Development of a double-antibody sandwich enzyme-linked immunosorbent assay (DAS-ELISA) for the detection of KHV
文献类型： 外文期刊
作者： Li, Yingying 1 ; Wang, Qing 1 ; Hu, Feng 1 ; Wang, Yingying 1 ; Bergmann, Sven M. 3 ; Zeng, Weiwei 4 ; Yin, Jiyuan 1 ; Sh 1 ;
作者机构： 1.Chinese Acad Fishery Sci, Pearl River Fisheries Res Inst, Key Lab Aquat Anim Immune Technol Guangdong Prov, Key Lab Fishery Drug Dev,Minist Agr, Guangzhou, Peoples R China
2.Shanghai Ocean Univ, Coll Fisheries & Life Sci, Shanghai, Peoples R China
3.Fed Res Inst Anim Hlth, Friedrich Loffler Inst FLI, Inst Infectol, German Reference Lab KHVD, Greifswald, Germany
4.Foshan Univ, Sch Life Sci & Engn, Guangdong Prov Key Lab Anim Mol Design & Precise, Foshan, Peoples R China
关键词： double‐ antibody sandwich ELISA (DAS‐ ELISA); Koi herpesvirus (KHV); ORF132; polyclonal antibody
期刊名称：JOURNAL OF FISH DISEASES （ 影响因子：2.767； 五年影响因子：2.689 ）
ISSN： 0140-7775
年卷期：
页码：
收录情况： SCI
摘要： Koi herpesvirus disease (KHVD) caused by the koi herpesvirus (KHV) is difficult to diagnose in live fish, presenting a challenge to the koi industry. The enzyme-linked immunosorbent assay (ELISA) method cannot be widely used to detect KHV because few commercial anti-KHV antibody exists. Here, we developed an anti-ORF132 polyclonal antibody and confirmed its reactivity via indirect immunofluorescence assay and Western blotting. A double-antibody sandwich ELISA (DAS-ELISA) was established to detect KHV, monoclonal antibody 1B71B4 against ORF92 was used as the capture antibody, and the detection antibody was the polyclonal antibody against the truncated ORF132. The lowest limit was 1.56 ng/ml KHV. Furthermore, the DAS-ELISA reacted with KHV isolates, while no cross-reactions occurred with carp oedema virus, spring viraemia of carp virus, frog virus 3 and grass carp reovirus. Two hundred koi serum samples from Guangdong, China, were used in the DAS-ELISA test, and the positive rate of the koi sera was 13%. The clinical sensitivity and specificity of the DAS-ELISA relative to the traditional PCR method were 66.7% and 97.6%, respectively. Our findings may be useful for diagnosing and preventing KHVD in koi and common carp.
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