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