Evaluation of four DNA extraction methods for the detection of Echinococcus granulosus genotype 1

ABSTRACT

The aim of this survey was to compare four DNA extraction methods from Iranian sheep stain E.granulosus isolates. Cystic echinococcosis [CE] caused by the metacestode of the dog tapeworm Echinococcus spp., is a global zoonotic infection which is economically important and constitutes a major threat to public health in many countries. Strains characterization is essential for the establishment of a preventive and control strategy in every endemic area. Forty five infected organs from cattle, sheep and goat were collected from different abattoirs of Iran. All cysts were examined by microscopic observation of protoscoleces. For each cyst, protoscoleces were aspirated and DNA of each cyst was extracted with 4 different methods including tissue Kit extraction, modified Cinnagen extraction kit, Phenol-chloroform [Sambrook 1999] and modified Phenol chloroform methods. Efficiency of the DNA was determined by degree of success in PCR amplification. Cinnagen modified extraction and modified Phenol chloroform methods were equally effective and superior to other methods after DNA electrophoresis and PCR reaction. Inhibition was observed in PCR with DNA isolated from protoscoleces, and a 1/100 dilution was able to alleviate this problem with DNA extracted. The result of this study show that the quality of extracted DNA using modified Cinnagen extraction kit and modified phenol-chloroform are very high and gave identical results after RCR reaction using 12S rRNA gene. Further evaluation is required for its utilization in other clinical specimens