戊型肝炎患者粪便中病毒RNA的半定量检测

Semi-quantitative detection of hepatitis E virus RNA from stools of patients with hepatitis E

目的 探讨戊型肝炎病毒 (HEV)RNA半定量检测的意义和可行性 ,为HEVRNA的定量PCR(多聚酶链反应 )检测提供科学依据。方法 在逆转录 -套式PCR基础上 ,采用样品倍比稀释法对戊型肝炎患者粪便中病毒RNA作半定量检测。结果 2 0份粪便标本中有 14份检测结果阳性 ,其中病毒RNA拷贝水平为 8× 10 5、8× 10 7、8× 10 8、8× 10 9PCR单位·ml-1的分别为 2、10、1、1份。对同一标本而言 ,在不同稀释度时 ,其PCR产物在量上无显著差异。结论 定量PCR技术检测戊型肝炎病毒RNA是可行的 。

Objective To investigate the significance and feasibility of quantification of hepatitis E virus RNA from stools of patients with hepatitis E and give scientific proof for quantitative detection of it by RT-nested PCR and detection of RNA of viruses.Methods A semi-quantitative detection of HEV RNA was made by serial dilution of the specimens on the basis of RT-nested PCR.Results There were 14 specimens positive among the 20 clinical specimens and the copies of RNA were about 8×10 5 PCR unit·ml -1 for 2 specimens,8×10 7PCR unit·ml -1 for 10 specimens,8×10 8 PCR unit·ml -1 and 8×10 9PCR unit·ml -1 each for 1 specimen.But there was little difference among the amount of final products for HEV RNA RT-nested PCR.Conclusion The quantification of HEV RNA by RT-nested PCR is feasible but the quantity of HEV RNA should be determined by the initial cDNA not PCR products.