An improved indirect ELISA for specific detection of antibodies against classical swine fever virus based on structurally designed E2 protein expressed in suspension mammalian cells
文献类型: 外文期刊
第一作者: Ji, Shengwei
作者: Ji, Shengwei;Luo, Yuzi;Zhang, Tengteng;Shao, Lina;Meng, Xing-Yu;Wang, Yimin;Gao, Yao;Li, Yongfeng;Li, Su;Sun, Yuan;Qiu, Hua-Ji;Ji, Shengwei;Zhang, Tengteng;Jin, Xin
作者机构:
期刊名称:ARCHIVES OF VIROLOGY ( 影响因子:2.574; 五年影响因子:2.466 )
ISSN: 0304-8608
年卷期: 2018 年 163 卷 7 期
页码:
收录情况: SCI
摘要: Classical swine fever (CSF), which is caused by classical swine fever virus (CSFV), is a highly contagious disease of pigs. CSFV is genetically and serologically related to bovine viral diarrhea virus (BVDV), a ruminant pestivirus. However, currently available ELISAs based on the full-length E2 protein of CSFV cannot discriminate anti-CSFV from anti-BVDV antibodies. In this study, a truncated CSFV E2 protein (amino acids 690 to 879) covering antigenic domains B/C/D/A (E2(B/C/D/A)) was designed based on homologous modeling according to the crystal structure of the BVDV E2 protein. The E2(B/C/D/A) protein was expressed in CHO cells adapted to serum-free suspension culture, and an indirect ELISA (iELISA) was established based on the recombinant protein. No serological cross-reaction was observed for anti-BVDV sera in the iELISA. When testing 282 swine serum samples, the iELISA displayed a high sensitivity (119/127, 93.7%) and specificity (143/155, 92.3%), with an agreement of 92.9% (262/282) and 92.2% (260/282) with virus neutralization test and the IDEXX CSFV blocking ELISA, respectively. Taken together, the newly developed iELISA is highly specific and sensitive and able to differentiate anti-CSFV from anti-BVDV antibodies.
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