DOI number:10.16303/j.cnki.1005-4545.2021.07.02
Affiliation of Author(s):Yangzhou University
Journal:Chinese journal of veterinary science
Key Words:Porcine Deltacoronavirus, Nucleocapsid(N) protein, Prokaryotic expression, Indirect ELISA
Abstract:Ｔｈｅ ａｍｉｎｏ ａｃｉｄ ｓｅｑｕｅｎｃｅ ｏｆ ｐｏｒｃｉｎｅ Ｄｅｌｔａｃｏｒｏｎａｖｉｒｕｓ（ＰＤＣｏＶ）Ｎ ｐｒｏｔｅｉｎ ｗａｓ ａｎａｌｙｚｅｄｂｙ ＤＮＡＳｔａｒ Ｐｒｏｔｅａｎ ｓｏｆｔｗａｒｅ，ａｎｄ ｔｈｅ ｒｅｇｉｏｎ ｗｉｔｈ ｓｔｒｏｎｇ ａｎｔｉｇｅｎｉｃｉｔｙ（２６５－３４２ａａ）ｗａｓ ｓｅｌｅｃｔｅｄ．Ａｃｃｏｒｄｉｎｇ ｔｏ ｔｈｅ ｆｒｅｑｕｅｎｃｙ ｏｆ Ｅ．ｃｏｌｉ ｃｏｄｏｎｓ，ｔｈｅ ｏｐｔｉｍｉｚｅｄ ｄｅｓｉｇｎ ｗａｓ ｃａｒｒｉｅｄ ｏｕｔ ａｎｄ ｔｈｅ ｃｏｒｒｅｓｐｏｎｄｉｎｇ ｇｅｎｅ ｆｒａｇｍｅｎｔ Ｎ－ｏｐｔｉ ｗａｓ ａｒｔｉｆｉｃｉａｌｌｙ ｓｙｎｔｈｅｓｉｚｅｄ ａｎｄ ｃｌｏｎｅｄ ｉｎｔｏ ｐＥＴ３２ａｆｏｒ ｔｈｅ ｐｒｏｋａｒｙｏｔｉｃ ｅｘｐｒｅｓｓｉｏｎ ｏｆ Ｎ ｐｒｏｔｅｉｎ．Ｔｈｅ ｏｂｔａｉｎｅｄ ｆｕｓｉｏｎ ｐｒｏｔｅｉｎ ｗａｓ ｌａｂｅｌｅｄ ｗｉｔｈ Ｈｉｓ－ｔａｇ ａｎｄ ｎａｍｅｄａｓ ＰＤＣｏＶ－Ｎ－Ｈｉｓ，ｉｔ ｗａｓ ａ ｓｏｌｕｂｌｅ ｐｒｏｔｅｉｎ．Ｔｈｅ Ｗｅｓｔｅｒｎ ｂｌｏｔ ｒｅｓｕｌｔｓ ｓｈｏｗｅｄ ｔｈａｔ ｔｈｅ ｓｉｚｅ ｏｆ ｔｈｅ ｒｅｃｏｍｂｉｎａｎｔ Ｎ ｐｒｏｔｅｉｎ ｗａｓ ａｂｏｕｔ ３１ｋＤａ．Ｔｈｅ ｒｅｃｏｍｂｉｎａｎｔ Ｎ ｐｒｏｔｅｉｎ ｗａｓ ｐｕｒｉｆｉｅｄ ｂｙ Ｎｉ－ＮＴＡ ａｆｆｉｎｉｔｙ ｃｈｒｏｍａｔｏｇｒａｐｈｙ ａｎｄ ｕｓｅｄ ａｓ ｔｈｅ ｃｏａｔｉｎｇ ａｎｔｉｇｅｎ．Ｂｙ ｏｐｔｉｍｉｚｉｎｇ ｔｈｅ ｒｅａｃｔｉｏｎ ｃｏｎｄｉｔｉｏｎｓ，ａｎ ｉｎｄｉｒｅｃｔ ＥＬＩＳＡ ｍｅｔｈｏｄ ｆｏｒ ｄｅｔｅｃｔｉｏｎ ｏｆ ＰＤＣｏＶ ｓｐｅｃｉｆｉｃ ａｎｔｉｂｏｄｙ ｗａｓ ｅｓｔａｂｌｉｓｈｅｄ．Ｔｈｅ ｒｅｓｕｌｔｓ ｓｈｏｗｅｄ ｔｈａｔ ｔｈｉｓ ｉｎｄｉｒｅｃｔ ＥＬＩＳＡ ｈａｄ ｇｏｏｄ ｓｐｅｃｉｆｉｃｉｔｙ ａｎｄ ｒｅｐｅａｔａｂｉｌｉｔｙ，ｓｉｎｃｅ ｉｔ ｈａｓ ｎｏ ｃｒｏｓｓ－ｒｅａｃｔｉｏｎ ｗｉｔｈ ｔｈｅ ｐｏｓｉｔｉｖｅ ｓｅｒｕｍ ｏｆ ＰＥＤＶ，ＴＧＥＶ ａｎｄ ＰＲｏＶ，ｔｈｅ ｖａｒｉａｔｉｏｎ ｃｏｅｆｆｉｃｉｅｎｔ ｏｆ ｔｈｅ ｉｎｔｅｒ－ｂａｔｃｈ ａｎｄ ｉｎｔｒａ－ｂａｔｃｈ ｒｅｐｅａｔａｂｉｌｉｔｙ ｔｅｓｔ ｗａｓ ｌｅｓｓ ｔｈａｎ １０％．Ｔｈｅ ｉｎｄｉｒｅｃｔ ＥＬＩＳＡ ｗａｓ ｕｓｅｄ ｔｏ ｄｅｔｅｃｔ ２０ＰＤＣｏＶ ｐｏｓｉｔｉｖｅ ｓｅｒｕｍ ａｎｄ ２０ｎｅｇａｔｉｖｅ ｓｅｒｕｍ，ａｎｄ ｔｈｅ ｒｅｓｕｌｔｓ ｓｈｏｗｅｄ ｔｈａｔ ｔｈｅ ｐｏｓｉｔｉｖｅ ａｎｄ ｎｅｇａｔｉｖｅ ｃｏｉｎｃｉｄｅｎｃｅ ｒａｔｅ ｗａｓ ｂｏｔｈ １００％．Ｔｈｅ ｒｅｓｕｌｔｓ ｓｈｏｗｅｄ ｔｈａｔ ｔｈｅ ｉｎｄｉｒｅｃｔ ＥＬＩＳＡ ｆｏｒ ＰＤＣｏＶ ａｎｔｉｂｏｄｉｅｓ ｈａｓ ｇｏｏｄ ｓｐｅｃｉｆｉｃｉｔｙ ａｎｄ ｃａｎ ｂｅ ｕｓｅｄ ｆｏｒ ｒａｐｉｄ ｄｉａｇｎｏｓｉｓ ａｎｄ ｅｐｉｄｅｍｉｏｌｏｇｉｃａｌ ｉｎｖｅｓｔｉｇａｔｉｏｎ ｏｆ ＰＤＣｏＶ ｉｎｆｅｃｔｉｏｎ．
Co-author:Xueyan Bai,Chengcheng Zhang,Mengjiao Guo,Mengjiao Li,Kai Liao,Feng Xue,Yantao Wu
First Author:Bingxu Qian
Indexed by:Research Atricle
Correspondence Author:Xiaorong Zhang
Document Code:1005-4545（2021）07-1234-08
Discipline:Agricultural science
First-Level Discipline:Veterinary Medicine
Document Type:J
Volume:41
Issue:7
Page Number:1234-1241
Translation or Not:no
Date of Publication:2021-07-15
