| ObjectiveTo developed an indirect ELISA for peroxiredoxin (Prx) antigen of Toxoplasma gondii in sera of mice. The ELISA detect peroxiredoxin antigen in human serum.to carry out crowd Toxoplasmosis seroepidemiology survey.MethodsThe chessboard titration was used for selection of the ELISA experimental conditions including the concentration of detected serum, the dilution of anti-serum and the enzyme conjugate. To observe whether the results were affected by blocking, and draw standard curve of protein quantitation. The effectiveness was estimated by sera samples of 30 mice infected with Toxoplasma gondii and 30 mice uninfected detected by the indirect ELISA method. The sensitivity, specificity and reliability were detected. Research methods of a cluster sampling extraction study objects. In the shenchi county and shenchi rural areas,Take a group of residents as the objects of studyand Extract their serum. Collection fasting morning venous blood of 1021 study objects,serum collected by centrifugation. Peroxiredoxin Antigen of Toxoplasma gondii in sera of human was detected by the indirect ELISA method.ResultsThe concentration of detected serum for coating was 1:50, the optimal dilutions of rat anti-serum was 1:100 (Titer 1:128),the dilution of HRP-goat anti-rat IgG was 1:4000. however,blocking agent showed no significant effect on the rest result. The developed indirect ELISA method show high sensitivity, reproducibility and specificity. The detection rate of Prx antigen in 30 mice serum samples of mice infected with Toxoplasma gondii was 100%.Positive rate of different age groups of Toxoplasma IgG antibody in Serum is different, the difference is not statistically significant(χ2趋势=4.195,P=0.522).Positive rates of different gender groups of Toxoplasma IgG antibody in serum is different,the difference is not statistically significant (χ2=0.000, P=0.990). Positive rates of people of different educational level of serum Toxoplasma IgG antibody are different, the difference is not statistically significant(χ2趋势=6.322, P=0.097). Positive rate of different occupations of Toxoplasma IgG antibody in serum is different, the difference is not statistically significant(χ2趋势=8.584, P=0.284). Investigated a total of 1021 people, Positive rate of Gondii Toxoplasma IgG antibody without dog and cats is lower than that with domestic dog and cat. Difference between two groups was significant (χ2=13.157, P=0.004).Positive rate of habit of eating raw egg is higher than that without habit of eating raw egg (χ2=8.417, P=0.004,). Toxoplasma IgG antibody positive rate of frequently eaten raw vegetables and fruits contaminated is higher than that of the occasional habit and that of there is no such habit. The difference has statistical significance (χ2=12.201, P=0.002). In 1021 Questionnaire survey, serum antibody-positive rate of the knife board regularly confuse use is high than that of occasional and never confuse use, the difference has statistical significance (χ2=9.767, P=0.008). When Crafting, Toxoplasma IgG antibody positive rate of the frequent contact with the soil is higher than that of occasional contact and never contacts.The difference has statistical significance(χ2=13.753, P=0.001). Washing hands before meals and after each time or regular cleaning person whose positive rate is less than those of positive rate who occasionally or never washing, The difference has statistical significance(χ2=43.125, P<0.001). Toxoplasma IgG antibody positive rates of special populations are:Clinical immune dysfunction is 20.00%. Blood donors are 17.85%. Slaughterring and meat sales are 27.78%. The difference was not significant. (χ2=1.012, P=0.603).ConclusionsThe indirect ELISA method for peroxiredoxin antigen of Toxoplasma gondii in mice sera was successfully developed. The method is sensitive, specific, stable, It could be used as a diagnostic test for toxoplasmosis and also serves as an important tool for epidemiological investigation. Successfully apply ELISA to detect Peroxiredoxin antigen in human serum and apply to seroepidemiological survey.
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