Evaluation of the stability of coated plates with antigen at different temperatures and times by ELISA test to diagnose fasciolosis

ABSTRACT

Considering that ELISA method presently is the test of choice for diagnosis of fasciolosis, the present study was undertaken to evaluate the maximum validity of coated plates at different temperatures and different times during one year of evaluation. Serum samples of patients infected with fasciolosis [n=10], hydatidosis [n=5], toxocariasis [n=5], and negative control sera [n=5] were examined. Two series of plates were considered. The first series were coated with Fasciola homogenate Ag 12 micro g/ml, and after some steps were blocked with gelatin and preserved at different temperatures as -80 °C, -20 °C, -4 °C and +4°C. The 2[nd] series were treated under the same criteria but were not blocked with gelatin. Each series were examined by ELISA test from 1[st] month to 12[th] month. Sera with 1125 dilution, and peroxidase-conjugated goat anti-human IgG diluted 110000 were considered optimum. To ease reporting the results and due to many similarities only results related to 1[st], 6[th] and 12[th] months were analyzed and sensitivity, specificity plus cut-off were determined for each series separately. Preserving the coated plates, while unblocked at -80°C for 6-8 months is pertinent and functional and in that case, we can be sure the best out put would be applicable