A recombinant protein-based ELISA for detecting antibodies to foot-and-mouth disease virus serotype Asia 1.
A recombinant protein-based ELISA was evaluated for detecting antibodies to foot-and-mouth disease virus (FMDV) serotype Asia 1. The recombinant protein (rP13C) was derived from the P1 precursor and 3C protease genes that were cloned into a single expression vector and expressed in insect cells. This protein elicited a low titer of FMDV neutralizing antibodies in pigs. Its utility as a diagnostic antigen was explored in a blocking ELISA using monoclonal antibodies. The rP13C ELISA yielded higher endpoint titers than the liquid phase blocking (LPB) ELISA and virus neutralization test performed on sera from goats challenged with FMDV post-vaccination. The rP13C ELISA correctly scored the FMD international reference weak positive serum. The relative sensitivity between the rP13C ELISA and LPB ELISA was equivalent for vaccinated sera. With this comparable sensitivity, the rP13C ELISA exhibited a specificity of 99.7% for domestic naive swine, bovine and caprine sera. This report demonstrates that an ELISA using recombinant proteins has the potential to replace the LPB ELISA using an inactivated FMDV antigen as a simple and robust serological tool for screening antibodies to FMDV serotype Asia 1.
Ko
Young-Joon
YJ
National Veterinary Research and Quarantine Service, Anyang-6-dong, Anyang City, Gyeonggi Province, 430-824, Republic of Korea. koyj@nvrqs.go
Jeoung
Hye-Young
HY
Lee
Hyang-Sim
HS
Chang
Byung-Sik
BS
Hong
Seung-Min
SM
Heo
Eun-Jeong
EJ
Lee
Kwang-Nyeong
KN
Joo
Hoo-Don
HD
Kim
Su-Mi
SM
Park
Jong-Hyeon
JH
Kweon
Chang-Hee
CH
D000086042:3C Viral Proteases; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D000914:Antibodies, Viral / Q000302:isolation & purification*; D036022:Capsid Proteins / Q000276:immunology; D002417:Cattle; D002460:Cell Line; D003546:Cysteine Endopeptidases / Q000096:biosynthesis / Q000276:immunology; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D005536:Foot-and-Mouth Disease / Q000821:virology*; D029544:Foot-and-Mouth Disease Virus / Q000276:immunology / Q000302:isolation & purification*; D006041:Goats; D051379:Mice; D009500:Neutralization Tests; D011817:Rabbits; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000276:immunology; D012680:Sensitivity and Specificity; D018411:Spodoptera; D013552:Swine; D014764:Viral Proteins / Q000096:biosynthesis / Q000276:immunology
D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral; D036022:Capsid Proteins; D011994:Recombinant Proteins; C442501:VP1 protein, Foot-and-mouth disease virus; D014764:Viral Proteins; D003546:Cysteine Endopeptidases; D000086042:3C Viral Proteases