Modification of gDNA extraction from soil for PCR designed for the routine examination of soil samples contaminated with Toxocara spp. eggs.
A modification of gDNA extraction was developed for the polymerase chain reaction (PCR) technique, intended for the detection and differentiation of Toxocara spp. eggs in soil or sediments. Sand samples from sandpits confirmed as being contaminated with Toxocara spp. eggs by the flotation technique were analysed by PCR. The use of proteinase K made it possible to obtain genomic DNA from the sample without needing to isolate eggs using flotation or to inactivate PCR inhibitors present in the sand. Specific primers in the PCR reaction allowed discrimination between T. canis and T. cati eggs. The modification simplified the procedure, thanks to eliminating the step of gDNA isolation from eggs, which is both laborious and difficult.
Laboratory of Parasitoses of Domestic Animals, Institute of Parasitology of the Polish Academy of Sciences, Twarda 51/55, 00-818 Warsaw, Poland. ab@twarda.pan.pl
A
A
Borecka
J
J
Gawor
D000818:Animals; D018509:DNA, Helminth / Q000032:analysis*; D010270:Parasite Egg Count; D016133:Polymerase Chain Reaction / Q000379:methods*; D012822:Silicon Dioxide; D012987:Soil / Q000469:parasitology*; D014119:Toxocara / Q000235:genetics / Q000302:isolation & purification*
D018509:DNA, Helminth; D012987:Soil; D012822:Silicon Dioxide