Multicenter Comparison Trial of DNA Extraction Methods and PCR Assays for Detection of Chlamydia pneumoniae in Endarterectomy Specimens

The reported rate of detection of Chlamydia pneumoniae DNA within atherosclerotic lesions by PCR varies between 0 and 100%. In this study, identical sets of coded experimental atheroma samples (n = 15) and spiked controls (n = 5) were analyzed by 16 test methods in nine centers by means of PCR. The positive controls were correctly identified to levels of 1, 0.1, and 0.01 inclusion bodies of C. pneumoniae/ml of tissue homogenate by 16 (100%), 11 (69%), and 3 (19%) of the test methods, respectively. Three out of 16 negative controls (19%) were rated positive. Positivity rates for atheroma samples varied between 0 and 60% for the different test methods, with the maximum concordant result for positivity being only 25% for one carotid artery sample. There was no consistent pattern of positive results among the various laboratories, and there was no correlation between the detection rates and the sensitivity of the assay used.

现有研究报道的、通过聚合酶链式反应（PCR）检测动脉粥样硬化病灶内肺炎衣原体（Chlamydia pneumoniae）DNA的阳性检出率区间为0%至100%。本研究中，9家中心的16种检测方法采用PCR对统一编码的15份动脉粥样硬化实验样本（n=15）与5份加标对照样本（n=5）开展了平行分析。针对每毫升组织匀浆中含1、0.1及0.01份肺炎衣原体包涵体的阳性对照样本，分别有16种（100%）、11种（69%）及3种（19%）检测方法可正确识别其浓度水平。16份阴性对照样本中有3份（19%）被误判为阳性。不同检测方法对动脉粥样硬化样本的阳性检出率区间为0%至60%；其中某一份颈动脉样本的阳性检测结果一致性最高，也仅为25%。各实验室间的阳性结果未呈现统一规律，且检测检出率与所用实验方法的灵敏度之间无相关性。