Development of an epitope-based competitive ELISA for the detection of antibodies against Tibetan peste des petits ruminants virus.
OBJECTIVE
To develop an effective diagnostic kit, based on a competitive ELISA-based system (cELISA), for detecting serum antibody against peste des petits ruminants virus (PPRV).


METHODS
Epitope peptides of the nucleocapsid (N) protein of Tibetan PPRV were synthesized chemically and injected into rabbits to prepare hyperimmune antisera. Test sera were incubated simultaneously with hyperimmune antisera and added to the wells of ELISA plates coated previously with recombinant N protein. Horseradish peroxidase-conjugated goat anti-rabbit antibody was employed to detect the quantity of hyperimmune antisera combined with recombinant N protein.


RESULTS
A cELISA has been developed for monitoring PPRV infections with a cutoff value of 35. Relative sensitivity and specificity values of the epitope-based cELISA were 96.18 and 91.29%, respectively, when compared with a commercial cELISA kit in a test involving 1,039 serum samples.


CONCLUSIONS
We report an efficient method for preparing antibody suitable for incorporation into a cELISA that can be used routinely for the detection of PPRV antibodies in serum samples. The method eliminated the requirement for virus culture and monoclonal antibody preparation, reduced the biorisk posed by virus-dependent manipulations, and the performance of the resultant cELISA compared favorably with a commercially available cELISA kit.
Institute of Animal Sciences and Veterinary Medicine and Shanghai Key Laboratory of Agricultural Genetics and Breeding, Shanghai Academy of Agricultural Sciences, SAAS, Shanghai, China.
Guo-Rui
GR
Zhang
Rui-Song
RS
Yu
Jiang-Yong
JY
Zeng
Yu-Min
YM
Zhu
Shi-Juan
SJ
Dong
Luobu
L
Dunzhu
Se
S
Zhu
Ciren
C
Duoji
Zhi-Hai
ZH
Lei
Zhen
Z
Li
D000818:Animals; D000914:Antibodies, Viral / Q000097:blood*; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods / Q000662:veterinary*; D015511:Goat Diseases / Q000175:diagnosis* / Q000276:immunology / Q000821:virology; D006041:Goats; D019590:Nucleocapsid Proteins / Q000276:immunology*; D029021:Peste-des-Petits-Ruminants / Q000175:diagnosis / Q000276:immunology / Q000662:veterinary*; D018107:Peste-des-petits-ruminants virus / Q000276:immunology*; D011817:Rabbits; D012756:Sheep; D012757:Sheep Diseases / Q000175:diagnosis* / Q000276:immunology / Q000821:virology
D000914:Antibodies, Viral; D019590:Nucleocapsid Proteins