Comparison of Five Simple Methods for DNA Extraction from Echinococcus granulosus Protoscoleces for PCR-Amplification of Ribosomal DNA

Abstract

Background: Cystic hydatid disease is an important zoonosis, affecting humans and animals and is a significant public health and economic problem throughout the world and Iran. Since extraction of DNA from the parasite is a primary and crucial step which has a principal effect on PCR results, in the current study five simple methods for DNA extraction from protoscoleces of Echinococcus granulosus were ap­plied and compared with each other.
Methods: After collecting hydatid cysts from an abattoir, DNA samples were extracted from two cyst isolates from sheep, two from goats and two from camels using five different methods involving the use of glass beads, mechanical grinder, freeze-thaw, boiling and crushing. For all DNA samples ex­tracted, one PCR assay based on amplifying rDNA-ITS1 region was performed and amplicons re­solved on 1.5% agarose gels.
Results: The methods were compared regarding to DNA and PCR bands, time and cost effectiveness and laborious amount. The target DNA was successfully amplified from all samples using all methods produced an expected band size. All methods showed some advantages and disadvantages in PCR gels. The boiling method, which was the most time and cost effectiveness method, achieved the thick­est bands in the PCR following grinder, crushing, freeze-thaw and glass beads.
Conclusion: Boiling and crushing methods were the most suitable methods regarding their amplicon quality, easiness, quickness and cost effectiveness.