实时PCR法在慢性前列腺炎病原学诊断中的应用研究

Application of real-time PCR in the pathogenic diagnosis of chronic prostatitis

目的为研究前列腺细菌感染和慢性前列腺炎之间潜在的关联,本文报告了应用实时PCR方法检测前列腺液(EPS)或按摩前列腺后的尿液(VB3)中的细菌。方法细菌模板DNA取自慢性前列腺炎患者的EPS或VB_3标本。1对通用引物:用于扩增所有已知细菌的核糖体DNA(16SrDNA)。3个荧光探针:一个为检测所有细菌的通用探针(Uniprobe);1个为检测大肠杆菌的特异性探针(Ecprobe);1个为检测金黄色葡萄球菌的特异性探针(SAprobe)。结果应用实时PCR方法检测慢性前列腺炎患者,80个标本中有28个检测到细菌,包括14个为大肠杆菌阳性,6个为金黄色葡萄球菌阳性。结论实时PCR具有能定量、快速、特异等优点,有望成为能对慢性前列腺炎患者EPS或VB_3中的细菌进行定性、定量检测的一种很有前途的方法。

Objective To investigate the potential relationship between prostatic bacterial infection and chronic prostatitis （CP）, bacteria in expressed prostate secretions（EPS） or voided bladder 3 （VB3） was detected using real- time PCR. Methods Template DNAs were extracted from EPS or VB3 of patients with chronic prostatitis. A pair of PCR primers were designed for all known bacterial ribosomal DNAs（16SrDNAs）. Three fluorescent probes includ- ing Uniprobe（ all known bacteria）, ECprobe（Escherichia coli） and Saprobe（Staphylococcus aureus） were used in real- time PCR assay. Results Bacteria DNA in 28 samples from patients with chronic prostatitis were found by real-time PCR, including 14 positive samples of E.coli and 6 positive samples of S.aurs. Conclusion Real-time PCR with advantages in quantitative analysis, speed of detection and specificity showed a promising future for the detection of EPS （VB3） bacteria in chronic prostatitis patients.