Detection of Infectious Bursal Disease Virus by Double Antibody Sandwich ELISA

Infectious bursal disease virus (IBDV) is responsible for a highly contagious disease of poultry causing severe immunosuppression in chickens. A double antibody sandwich ELISA (DAS-ELISA) was developed to detect IBDV from clinical samples. Two kinds of anti-IBDV antibodies, monoclonal antibody R63 and chicken anti-IBDV sera, were used for DAS-ELISA. Detection limit of IBDV by DAS-ELISA was approximately 10(2.7) EID(50)/ml. The DAS-ELISA detected IBDV from most (13/14) of vaccine products including mild, intermediate and intermediate-plus types. The DAS-ELISA also detected IBDV from all (19/19) of field Korean isolates including very virulent and intermediate-plus phenotypes. Our results indicate that the DAS-ELISA would provide useful diagnostic tool to detect IBDV from clinical samples as well as rapid quantitative detection of IBDV.

Detection of Antibodies to Infectious Bursal Disease Virus (IBDV) by Agar Gel Immunodiffusion using Recombinant VP2 Protein

Infectious bursal disease virus (IBDV) causes a highly contagious and immunosuppressive disease of chicken. Agar gel immunodiffusion using IBDV antigen extracted from bursa of Fabricius of infected chicken has been used officially for diagnosis of IBDV in Korea. In this study, in order to replace the IBDV whole virus antigen with non-infectious antigen, recombinant VP2 protein (rVP2) of IBDV was produced using recombinant baculovirus expression system. Purified baculovirus-expressed rVP2 was used as an antigen in an agar gel immunodiffusion (AGID). rVP2 antigen precipitated specifically IBDV antibodies. AGID using rVP2 antigen detected anti-IBDV antibodies from 6 dpi to 28 dpi (termination of the experiment) when specific pathogen free chickens were experimentally infected with IBDV 52/70 strain. This was consistent with result by AGID using IBDV antigen, virus neutralization test (VNT) and a commercial ELISA kit (except for one serum). The sensitivity of rVP2 was the same with that of IBDV antigen when field sera (n=324) were tested by AGID. However, AGID using rVP2 antigen detected maternal antibodies from broiler chickens (n=20) on a broiler farm up to 15 days old, although the detection rate of the AGID was relatively low compared to a commercial ELISA kit. Our results indicate that IBDV whole virus antigen from IBDV infected chickens would be replaced with recombinant VP2 protein as an antigen for AGID.

Enzyme-linked immunosorbent assay for detection of Trichinella spiralis antibodies and the surveillance of selected pig breeding farms in the Republic of Korea

Trichinellosis is a parasitic zoonosis of public health importance. It is caused by Trichinella spiralis which has a wide host range including humans. In the present communication, the ELISA technique was employed on a total of 803 blood samples from 7 selected pig breeding farms in 1996 for diagnosis and surveillance of trichinellosis. Out of the entire 803 samples, nine were found to be suspected while one was positive by ELISA. But western blot analyses employed for further confirmation have shown that all of 10 samples did not react to larval excretory-secretory product antigens. These results indicate that pig breeding farms included in the present study are free from trichinellosis. However, it does not mean Korea is free from trichinellosis since human trichinellosis has recently been reported. The necessity of continued surveillance for trichinellosis in both pigs and wild animals was discussed.

Toxoplasma antibody titers by ELISA and indirect latex agglutination test in pregnant women

The seroepidemiologic studies on anti-Toxoplasma antibody titers were carried out using ELISA and indirect latex agglutination test. Among 899 sera prepared from pregnant women, 39 cases (4.3%) revealed positive reaction and 218 sera from middle school students showed 4 positive reaction (1.8%) by ELISA. By LAT(newly established by National Veterinary Research Institute, Korea), the sera of 7 pregnant women (0.8%) showed positive reaction. When 80 sera showing +S1:8 by LAT were used for comparing the results obtained from LAT and Toxotest-MT (Eiken Chemical Co., Japan), 7 cases and 8 sera were positive, respectively. All of 11 sera of proven toxoplasmosis patients showed positive reaction in both tests. Overall proportion of agreement between LAT kit and Toxotest-MT was 0.94( -index= 0.632, p<0.01), and LAT was considered to be useful for the screening of toxoplasmosis.