[Expression of GP5-M fusion protein of porcine reproductive and respiratory syndrone virus (PRRSV) and establishment of ELISA diagnose based on the recombinant fusion protein].
The cDNA fragment encoding the truncated GP5 and the full-length M protein of Porcine Reproductive and Respiratory Syndrone Virus (PRRSV) were orderly fused to the downstream of glutathione S-transferase (GST) of pGEX-KG expression vector, resulting in the fusion expression plasmid pKG-56. After transformed into E. coli BL21 (DE3) and induced by IPTG, the results of SDS-PAGE showed that the GST-GP5-M fusion protein was expressed in high level. Western-blot was performed to confirm that the expressed fusion protein could specifically react with antiserum against PRRSV. The fusion protein was further purified and used as an antigen to establish a novel PRRSV ELISA diagnose assay (P56-ELISA). Comparison between P56-ELISA and the abroad kit IDEXX-ELISA showed the two methods had 94.1 percent agreement by detecting 205 serum samples, indicating that the indirect P56-ELISA was specific and sensitive. The correlation between virus neutralization antibody of the infected pigs (not convalescent pigs) and antibody response to the fusion protein GP5-M was further studied. The regression function analysis suggested that there was no significant correlation between ELISA antibody response (OD630 nm) to the fusion protein GP5-M in clinical serum and their specific neutralizing titers.
Jiang
Yun-Bo
YB
Laboratory of Animal Virology, College of Animal Science and Veterinary Medicine, Huazhong Agriculture University, Wuhan 430070, China.
Fang
Liu-Rong
LR
Xiao
Shao-Bo
SB
Xie
Tian-Tian
TT
Chen
Huan-Chun
HC
D000818:Animals; D004797:Enzyme-Linked Immunosorbent Assay; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D005982:Glutathione Transferase / Q000378:metabolism; D016366:Open Reading Frames; D019318:Porcine Reproductive and Respiratory Syndrome / Q000175:diagnosis* / Q000276:immunology; D019316:Porcine respiratory and reproductive syndrome virus / Q000235:genetics* / Q000276:immunology; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis* / Q000235:genetics; D013552:Swine; D014759:Viral Envelope Proteins / Q000096:biosynthesis* / Q000235:genetics
D011993:Recombinant Fusion Proteins; D014759:Viral Envelope Proteins; C114789:glycoprotein 5, PRRSV; D005982:Glutathione Transferase