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Abstract
We have developed an ELISA that employs monoclonal anti-Toxoplasma SAG1 (p30) as the capture antibody to detect T. gondii circulating antigens in patients' serum samples. Using serum spiked with Toxoplasma soluble and with SAG1 recombinant proteins, the detection limits were 31.25 ng/mL and 62.50 ng/mL, respectively. We obtained positive results in 28% (21/75) and 11% (23/206) of probable active and chronic toxoplasmosis serum samples, respectively. Western blot analysis on pooled antigen-positive serum samples showed antigenic bands of molecular weights 25 and 75 kDa from sera of probable active infection and five antigenic bands ranging in size from 26 to 33 kDa from chronic infection sera. This assay would be useful as an initial serum selection step in developing a Toxoplasma antigen detection test and for characterization studies.
ACKNOWLEDGMENTS
This study was funded by the eScience Fund (No 02-01-05-SF0154) from the Malaysian Ministry of Science, Technology, and Innovation (MOSTI); and from USM RU grant No. 1001/CIPPM/8130132. The authors would like to thank Mr. Hossein Haj Ghani for his technical assistance in this study.
Notes
a Based on clinical examination and combined evaluation of anti-Toxoplasma IgM and IgG antibodies using ELISA.
b Significant differences (p < 0.01) were observed between positive rates in active and chronic infections as calculated by Z test for two population proportion.