套式PCR法检测外周血肺炎衣原体DNA

目的探讨外周血单个核细胞(PBMC)中肺炎衣原体(Cpn)DNA的检测方法.方法用淋巴细胞分层法和低渗溶血法提取PBMC.设计2对引物用套式PCR(nPCR)法测定150例冠心病患者(冠心病组)外周血PBMC中Cpn-DNA.同期临床55例非冠心病患者作对照组.结果冠心病患者肺炎衣原体DNA阳性率32.7%,对照组阳性率为1.8%,两组差异显著(P＜0.001),OR 26.2,95%CI 3.52～194.98.冠心病组中无症状组阳性率为29.1%,不稳定型心绞痛组阳性率为39.6%,急性心肌梗死组阳性率为29.9%,各亚组间差异不显著(P＞0.05).结论用nPCR法检测PBMC中Cpn-DNA,可为临床Cpn感染的诊断提供依据.

Detection of Chlamydia pneumoniae DNA in peripheral blood mononuclear cell by nested polymerase chain reaction

Objective To investigate the assay for detection of chlamydia pneumoniae (cpn) DNA in the peripheral blood mononuclear cell using nested polymerase chain reaction (nPCR).Methods Peripheral blood mononuclear cells(PBMC) were collected by using lymphocyte Ficoll and the low-osmotic hemolysis method.Cpn DNA was examined with nPCR.All 150 patients with coronary heart disease (CHD) were confirmed by angiography,and another 55 clinical CHD patients with normal angiography were characterized as control.Results Chlamydia pneumoniae DNA was detected in 49 (32.7%) of 150 patients with CHD and in 1 (1.8%) of 55 of control group.The positive rate of nPCR in CHD groups was significantly higher than those in control group.OR was 26.2 and 95%CI (confidence interval) was 3.52-194.98.Conclusion PBMC sample could be collected easily.The nPCR is sensitive and specific method,and could be used for clinical diagnosis of Cpn infection in coronary heart disease patients.