and 1.9% and 2.0% false positives were reported in the samples spiked with the other organism. Analysis of the data for C. pneumoniae showed that the concentrations used were easily detectable by the vast majority of participants. The percentage of correct qualitative results for M. pneumoniae demonstrated that the concentrations included in this panel proved challenging for a number of participants.
一个由总共 13 个支气管肺泡灌洗液(BAL)或运输介质样本组成的外部质量评估(EQA)小组被制备来评估实验室在使用核酸扩增技术(NAAT)正确检测肺炎衣原体和肺炎支原体方面的熟练度(6 个样本包含各种浓度(4.9-490 包涵体形成单位(IFU)/ml)的肺炎衣原体,5 个样本包含各种浓度(20-5000 变色单位(CCU)/ml)的肺炎支原体,以及 2 个样本均为两者阴性)。来自 18 个国家的 79 个实验室参与了这项 EQA 研究。共收到 64 个数据集用于肺炎衣原体(n=5 个常规商业,n=10 个常规内部,n=4 个实时商业,n=43 个实时内部,n=2 个单链位移扩增(SDA))。共收到 67 个数据集用于肺炎支原体(n=5 个常规商业,n=10 个常规内部,n=4 个实时商业,n=46 个实时内部,n=2 个链置换扩增(SDA))。对于总样本,每个样本的正确结果在肺炎衣原体之间为 95.3%至 100%,在肺炎支原体之间为 53.7%至 95.5%。一般来说,与两种病原体的常规内部 NAAT 相比,商业常规 NAAT 显示出可能的灵敏度问题。另一方面,在灵敏度方面,实时商业 NAAT 比实时内部检测要好。对于肺炎衣原体和肺炎支原体,在添加另一种病原体的样本中报告了 0.8%和 2.2%的真阳性错误结果和 1.9%和 2.0%的假阳性结果。对肺炎衣原体数据的分析表明,绝大多数参与者很容易检测到使用的浓度。肺炎支原体的定性结果正确百分比表明,该小组中包含的浓度对许多参与者来说具有挑战性。
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