建立用于检测水貂冠状病毒抗体的血清学检测方法。

Establishment of serological test to detect antibody against ferret coronavirus.

作者信息

Minami Shohei, Terada Yutaka, Shimoda Hiroshi, Takizawa Masaki, Onuma Mamoru, Ota Akihiko, Ota Yuichi, Akabane Yoshihito, Tamukai Kenichi, Watanabe Keiichiro, Naganuma Yumiko, Kanagawa Eiichi, Nakamura Kaneichi, Ohashi Masanari, Takami Yoshinori, Miwa Yasutsugu, Tanoue Tomoaki, Ohwaki Masao, Ohta Jouji, Une Yumi, Maeda Ken

机构信息

Laboratory of Veterinary Microbiology, Joint Faculty of Veterinary Medicine, Yamaguchi University, 1677-1 Yoshida, Yamaguchi 753-8515, Japan.

出版信息

J Vet Med Sci. 2016 Jul 1;78(6):1013-7. doi: 10.1292/jvms.16-0059. Epub 2016 Mar 3.

DOI:10.1292/jvms.16-0059

PMID:26935842

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC4937135/

Abstract

Since there is no available serological methods to detect antibodies to ferret coronavirus (FRCoV), an enzyme-linked immunosorbent assay (ELISA) using recombinant partial nucleocapsid (N) proteins of the ferret coronavirus (FRCoV) Yamaguchi-1 strain was developed to establish a serological method for detection of FRCoV infection. Many serum samples collected from ferrets recognized both a.a. 1-179 and a.a. 180-374 of the N protein, but two serum samples did not a.a. 180-374 of the N protein. This different reactivity was also confirmed by immunoblot analysis using the serum from the ferret.Therefore, the a.a. 1-179 of the N protein was used as an ELISA antigen. Serological test was carried out using sera or plasma of ferrets in Japan. Surprisingly, 89% ferrets in Japan had been infected with FRCoV. These results indicated that our established ELISA using a.a. 1-179 of the N protein is useful for detection of antibody to FRCoV for diagnosis and seroepidemiology of FRCoV infection.

摘要

由于没有可用的血清学方法来检测雪貂冠状病毒（FRCoV）抗体，因此开发了一种使用雪貂冠状病毒山口-1株重组部分核衣壳（N）蛋白的酶联免疫吸附测定（ELISA），以建立一种检测FRCoV感染的血清学方法。从雪貂收集的许多血清样本识别N蛋白的第1-179位氨基酸和第180-374位氨基酸，但有两个血清样本不识别N蛋白的第180-374位氨基酸。使用雪貂血清进行的免疫印迹分析也证实了这种不同的反应性。因此，N蛋白的第1-179位氨基酸被用作ELISA抗原。使用日本雪貂的血清或血浆进行了血清学检测。令人惊讶的是，日本89%的雪貂已感染FRCoV。这些结果表明，我们建立的使用N蛋白第1-179位氨基酸的ELISA对于检测FRCoV抗体以用于FRCoV感染的诊断和血清流行病学研究是有用的。