定量检测HCMV pp67 mRNA快速诊断人巨细胞病毒活动性感染  

The quantitative detection of human cytomegalovirus pp67 mRNA in rapid diagnosis of active human cytomegalovirus infection

作　　者：钟峰[1] 赵俊[1] 张业婷 姚瑶[1] 王明丽[1] 

机构地区：[1]安徽医科大学微生物学教研室,合肥230032

出　　处：《安徽医科大学学报》2016年第3期324-328,共5页Acta Universitatis Medicinalis Anhui

基　　金：国家自然科学基金(编号:30872253);安徽省教育厅自然科学重大研究项目(编号:KJ2012ZD08)

摘　　要：目的 建立实时荧光定量逆转录PCR（Real time RT-qPCR）方法,检测外周血中人巨细胞病毒（HCMV）pp67mR-NA的拷贝数,用于临床快速诊断HCMV活动性感染。方法采用RT-qPCR测定86份癌症患者外周血标本中的HC-MVpp67mRNA拷贝数,并与HCMVpp65抗原血症检测及HCMV分离培养的实验结果进行比较。结果 86例癌症患者中,HCMV病毒分离检测阳性率3.49%（3例）;实时荧光定量RT-PCR检测阳性率17.44%（15例）,灵敏度为100%,特异性为85.54%;HCMV pp65抗原血症检测阳性率为10.47%（9例）,灵敏度为66.7%,特异度为92.8%。两者比较差异有统计学意义（P〈0.05）。通过受试者工作特征（ROC）曲线分析,RT-qPCR检测每2×105个外周血白细胞中HCMV拷贝数的截断点值为201.5;此病毒拷贝数可作为诊断HCMV活动性感染的参考阈值。结论 应用RT-qPCR检测HCMV活动性感染,灵敏度高、特异性好;并能够为抗病毒治疗供重要的参考依据。Objective To explore the detection of HCMV pp67 mRNA in peripheral blood by RT-PCR for the diagnosis of HCMV active infection. Methods The peripheral blood samples of 86 postoperative cancer patients were measured by the real-time quantitative PCR（RT-PCR）. The results were compared with pp65 antigenemia test and HCMV virus isolation. Results In the 86 cancer patients, 15 patients were positive in pp67 mRNA RT-PCR, 9 pa- tients were positive in pp65 antigenemia test, 3 patients were positive in virus isolation. The sensitivity of HCMV pp67 mRNA RT-PCR was 100%, and the specificity was 87.9%, the sensitivity of HCMV pp65 antigenemia was 66. 7% and the specificity was 92. 8% , the difference was statistically significant（ P 〈 0. 05 ）. The result of ROC curve analysis showed the cutoff value of real-time RT-qPCR is 201.5copies/2 x 105 Peripheral blood leucocyte cells, these hint this virus content can be used as a reference threshold of HCMV active infection. Conclusion The sensitivity and specificity of the RT-qPCR are good,which can provide fast and accurate diagnosis of HCMV active infection, and lay the basis for clinical antiviral treatment.

关 键 词：人巨细胞病毒 pp67mRNA 实时荧光定量RT-PCR 活动性感染 

分 类 号：R373.9[医药卫生—病原生物学]