Ducks as a potential reservoir for Pasteurella multocida infection detected using a new rOmpH-based ELISA

• Published in: Journal of Veterinary Medical Science Vol. 79; no. 7; pp. 1264 - 1271
• Main Authors: LIU, Rongchang, CHEN, Cuiteng, CHENG, Longfei, LU, Ronghui, FU, Guanghua, SHI, Shaohua, CHEN, Hongmei, WAN, Chunhe, LIN, Jiansheng, FU, Qiuling, HUANG, Yu
• Format: Journal Article
• Language: English
• Published: Japan JAPANESE SOCIETY OF VETERINARY SCIENCE 2017; Japan Science and Technology Agency; The Japanese Society of Veterinary Science
• Subjects: Agglutination; Animals; Antibodies; Antisera; Aquatic birds; Bacterial Outer Membrane Proteins - genetics; Bacterial Outer Membrane Proteins - immunology; Bacteriology; China - epidemiology; Cholera; Disease Reservoirs - microbiology; Disease Reservoirs - veterinary; duck; Ducks - microbiology; Enzyme-linked immunosorbent assay; Enzyme-Linked Immunosorbent Assay - methods; Enzyme-Linked Immunosorbent Assay - veterinary; Farms; Fowl cholera; H protein; indirect ELISA; Infections; Membrane proteins; Pasteurella Infections - etiology; Pasteurella Infections - transmission; Pasteurella Infections - veterinary; Pasteurella multocida; Pasteurella multocida - genetics; Pasteurella multocida - immunology; Phylogeny; Poultry Diseases - microbiology; Poultry Diseases - transmission; Poultry farming; Recombinant Proteins; reservoir; rOmpH protein; Sensitivity and Specificity; Waterborne diseases; Western blotting; indirect ELISA; reservoir; rOmpH protein; duck; Pasteurella multocida
• ISSN: 0916-7250; 1347-7439
• DOI: 10.1292/jvms.17-0124

Pasteurella multocida is an important pathogen of numerous domestic poultry and wild animals and is associated with a variety of diseases including fowl cholera. The aim of this study was to develop an indirect enzyme-linked immunosorbent assay (ELISA) based on recombinant outer-membrane protein H (rOmpH) for detection of anti-P. multocida antibodies in serum to determine their prevalence in Chinese ducks. The P. multocida ompH gene was cloned into pET32a, and rOmpH was expressed in Escherichia coli BL21 (DE3). Western blotting revealed that purified rOmpH was recognized by duck antisera against P. multocida, and an indirect ELISA was established. During analysis of serum samples (n=115) from ducks, the rOmpH ELISA showed 95.0% specificity, 100% sensitivity and a 92.0% κ coefficient (95% confidence interval 0.844–0.997) as compared with a microtiter agglutination test. Among 165 randomly selected serum samples, which were collected in 2015 and originated from six duck farms across Fujian Province, China, anti-P. multocida antibodies were detected in 22.42% of apparently healthy ducks, including 25 of 90 sheldrakes (27.8%), eight of 50 Peking ducks (16.0%) and four of 25 Muscovy ducks (16%). Overall, the data suggest that rOmpH is a suitable candidate antigen for the development of an indirect ELISA for detection of P. multocida in ducks; moreover, our results showed that ducks could serve as a potential reservoir for P. multocida infection.