Hostname: page-component-586b7cd67f-t7fkt Total loading time: 0 Render date: 2024-11-28T16:19:24.976Z Has data issue: false hasContentIssue false
  • English
  • Français

Modification of gDNA extraction from soil for PCR designed for the routine examination of soil samples contaminated with Toxocara spp. eggs

Published online by Cambridge University Press:  01 June 2008

A. Borecka  and
J. Gawor
A. Borecka*
Affiliation:
Laboratory of Parasitoses of Domestic Animals, Institute of Parasitology of the Polish Academy of Sciences, Twarda 51/55, 00-818Warsaw, Poland
J. Gawor
Affiliation:
Laboratory of Parasitoses of Domestic Animals, Institute of Parasitology of the Polish Academy of Sciences, Twarda 51/55, 00-818Warsaw, Poland
*
*E-mail: [email protected]
Get access Rights & Permissions [Opens in a new window]

Abstract

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.

Type
Research Papers
Information
Journal of Helminthology , Volume 82 , Issue 2 , June 2008 , pp. 119 - 122
Copyright
Copyright © Cambridge University Press 2008

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)

References

Borecka, A. (2004) Differentiation of Toxocara spp. eggs isolated from the soil by PCR-linked RFLP. Helminthologia 41, 185–187.Google Scholar
Dada, B.J.O. (1979) A new technique for the recovery of Toxocara eggs from soil. Journal of Helminthology 53, 141–144.CrossRefGoogle ScholarPubMed
Dubna, S., Langrova, I., Jankovska, I., Vadlejch, J., Pekar, S., Napravnik, J. & Fechtner, J. (2007) Contamination of soil with Toxocara eggs in urban (Prague) and rural areas in the Czech Republic. Veterinary Parasitology 144, 81–86.CrossRefGoogle ScholarPubMed
Fogt-Wyrwas, R., Jarosz, W. & Mizgajska-Wiktor, H. (2007) Utilizing a polymerase chain reaction method for the detection of Toxocara canis and T. cati eggs in soil. Journal of Helminthology 81, 75–78.CrossRefGoogle Scholar
Gawor, J. & Borecka, A. (2004) The contamination of the environment with Toxocara eggs in Mazowieckie voivodship as a risk of toxocarosis in children. Wiadomości Parazytologiczne 50, 237–241.Google ScholarPubMed
Giacometti, A., Cirioni, O., Fortuna, M., Osimani, P., Antonicelli, L., del Prete, M.S., Riva, A., D'Errico, M.M., Petrelli, E. & Scalise, G. (2000) Environmental and serological evidence for the presence of toxocariasis in the urban area of Ancona, Italy. European Journal of Epidemiology 16, 1023–1026.CrossRefGoogle ScholarPubMed
Halos, L., Jamal, T., Vial, L., Maillard, R., Suau, A., Le Menach, A., Boulouis, H.J. & Vayssier-Taussat, M. (2004) Determination of an efficient and reliable method for DNA extraction from ticks. Veterinary Research 35, 709–713.CrossRefGoogle ScholarPubMed
Jacobs, D.E., Zhu, X., Gasser, R.B. & Chilton, N.B. (1997) PCR-based methods for identification of potentially zoonotic ascaridoid parasites of the dog, fox and cat. Acta Tropica 68, 191–200.CrossRefGoogle ScholarPubMed
Kulišić, Z., Pavlović, I., Milutinović, M. & Aleksić-Bakrać, N. (1998) Internal parasites of dogs and role of dogs in epidemiology of larva migrans in the Belgrade area. Helminthologia 35, 79–82.Google Scholar
Moreira, D. (1998) Efficient removal of PCR inhibitors using agarose-embedded DNA preparations. Nucleic Acids Research 26, 3309–3310.CrossRefGoogle ScholarPubMed
Muradian, V., Gennari, S.M., Glickman, L.T. & Pinheiro, S.R. (2005) Epidemiological aspects of visceral larva migrans in children living at São Paulo (SP), Brasil. Veterinary Parasitology 134, 93–97.CrossRefGoogle Scholar
O'Lorcain, P. (1994) Epidemiology of Toxocara spp. in stray dogs and cats in Dublin, Ireland. Journal of Helminthology 68, 331–336.CrossRefGoogle ScholarPubMed
Saeki, H., Yokoi, H. & Yamamoto, M. (1997) Long term survey on intestinal nematode and cestode infections in stray puppies Ibaraki Prefecture. Journal of Veterinary Medical Science 59, 725–726.Google ScholarPubMed
Uga, S., Matsuo, J., Kimura, D., Rai, S.K., Koshino, Y. & Igarashi, K. (2000) Differentiation of Toxocara canis and T. cati eggs by light and scanning electron microscopy. Veterinary Parasitology 92, 287–294.CrossRefGoogle Scholar
Wu, Z., Nagano, I., Xu, D. & Takahashi, Y. (1997) Primers for polymerase chain reaction to detect genomic DNA of Toxocara canis and T. cati. Journal of Helminthology 71, 77–78.CrossRefGoogle ScholarPubMed
Yeates, C., Gillings, M.R., Davison, A.D., Altavilla, N. & Veal, D.A. (1998) Methods for microbial DNA extraction from soil for PCR amplification. Biological Procedures Online 1, 40–47.CrossRefGoogle ScholarPubMed