Ciervo Alessandra, Petrucca Andrea, Cassone Antonio
Department of Bacteriology and Medical Mycology, Istituto Superiore di Sanità, Viale Regina Elena 299, 00161 Rome, Italy.
Mol Cell Probes. 2003 Apr-Jun;17(2-3):107-11. doi: 10.1016/s0890-8508(03)00028-8.
A real-time PCR assay for Chlamydia pneumoniae in human atherosclerotic plaques by the use of novel probes and FRET LightCycler technology, is described. The assay proved particularly suitable for the specific and quantitative detection of a low DNA copy number in conventional PCR-negative samples. Among fifteen nested-PCR negative atherosclerotic plaques examined, our method detected three positive plaques containing 50(+/-3), 37(+/-2) and 24(+/-2) DNA copy number+/-SD in three independent experiments. Real-time PCR holds promise for C. pneumoniae quantitation in human atherosclerotic plaques.
描述了一种使用新型探针和荧光共振能量转移(FRET)LightCycler技术对人动脉粥样硬化斑块中的肺炎衣原体进行实时聚合酶链反应(PCR)检测的方法。该检测方法特别适用于在传统PCR阴性样本中特异性和定量检测低DNA拷贝数。在检测的15个巢式PCR阴性的动脉粥样硬化斑块中,我们的方法在三个独立实验中检测到三个阳性斑块,其DNA拷贝数分别为50(±3)、37(±2)和24(±2)。实时PCR在人动脉粥样硬化斑块中肺炎衣原体定量方面具有前景。