Development of an ELISA based on a multi-fragment antigen of infectious bronchitis virus for antibodies detection.
OBJECTIVE
To develop a cost-effective ELISA for detection of antibodies against infectious bronchitis virus (IBV) by using a multi-fragment protein as coating antigen.


RESULTS
A multi-fragment antigen, termed BE, which was composed of eight antigenic fragments selected from the three major proteins (S, M, and N) of IBV, was expressed in Escherichia coli. The entire protein had a molecular weight of 61.5 kDa. In addition to it, a smaller truncated protein was also produced; both could react with IBV-positive serum. Next, an indirect ELISA (BE-ELISA) was developed. Coefficients of variation of this assay were lower than 10 %, and no cross-reactivity between the coated antigen BE and antiserum against newcastle disease virus, avian influenza virus, or infectious bursal disease virus was observed. The performance of BE-ELISA was evaluated, and showed 95.4 % coincidence ratio with the whole virus based-ELISA (IDEXX).


CONCLUSIONS
The multi-fragment antigen (BE) may represent a promising alternative to the whole virus without safety problems, and this newly established ELISA provides an effective method for anti-IBV antibody detection.
School of Life Science, Sichuan University, Animal Disease Prevention and Food Safety Key Laboratory of Sichuan Province, "985 Project" Science Innovative Platform for Resource and Environment Protection of Southwestern, Chengdu, 610064, Sichuan Province, China. dingmengdie1987@126.com.
Meng-Die
MD
Ding
School of Life Science, Sichuan University, Animal Disease Prevention and Food Safety Key Laboratory of Sichuan Province, "985 Project" Science Innovative Platform for Resource and Environment Protection of Southwestern, Chengdu, 610064, Sichuan Province, China. 6682631@qq.com.
Xin
X
Yang
School of Life Science, Sichuan University, Animal Disease Prevention and Food Safety Key Laboratory of Sichuan Province, "985 Project" Science Innovative Platform for Resource and Environment Protection of Southwestern, Chengdu, 610064, Sichuan Province, China. whongning@163.com.
Hong-Ning
HN
Wang
School of Life Science, Sichuan University, Animal Disease Prevention and Food Safety Key Laboratory of Sichuan Province, "985 Project" Science Innovative Platform for Resource and Environment Protection of Southwestern, Chengdu, 610064, Sichuan Province, China. 19682292@qq.com.
An-Yun
AY
Zhang
School of Life Science, Sichuan University, Animal Disease Prevention and Food Safety Key Laboratory of Sichuan Province, "985 Project" Science Innovative Platform for Resource and Environment Protection of Southwestern, Chengdu, 610064, Sichuan Province, China. 845701857@qq.com.
Zhi-Kun
ZK
Zhang
School of Life Science, Sichuan University, Animal Disease Prevention and Food Safety Key Laboratory of Sichuan Province, "985 Project" Science Innovative Platform for Resource and Environment Protection of Southwestern, Chengdu, 610064, Sichuan Province, China. 18917573@qq.com.
Wen-Qiao
WQ
Fan
School of Life Science, Sichuan University, Animal Disease Prevention and Food Safety Key Laboratory of Sichuan Province, "985 Project" Science Innovative Platform for Resource and Environment Protection of Southwestern, Chengdu, 610064, Sichuan Province, China. 275484250@qq.com.
Hai-Peng
HP
Cao
D000818:Animals; D000914:Antibodies, Viral / Q000097:blood*; D000956:Antigens, Viral / Q000235:genetics / Q000276:immunology*; D018352:Coronavirus Infections / Q000175:diagnosis / Q000662:veterinary*; D003429:Cross Reactions; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D004926:Escherichia coli / Q000235:genetics; D015870:Gene Expression; D001351:Infectious bronchitis virus / Q000235:genetics / Q000276:immunology*; D008970:Molecular Weight; D011201:Poultry Diseases / Q000175:diagnosis* / Q000821:virology; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics / Q000276:immunology; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D012698:Serologic Tests / Q000379:methods*
D000914:Antibodies, Viral; D000956:Antigens, Viral; D011993:Recombinant Fusion Proteins
Antibodies; ELISA; Escherichia coli; Infectious bronchitis virus; Protein