Detection of PPMV-1 antibodies in pigeons using a blocking ELISA based on an NP-specific monoclonal antibody.

Zhi Zhao, Jianing Huang, Xiaoge Duan, Shuling Meng, Hailan Chen, Changting Li, Jiakang He, Tianchao Wei

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Article in Poultry science, 2026. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Not yet cited in PubMed.

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8 authors.

Zhi ZhaoCollege of Animal Science and Technology, Guangxi Key Laboratory of Animal Breeding, Disease Control and Prevention, Guangxi University, Nanning, 530004, Guangxi, China.

Jianing HuangCollege of Animal Science and Technology, Guangxi Key Laboratory of Animal Breeding, Disease Control and Prevention, Guangxi University, Nanning, 530004, Guangxi, China.

Xiaoge DuanCollege of Animal Science and Technology, Guangxi Key Laboratory of Animal Breeding, Disease Control and Prevention, Guangxi University, Nanning, 530004, Guangxi, China.

Shuling MengCollege of Animal Science and Technology, Guangxi Key Laboratory of Animal Breeding, Disease Control and Prevention, Guangxi University, Nanning, 530004, Guangxi, China.

Hailan ChenCollege of Animal Science and Technology, Guangxi Key Laboratory of Animal Breeding, Disease Control and Prevention, Guangxi University, Nanning, 530004, Guangxi, China.

Changting LiGuangxi Key Laboratory of Veterinary Biotechnology, Guangxi Veterinary Research Institute, Guangxi, Nanning, 530001, China.

Jiakang HeCollege of Animal Science and Technology, Guangxi Key Laboratory of Animal Breeding, Disease Control and Prevention, Guangxi University, Nanning, 530004, Guangxi, China.

Tianchao WeiCollege of Animal Science and Technology, Guangxi Key Laboratory of Animal Breeding, Disease Control and Prevention, Guangxi University, Nanning, 530004, Guangxi, China. Electronic address: tcwei88@126.com.

Funding

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6 · The paper itself

Abstract

Pigeon paramyxovirus type 1 (PPMV-1) is a prevalent infectious agent causing Newcastle disease in pigeons, yet serological methods for detecting PPMV-1-specific antibodies remain limited. In this study, we developed a blocking enzyme-linked immunosorbent assay (ELISA) based on a monoclonal antibody (mAb) targeting the nucleocapsid (NP) protein of PPMV-1. The B-cell epitopes of the NP protein were computationally predicted, and the core antigenic domain was expressed in Escherichia coli and purified to generate specific mAbs. Following hybridoma technology, a high-affinity mAb (6B8) was produced and used to establish the blocking ELISA. Assay conditions were systematically optimized using checkerboard titration, including antigen concentration, serum dilution, incubation time, blocking buffer, and blocking duration. Analysis of known pigeon serum samples established a cut-off value of 26.87% inhibition (PI%), with an analytical sensitivity of 1:400, representing the highest dilution at which all tested positive sera remained detectable. The assay showed no cross-reactivity with other common avian pathogens, except for limited cross-reaction with Newcastle disease virus La Sota strain. Intra- and inter-batch coefficients of variation were all below 10%. Clinical validation using 86 pigeon serum samples demonstrated a concordance rate of 97.67% (84/86) compared with the hemagglutination inhibition (HI) test. In conclusion, this monoclonal antibody-based blocking ELISA provides a sensitive, specific, and reliable tool for large-scale serological surveillance of PPMV-1 in pigeons.

Indexed as

Blocking ELISAMonoclonal antibodyNP proteinPigeonPPMV-1

Identifiers

PMID42617255

DOI10.1016/j.psj.2026.107490

PMCPMC13520992

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