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

Authors :: Martínez E
Carmelo E
Alonso R
Ortega A
Piñero J
del Castillo A
Valladares B
Source :: Letters in applied microbiology [Lett Appl Microbiol] 2003; Vol. 36 (1), pp. 30-4.
Publication Year :: 2003
Abstract: Aims: To develop a rapid colourimetric assay for the detection of Toxoplasma gondii DNA using polystyrene beads as solid support.<br />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.<br />Conclusions: The PCR-ELISA-beads assay is a valuable tool for the detection of T. gondii DNA.<br />Significance and the 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.

Subjects :: Animals
Blotting, Southern
DNA Probes
Microspheres
Reproducibility of Results
Toxoplasma genetics
DNA, Protozoan analysis
Enzyme-Linked Immunosorbent Assay methods
Polymerase Chain Reaction methods
Polystyrenes chemistry
Toxoplasma isolation & purification
Toxoplasmosis diagnosis

Full Text Access

Tools