鸭甲型肝炎病毒1型抗体CLEIA检测方法的建立

发布时间：2018-02-21 09:30

本文关键词： 鸭病毒性肝炎 VP 化学发光酶联免疫　出处：《中国预防兽医学报》2017年06期 　论文类型：期刊论文

【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。
[Abstract]:In order to establish a chemiluminescent enzyme linked immunosorbent assay for rapid detection of duck hepatitis A virus type 1 (DHAV-1) antibody, the VP1 encoding gene of virus structural protein was obtained from a Chinese epidemic strain of DHAV-1. The recombinant prokaryotic expression plasmid pET-32a-VP1 was constructed, and the recombinant protein was induced to express. The purified recombinant VP1 protein was used as the detection antigen to establish a CLEIA detection method. There was no cross reaction, high specificity, sensitivity higher than ELISA method, the coefficient of variation of reproducibility test was 2.18 and 4.33, which had good repeatability. The positive coincidence rate and negative coincidence rate with neutralization test were 100% and 80 respectively. The CLEIA method established in this study can be used for DHAV-1 infection monitoring and immunization evaluation after vaccination.
【作者单位】：广西大学动物科学技术学院;中国科学院微生物研究所病原微生物与免疫学重点实验室;中国兽医药品监察所;
【基金】：国家重点研发计划“畜禽重大疫病防控与高效安全养殖综合技术研发”重点专项(2016YFD0500800) 中国科学院重点部署项目(ZDRW-ZS-2016-4-4、KSZD-EW-Z-005-001)
【分类号】：S852.65

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