Establishment of a peptide-based enzyme-linked immunosorbent assay for detecting antibodies against PRRSV M protein. 2021

Jun Zhao, and Rubo Zhang, and Ling Zhu, and Huidan Deng, and Fengqing Li, and Lei Xu, and Jianbo Huan, and Xiangang Sun, and Zhiwen Xu
College of Veterinary Medicine, Sichuan Agricultural University, Chengdu, 611130, China.

BACKGROUND Porcine reproductive and respiratory syndrome (PRRS) is one of the most economically devastating diseases affecting the swine industry globally. Evaluation of antibody responses and neutralizing antibody titers is the most effective method for vaccine evaluation. In this study, the B cell line epitopes of PRRSV M protein were predicted, and two peptide ELISA assays were established (M-A110-129 ELISA, M-A148-174 ELISA) to detect antibodies against PRRSV M protein. Field serum samples collected from pig farms were used to validate the peptide ELISA and compare it with an indirect immunofluorescence assay. RESULTS The sensitivity and specificity of M-A110-129 ELISA and M-A148-174 ELISA were (111/125) 88.80%, (69/70) 98.57% and (122/125) 97.60%, (70/70) 100%, relative to indirect immunofluorescence assay. This peptide ELISA could detect antibodies against different genotypes of PRRSV including type 1 PRRSV, classical PRRSV, HP-PRRSV, and NADC30 like PRRSV, but not antibodies against other common swine viruses. The results of ROC analysis showed that the area under the curve (AUC) of the M-A110-129 ELISA and M-A148-174 ELISA were 0.967 and 0.996, respectively. Compared the concordance of results using two peptide ELISA assays, the IDEXX PRRSV X3 Ab ELISA and a virus neutralization test, were assessed using a series of 147 sera from pigs vaccinated with the NADC30-like PRRSV inactivated vaccine. The M-A148-174 ELISA had the best consistency, with a Cohen's kappa coefficient of 0.8772. The concordance rates of the Hipra PRRSV ELISA kit, M-A110-129 ELISA and M-A148-174 ELISA in the field seropositive detection results were 91.08, 86.32 and 95.35%, relative to indirect immunofluorescence assay. CONCLUSIONS In summary, compared with M-A110-129 ELISA, the PRRSV M-A148-174 ELISA is of value for detecting antibodies against PRRSV and the evaluation of the NADC30-like PRRSV inactivated vaccine, but the advantage is insufficient in serological early diagnosis.

UI MeSH Term Description Entries
D004797 Enzyme-Linked Immunosorbent Assay An immunoassay utilizing an antibody labeled with an enzyme marker such as horseradish peroxidase. While either the enzyme or the antibody is bound to an immunosorbent substrate, they both retain their biologic activity; the change in enzyme activity as a result of the enzyme-antibody-antigen reaction is proportional to the concentration of the antigen and can be measured spectrophotometrically or with the naked eye. Many variations of the method have been developed. ELISA,Assay, Enzyme-Linked Immunosorbent,Assays, Enzyme-Linked Immunosorbent,Enzyme Linked Immunosorbent Assay,Enzyme-Linked Immunosorbent Assays,Immunosorbent Assay, Enzyme-Linked,Immunosorbent Assays, Enzyme-Linked
D000818 Animals Unicellular or multicellular, heterotrophic organisms, that have sensation and the power of voluntary movement. Under the older five kingdom paradigm, Animalia was one of the kingdoms. Under the modern three domain model, Animalia represents one of the many groups in the domain EUKARYOTA. Animal,Metazoa,Animalia
D012680 Sensitivity and Specificity Binary classification measures to assess test results. Sensitivity or recall rate is the proportion of true positives. Specificity is the probability of correctly determining the absence of a condition. (From Last, Dictionary of Epidemiology, 2d ed) Specificity,Sensitivity,Specificity and Sensitivity
D013552 Swine Any of various animals that constitute the family Suidae and comprise stout-bodied, short-legged omnivorous mammals with thick skin, usually covered with coarse bristles, a rather long mobile snout, and small tail. Included are the genera Babyrousa, Phacochoerus (wart hogs), and Sus, the latter containing the domestic pig (see SUS SCROFA). Phacochoerus,Pigs,Suidae,Warthogs,Wart Hogs,Hog, Wart,Hogs, Wart,Wart Hog
D014763 Viral Matrix Proteins Proteins associated with the inner surface of the lipid bilayer of the viral envelope. These proteins have been implicated in control of viral transcription and may possibly serve as the "glue" that binds the nucleocapsid to the appropriate membrane site during viral budding from the host cell. Membrane Proteins, Viral,Viral M Proteins,Viral M Protein,Viral Membrane Proteins
D015164 Vaccines, Inactivated Vaccines in which the infectious microbial nucleic acid components have been destroyed by chemical or physical treatment (e.g., formalin, beta-propiolactone, gamma radiation) without affecting the antigenicity or immunogenicity of the viral coat or bacterial outer membrane proteins. Inactivated Vaccine,Killed Vaccine,Killed Vaccines,Vaccines, Killed,Inactivated Vaccines,Vaccine, Inactivated,Vaccine, Killed
D057134 Antibodies, Neutralizing Antibodies that reduce or abolish some biological activity of a soluble antigen or infectious agent, usually a virus. Neutralizing Antibodies,Antibody, Neutralizing,Neutralizing Antibody
D019084 Fluorescent Antibody Technique, Indirect A form of fluorescent antibody technique commonly used to detect serum antibodies and immune complexes in tissues and microorganisms in specimens from patients with infectious diseases. The technique involves formation of an antigen-antibody complex which is labeled with fluorescein-conjugated anti-immunoglobulin antibody. (From Bennington, Saunders Dictionary & Encyclopedia of Laboratory Medicine and Technology, 1984) Immunofluorescence Antibody Test, Indirect,Immunofluorescence Technique, Indirect,Fluorescent Antibody Technic, Indirect,Immunofluorescence Technic, Indirect,Indirect Fluorescent Antibody Technic,Indirect Fluorescent Antibody Technique,Indirect Immunofluorescence,Indirect Immunofluorescence Assay,Assay, Indirect Immunofluorescence,Assays, Indirect Immunofluorescence,Immunofluorescence Assay, Indirect,Immunofluorescence Assays, Indirect,Immunofluorescence Technics, Indirect,Immunofluorescence Techniques, Indirect,Immunofluorescence, Indirect,Immunofluorescences, Indirect,Indirect Immunofluorescence Assays,Indirect Immunofluorescence Technic,Indirect Immunofluorescence Technics,Indirect Immunofluorescence Technique,Indirect Immunofluorescence Techniques,Indirect Immunofluorescences
D019316 Porcine respiratory and reproductive syndrome virus A species of ARTERIVIRUS causing reproductive and respiratory disease in pigs. The European strain is called Lelystad virus. Airborne transmission is common. Lelystad Virus,PRRSV,Swine Infertility and Respiratory Syndrome Virus,Porcine Reproductive and Respiratory Syndrome Virus,Swine Infertility Respiratory Syndrome Virus
D019318 Porcine Reproductive and Respiratory Syndrome A syndrome characterized by outbreaks of late term abortions, high numbers of stillbirths and mummified or weak newborn piglets, and respiratory disease in young unweaned and weaned pigs. It is caused by PORCINE RESPIRATORY AND REPRODUCTIVE SYNDROME VIRUS. (Radostits et al., Veterinary Medicine, 8th ed, p1048) Blue-Eared Pig Disease,Mystery Swine Disease,PRRS,Porcine Epidemic Abortion and Respiratory Syndrome,Swine Infertility and Respiratory Syndrome,Blue Eared Pig Disease,Pig Disease, Blue-Eared,Swine Disease, Mystery

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