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Research Note: A novel peptide-based ELISA for efficient detection of antibody against chicken infectious anemia virus.

ABSTRACT: Chicken infectious anemia virus (CIAV) is the pathogen of chicken infectious anemia. Currently, due to the lack of effective diagnostics technology and prevention approach, CIAV has spread globally and caused huge economic losses to poultry industry. In this study, a novel peptide-based ELISA (pELISA) for efficient detection of antibody against CIAV was developed. The peptide (25CRLRRRYKFRHRRRQRYRRRAF45) used in pELISA was highly conserved in VP1 protein of different CIAV isolates. The specificity and reproducibility showed that the pELISA only reacted with sera against CIAV, not with sera against other pathogens tested, and the CV of the intra-/inter-assay of the pELISA was 6.8 to 9.22%. Moreover, the comparison assay using 56 clinical samples showed that the positive rate of the pELISA and the commercial ELISA kit (IDEXX) was 85.7 and 80.4%, respectively. The pELISA generated here provides a rapid and efficient serological detection method for diagnosis of CIAV infection and evaluation of the efficacy of CIAV vaccination.

SUBMITTER: Shao H

PROVIDER: S-EPMC9673107 | biostudies-literature | 2023 Jan

REPOSITORIES: biostudies-literature

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Research Note: A novel peptide-based ELISA for efficient detection of antibody against chicken infectious anemia virus.

Shao Hongxia H Li Jinzhi J Zhang Jun J Zhang Qi Q Ma Li L Lu Jinhua J Li Tuofan T Xie Quan Q Wan Zhimin Z Qin Aijian A Ye Jianqiang J

Poultry science 20221023 1

Chicken infectious anemia virus (CIAV) is the pathogen of chicken infectious anemia. Currently, due to the lack of effective diagnostics technology and prevention approach, CIAV has spread globally and caused huge economic losses to poultry industry. In this study, a novel peptide-based ELISA (pELISA) for efficient detection of antibody against CIAV was developed. The peptide (25CRLRRRYKFRHRRRQRYRRRAF45) used in pELISA was highly conserved in VP1 protein of different CIAV isolates. The specifici ...[more]

PMID: 36399931

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Project description:Avian coronavirus-infectious bronchitis virus (AvCoV-IBV) is the causative agent of infectious bronchitis (IB) that has brought great threat and economic losses to the global poultry industry. Rapid and accurate diagnostic methods are very necessary for effective disease monitoring. At the present study, we screened a novel nanobody against IBV-N protein for development of a rapid, simple, sensitive, and specific competitive ELISA for IBV antibody detection in order to enable the assessment of inoculation effect and early warning of disease infection. Using the phage display technology and bio-panning, we obtained 7 specific nanobodies fused with horseradish peroxidase (HRP) which were expressed in culture supernatant of HEK293T cells. Out of which, the nanobody of IBV-N-Nb66-vHRP has highly binding with IBV-N protein and was easily blocked by the IBV positive serums, which was finally employed as an immunoprobe for development of the competitive ELISA (cELISA). In the newly developed cELISA, we reduce the use of enzyme-conjugated secondary antibody, and the time of whole operation process is approximately 1 h. Moreover, the IBV positive serums diluted at 1:1000 can still be detected by the developed cELISA, and it has no cross reactivity with others chicken disease serums including Newcastle disease virus, Fowl adenovirus, Avian Influenza Virus, Infectious bursal disease virus and Hepatitis E virus. The cut-off value of the established cELISA was 36%, and the coefficient of variation of intra- and inter-assay were 0.55-1.65% and 2.58-6.03%, respectively. Compared with the commercial ELISA (IDEXX kit), the agreement rate of two methods was defined as 98% and the kappa value was 0.96, indicating the developed cELISA has high consistency with the commercial ELISA. Taken together, the novel cELISA for IBV antibody detection is a simple, rapid, sensitive, and specific immunoassay, which has the potential to rapidly test IBV antibody contributing to the surveillance and control of the disease.

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Research Note: A novel peptide-based ELISA for efficient detection of antibody against chicken infectious anemia virus.

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Research Note: A novel peptide-based ELISA for efficient detection of antibody against chicken infectious anemia virus.

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