Development of a rapid polymerase chain reaction-ELISA assay using polystyrene beads for the detection of Toxoplasma gondii DNA.
OBJECTIVE
To develop a rapid colourimetric assay for the detection of Toxoplasma gondii DNA using polystyrene beads as solid support.


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.


CONCLUSIONS
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.
Martínez
E
E
Department of Parasitology, Faculty of Pharmacy, niversity of La Laguna, La Laguna, Tenerife, Spain.
Carmelo
E
E
Alonso
R
R
Ortega
A
A
Piñero
J
J
del Castillo
A
A
Valladares
B
B
D000818:Animals; D015139:Blotting, Southern; D015342:DNA Probes; D016054:DNA, Protozoan / Q000032:analysis*; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D008863:Microspheres; D016133:Polymerase Chain Reaction / Q000379:methods*; D011137:Polystyrenes / Q000737:chemistry*; D015203:Reproducibility of Results; D014122:Toxoplasma / Q000235:genetics / Q000302:isolation & purification*; D014123:Toxoplasmosis / Q000175:diagnosis*
D015342:DNA Probes; D016054:DNA, Protozoan; D011137:Polystyrenes