Development of a rapid polymerase chain reaction-ELISA assay using polystyrene beads for the detection of Toxoplasma gondii DNA

Abstract

Aims: To develop a rapid colourimetric assay for the detection of Toxoplasma gondii DNA using polystyrene beads as solid support. Methods and Results: A nested-polymerase chain reaction (PCR)-ELISA assay for the detection of T. gondii DNA was standardized by optimizing the hybridization time and probe concentration. Its detection threshold was then determined and compared with Southern blotting hybridization. These were found to be equivalent, but the PCR-ELISA-beads test is easier to perform and the turnaround time is much shorter than with Southern blot. Conclusions: The PCR-ELISA-beads assay is a valuable tool for the detection of T. gondii DNA. Significance and Impact of the Study: Our results demonstrate that this PCR-ELISA assay, using polystyrene beads, can be used as a routine diagnostic test for the detection of T. gondii in clinical laboratories.