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KMID : 0359319910310040491
Korean Journal of Veterinary Research
1991 Volume.31 No. 4 p.491 ~ p.500
Use of enzyme - linked immunosorbent assay ( ELISA ) for detection of toxoplasmosis in dogs



Abstract
This study was conducted to detect the serum antibodies in the experimentally toxoplasma infected dogs and street dogs by use the of an enzyme-linked immunosorbent assay (ELISA). And this test was performed on the polystylene microplate by coating with the tachyzoites soluble antigen of T gondii (RH strain), incubated with sera diluted then, added with HPO-conjugated rabbit anti-dog IgG and o-phenylenediamine used as a substrate.
Tachyzoites of T gondii harvested from mouse peritcmeal cavity were purified by 30, 40 and 50% Percoll density gradient centrifugation and used as the source of antigen. The results obtained were summarized as follows;
1. The highest ratio of positive to negative (P/N ratio) was obtained at the level of l§¶/§¢ protein concentration of antigen with the 1/4000 dilution of the conjugate measured by checker-board titration. It was regarded as the optimum concentration of the antigen and conjugate.
2. Cut-off value in this IgG ELISA was 0.375 that wa.s determined by mean absorbance (at 492§¬) of IFA negative serum added with the dauble value of the standard deviation (mean¡¾2 S.D.).
3. Serum ELISA IgG antibodies to T gondii in the exyerimentally infected dogs were detected firstly at the Week 3 after inoculation and the highest titer was recognized at the Week 4, 5 and 6 after inoculation.
4. Stability of the antigen absorbed in the microplates that were preserved at 4¡É and -25¡É separately were prolonged up to 3 weeks and 10 weeks at 4¡É and -25¡É, respectively. However the reproducibility was not reliable after the preservation of 4 weeks and longer.
5. Positive rate of the specific antibodies in 312 test sera was 28.5% and there was no significant differences between the male (27.8%) and female (29.5%), respectively.
6. The IgG ELISA was proved to be a specific procedure for the detection of antibodies to canine toxoplasma infection and also evaluated as a screening test for the large scale of test samples in laboratory.
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