中文摘要：

为建立一种快速检测鸭甲肝病毒1型（DHAV-1）抗体的化学发光酶联免疫分析方法（CLEIA），本研究从DHAV-1中国流行株中扩增获得病毒结构蛋白VP1编码基因，构建重组原核表达质粒pET-32a-VP1，并进行重组蛋白的诱导表达。将纯化后的重组VP1蛋白作为检测抗原建立了CLEIA检测方法。结果显示，利用建立的CLEIA方法对AIV、AMPV、DPV、ARV、MPV、RAV等病毒血清进行检测，均无交叉反应，特异性强；灵敏度高于ELISA方法；重复性试验变异系数为2.18 %～4.33 %，具有较好的重复性；且与中和试验的阳性符合率和阴性符合率分别为100 %和80 %。本研究建立的CLEIA方法可以用于DHAV-1感染监测及疫苗接种后的免疫评价。

英文摘要：

A novel chemiluminescent enzyme immunoassay （CLEIA） was developed to detect antibodies against duck hepatitis A virus type 1（DHAV-1）. The VP1 gene fragment of DHAV-1 was amplified by RT-PCR using DHAV-1 RNA as the template. Recombinant expression plasmid pET-32a-VP1 was constructed, and the induced recombinant protein VP1 （rVP1） was purified using Nickel affinity chromatography. The CLEIA method was established using the purified rVP1 as testing antigen. Meanwhile, its specificity, sensitivity, repeatability and coincidence rate with the neutralization test were evaluated, the results showed that the rVP1 had no crossing reaction with the antibodies to AIV, AMPV, DPV, ARV, MPV, RAV, and the coefficient of variation of the repeatability experiment was 2.18% to 4.33%, indicating that the CLEIA method had high specificity and repeatability. Moreover, CLEIA was more sensitive than ELISA method. Compared with the serum neutralization test, the positive and negative coincidence rate was calculated as 100% and 80%, respectively. The CLEIA method could be used for infection monitoring and immune evaluation after vaccination with DHAV-1.