Development of Lateral-flow Immunoassay for WSSV with Polyclonal Antibodies Raised against Recombinant VP (19+28) Fusion Protein

Qing-yu CHENG; Xiao-lin MENG; Jin-ping XU; Wei LU; Jian WANG

February 2007

Qing-yu CHENG, Xiao-lin MENG, Jin-ping XU, Wei LU and Jian WANG. Development of Lateral-flow Immunoassay for WSSV with Polyclonal Antibodies Raised against Recombinant VP (19+28) Fusion Protein[J]. Virologica Sinica, 2007, 22(1): 61-67.

Citation: Qing-yu CHENG, Xiao-lin MENG, Jin-ping XU, Wei LU, Jian WANG. Development of Lateral-flow Immunoassay for WSSV with Polyclonal Antibodies Raised against Recombinant VP (19+28) Fusion Protein .VIROLOGICA SINICA, 2007, 22(1) : 61-67.

应用胶体金免疫层析法快速检测对虾白斑综合症病毒（WSSV）

武汉大学生命科学学院病毒学国家重点实验室，湖北武汉 430072

通讯作者： 孟小林*, mengxiaolin8@hotmail.com;
收稿日期： 2006-06-20
录用日期： 2006-07-18

摘要

摘要：利用免疫胶体金结合免疫层析法研制了一种快速检测对虾白斑综合症病毒（WSSV）的试剂条。利用大肠杆菌表达系统融合表达了WSSV囊膜蛋白VP (19+28)，然后用纯化后的融合蛋白VP (19+28) 作为抗原免疫家兔，制备和纯化了抗WSSV- VP (19+28) IgG。以胶体金标记此抗体制成免疫层析试剂条。此试剂条可以从体液、腮、肝胰腺三种不同组织中检测出WSSV的感染；而且从垂死虾中也可以检测出WSSV的感染；以纯化的VP (19+28)为样品，试剂条的检测下限可达到10ng/mL。同时用WSSV vp 19 的引物进行了PCR检测。二种检测方法得到了完全一致的结果。
- 对虾白斑综合症病毒（WSSV）
- , VP19
- , VP28
- , 胶体金

Development of Lateral-flow Immunoassay for WSSV with Polyclonal Antibodies Raised against Recombinant VP (19+28) Fusion Protein

State Key Laboratory of Virology,College of Life Science,Wuhan University,Wuhan,430072,People's Republic of China

Corresponding author: Xiao-lin MENG, mengxiaolin8@hotmail.com
Received Date: 20 June 2006
Accepted Date: 18 July 2006

Abstract

We developed a sensitive and rapid lateral-flow immunoassay (LFIA) for WSSV, using a colloidal gold as an indicator. The fusion protein, VP (19+28), was expressed in E. coli, purified and used to prepare polyclonal antibodies. The purified anti-VP (19+28) IgG were conjugated with colloidal gold. Unconjugated anti-VP (19+28) IgG and goat anti-rabbit IgG were immobilized on nitrocellulose membranes. After assembly, three groups (5 individual animals in each group)of shrimp samples were tested which included healthy, moribund and dead shrimps. For each group, three different tissues (body juices, gills and hepatopancreas) were tested at the same time. In parallel, all the samples were also analyzed using an existing PCR for comparison. Out of 45 samples tested, 30 were detected as positive while 15 negative The results of LFIA correlate with those obtained by the PCR analysis, indicating that these two detection methods have the same efficacy in the limited number of samples tested in this preliminary study.
- Lateral-flow immunoassay
- , White spot syndrome virus (WSSV)
- , VP19
- , VP28

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