Development and evaluation of the rVP-ELISA for detection of antibodies against porcine parvovirus

Miaomiao Kong; Yonggang Peng; Yuchao Cui; Tiecheng Chang; Xiaoling Wang; Zhaoxia Liu; Yonggang Liu; Yu Zhu; Yakun Luo; Qinghai Tang; Li Feng; Shangjin Cui

doi:10.1016/j.jviromet.2014.06.008

Development and evaluation of the rVP-ELISA for detection of antibodies against porcine parvovirus

J Virol Methods. 2014 Sep:206:115-8. doi: 10.1016/j.jviromet.2014.06.008. Epub 2014 Jun 16.

Authors

Miaomiao Kong¹, Yonggang Peng², Yuchao Cui³, Tiecheng Chang³, Xiaoling Wang², Zhaoxia Liu², Yonggang Liu², Yu Zhu⁴, Yakun Luo¹, Qinghai Tang⁵, Li Feng², Shangjin Cui⁶

Affiliations

¹ State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute of Chinese Academy of Agricultural Sciences, 427 Maduan Street, Nangang District, Harbin 150001, Heilongjiang, China; Institute of Animal Sciences (IAS), Chinese Academy of Agricultural Sciences, Beijing 100094, China.
² State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute of Chinese Academy of Agricultural Sciences, 427 Maduan Street, Nangang District, Harbin 150001, Heilongjiang, China.
³ State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute of Chinese Academy of Agricultural Sciences, 427 Maduan Street, Nangang District, Harbin 150001, Heilongjiang, China; College of Animal Science and Technology, HLJ August First Land Reclamation University, Daqing 163319, China.
⁴ State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute of Chinese Academy of Agricultural Sciences, 427 Maduan Street, Nangang District, Harbin 150001, Heilongjiang, China; Institute of Animal Sciences (IAS), Chinese Academy of Agricultural Sciences, Beijing 100094, China; College of Animal Science and Technology, HLJ August First Land Reclamation University, Daqing 163319, China.
⁵ Nanyang Normal University, 1638 Wolong Road, Nanyang 473061, China.
⁶ State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute of Chinese Academy of Agricultural Sciences, 427 Maduan Street, Nangang District, Harbin 150001, Heilongjiang, China; Institute of Animal Sciences (IAS), Chinese Academy of Agricultural Sciences, Beijing 100094, China; College of Animal Science and Technology, HLJ August First Land Reclamation University, Daqing 163319, China. Electronic address: cuishangjin@126.com.

PMID: 24945904
DOI: 10.1016/j.jviromet.2014.06.008

Abstract

The gene encoding the VP2 protein of porcine parvovirus (PPV) was expressed in an insect-baculovirus system. The recombinant (r) VP2 was similar antigenically/functionally to the native capsid protein as demonstrated by hemagglutination (HA), Western blotting using PPV positive sera. The purified rVP2 proteins were used as coating antigen to establish a rVP-ELISA method for detection of PPV positive and negative sera from pigs. The optimal operating conditions of the rVP-ELISA were: the concentration of rVP2 proteins coated on the wells was 2 μg/mL; the diluted concentration of serum was 1: 150 and that of the enzyme-labeled antibody was 1: 6000. A total of 596 sera were detected by this assay, and the average positive rate was 87%. Compared with France LSI kit, the result showed that the coincidence rate was 96.7%. In conclusion, the rVP2-ELISA is a sensitive and specific method for detecting antibodies against PPV.

Keywords: Insect-baculovirus system; Porcine parvovirus; rVP-ELISA; rVP2.

Publication types

Evaluation Study
Research Support, Non-U.S. Gov't

MeSH terms

Animals
Antibodies, Viral / blood*
Antigens, Viral* / genetics
Baculoviridae / genetics
Capsid Proteins* / genetics
Clinical Laboratory Techniques / methods
Enzyme-Linked Immunosorbent Assay / methods
Genetic Vectors
Parvoviridae Infections / diagnosis
Parvoviridae Infections / veterinary*
Parvoviridae Infections / virology
Parvovirus, Porcine / immunology*
Parvovirus, Porcine / isolation & purification
Sensitivity and Specificity
Sf9 Cells
Spodoptera
Swine
Swine Diseases / diagnosis*
Swine Diseases / virology
Veterinary Medicine / methods*

Substances

Antibodies, Viral
Antigens, Viral
Capsid Proteins
VP2 protein, porcine parvovirus