Competition ELISA, using monoclonal antibodies to the transmissible gastroenteritis virus (TGEV) S protein, for serologic differentiation of pigs infected with TGEV or porcine respiratory coronavirus.
Monoclonal antibodies (MAB) to subsite A (25C9) and subsite D (44C11) of the S protein of transmissible gastroenteritis virus (TGEV) were used in a blocking ELISA on fixed TGEV-infected swine testis cells to differentiate sera from pigs experimentally inoculated with either TGEV or porcine respiratory coronavirus (PRCV). Serum samples were obtained from pigs at various intervals from postinoculation day (PID) 0 through at least PID 22 to 40. Eleven-day-old pigs, seronegative for TGEV-neutralizing antibodies at the time of inoculation, were inoculated orally and nasally with either the virulent Miller (M5C) strain or the attenuated Purdue (P115) strain of TGEV, or with the ISU-1 strain of PRCV. Gastroenteritis was observed in 100% of the M5C-TGEV-inoculated pigs; but clinical signs of disease were not observed in either the P115-TGEV- or PRCV-inoculated pigs. Virus-neutralization (VN) antibody titer in sera was determined by use of a plaque-reduction assay. Blocking ELISA antibody titer for subsites A and D was determined from the serum dilution that produced 50% reduction in the absorbance values when it competed with biotinylated MAB 25C9 and 44C11, respectively. In sera from the inoculated pigs, the VN antibody titer began to increase by PID 7 and reached maximum by PID 15 to 16. For pigs inoculated with TGEV M5C, subsite A and subsite D blocking antibody titers in the serum paralleled the VN antibody titer, began to increase after PID 7, and reached maximum by PID 15 to 16.(ABSTRACT TRUNCATED AT 250 WORDS)
Simkins
R A
RA
Ohio Agricultural Research and Development Center, Ohio State University, Wooster 44691.
Weilnau
P A
PA
Van Cott
J
J
Brim
T A
TA
Saif
L J
LJ
D000818:Animals; D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral / Q000097:blood; D003333:Coronaviridae Infections / Q000175:diagnosis / Q000662:veterinary*; D003937:Diagnosis, Differential; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods / Q000662:veterinary*; D005761:Gastroenteritis, Transmissible, of Swine / Q000175:diagnosis* / Q000276:immunology; D012141:Respiratory Tract Infections / Q000175:diagnosis / Q000382:microbiology / Q000662:veterinary*; D013552:Swine; D013553:Swine Diseases / Q000175:diagnosis* / Q000382:microbiology; D005760:Transmissible gastroenteritis virus / Q000276:immunology; D014764:Viral Proteins / Q000276:immunology*
D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral; D014764:Viral Proteins