戊型肝炎患者粪便中病毒RNA的半定量检测

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20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8×105、8×107、8×108、8×109PCR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，但应以起始模板来定量而不是以终产物定量。

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

Objective To investigate the significance and feasibility of quantification of hepatitis E virus RNA from stools of pa-tients with hepatitis E and give scientific proof for quantitative detection of it by RT- nested PCR and detection of RNA of viruses. Meth-ods 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 anong the 20 clinical specimens and the copies of RNA were about 8×105 PCR unit·ml- 1 for 2 spec-imens,8×107pCR unit·ml- 1 for 10 specimens,8×108 PCR unit·ml- 1 and 8×109PCR unit·ml- 1 each for 1 specimen. But there was lit-tle 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.