中文摘要：

目的兔戊型肝炎病毒（HEV）是近年来新发现的,根据其分子生物学特征可能成为HEV的新基因型。为提高对兔HEV的检出率,本研究设计了针对兔HEV RNA的特异性引物和TaqMan探针并对其进行考评。方法 1）从GenBank中下载14条兔HEV全长序列,分别在其ORF1、ORF3片段的保守区域设计一条TaqMan探针和一对上、下游特异性引物,其中ORF1片段的探针及引物为C组,ORF3片段的为B组;A组的探针及引物为Jothikumar N等所报道的通用引物;2）制备相应的质粒标准品来构建定量标准曲线,并利用上述3套引物、探针对49份兔粪便及44份兔血液样品进行检测,将B、C组检测结果与A组的检测结果进行比较。结果新建立了两套兔HEV定量PCR的TaqMan探针及引物（B、C组）,其标准曲线的斜率分别为-3.455和-3.469;使用上述3套引物及探针（A、B、C组）对49份粪便标本进行荧光定量PCR检测,其HEV RNA阳性率分别为67.35%（33/49）、67.35%（33/49）、57.14%（28/49）,平均HEV RNA（log copies/g）拷贝数为6.18、5.90、6.11;44份血液标本的HEV RNA阳性检出率分别为65.90%（29/44）、56.82%（25/44）、50.00%（22/44）,平均HEV RNA（log copies/mL）拷贝数为3.62、3.43、3.03;结果显示以上3组探针及引物均可用于粪便和血液标本的定量检测。结论兔HEV虽然有其独特的基因结构,但使用HEV通用引物检测不影响检出率。

英文摘要：

In order to improve the detection rate for rabbit HEV RNA, specific primers and TaqMan probes were designed and evaluated in this study. Fourteen complete genome sequences of rabbit HEV isolates downloaded from GenBank were aligned to get the conserved areas in ORF1 and ORF3. Subsequently, TaqMan probes and pairs of primers were designed and synthesized according to the conserved areas respectively. The probe and primers for ORF1 were named as group C and ORF3 as group B, while those had been reported by Jothikumar N were named as group A. Then three groups of different serial 10-fold dilutions of standard plasmid stocks were constructed. The standard curves were generated, and then 49 feces and 44 sera collected from rabbits were tested by the three groups of probes and primers. The results were compared and analyzed. Two newly-designed TaqMan probes and primers for rabbit HEV detection were established and the slopes of the standard curves were -3. 455 and -3. 469. The positive rates of the three groups （A, B and C） for detection of the 49 rabbit feces were 67.35% （33/49）, 67. 350% （33/49）, 57.14% （28/49） and the mean viral load of HEV RNA （log copies/g） was 6.18, 5.90 and 6.11, respectively. In addition, the positive rates of serum HEV RNA were 65. 90% （29/44）, 56. 82% （25/44） and 50.00% （22/44） and the mean viral load of HEV RNA （log copies/mL） was 3.62, 3.43 and 3.03. Though varied in genomic structure, the application of universal TaqMan probes and primers for rabbit HEV detection does not affect its results.