Development of a lateral flow assay for rapid and accurate detection of chicken anemia virus
1
Issued Date
2024-03-01
Resource Type
ISSN
00325791
eISSN
15253171
Scopus ID
2-s2.0-85182786065
Journal Title
Poultry Science
Volume
103
Issue
3
Rights Holder(s)
SCOPUS
Bibliographic Citation
Poultry Science Vol.103 No.3 (2024)
Suggested Citation
Angsujinda K., Peala W., Sittidech A., Wanganurakkul S., Mahony T.J., Wang S.F., Smith D.R., Chintapitaksakul L., Khongchareonporn N., Assavalapsakul W. Development of a lateral flow assay for rapid and accurate detection of chicken anemia virus. Poultry Science Vol.103 No.3 (2024). doi:10.1016/j.psj.2024.103432 Retrieved from: https://repository.li.mahidol.ac.th/handle/123456789/95860
Title
Development of a lateral flow assay for rapid and accurate detection of chicken anemia virus
Corresponding Author(s)
Other Contributor(s)
Abstract
Significant challenges to poultry health are posed by chicken anemia virus (CAV), which induces immunosuppression and causes increased susceptibility to secondary infections. The effective management and containment of CAV within poultry stocks require precise and prompt diagnosis. However, a deficiency persists in the availability of low-cost, rapid, and portable CAV detection devices. In this study, an immunochromatographic lateral-flow test strip-based assay was developed for CAV detection using in-house generated monoclonal antibodies (MABs) against CAV viral protein 1 (VP1). The recombinant truncated VP1 protein (Δ60VP1), with amino acid residues 1 to 60 of the native protein deleted, was produced via a prokaryotic expression system and utilized for immunizing BALB/c mice. Subsequently, high-affinity MABs against Δ60VP1 were generated and screened using conventional hybridoma technology combined with serial dilution assays. Two MABs, MAB1, and MAB3, both binding to distinct epitopes of Δ60VP1, were selected for the development of a lateral-flow assay. Sensitivity analysis demonstrated that the Δ60VP1 antigen could be detected by our homemade lateral-flow assay at concentrations as low as 625 ng/mL, and this sensitivity was maintained for at least 6 mo. The assay exhibited high specificity, as evidenced by its lack of reactivity with surrogate recombinant proteins and the absence of cross-reactivity with other chicken viruses and viral antigens. Comparative analysis with quantitative PCR data demonstrated substantial agreement, with a Kappa coefficient of 0.66, utilizing a sample set comprising 305 clinical chicken serum samples. In conclusion, the first lateral-flow assay for CAV detection was developed in this study, utilizing 2 specific anti-VP1 MABs. It is characterized by simplicity, rapidity, sensitivity, and specificity.