Genotyping of Echinococcus granulose in Samples of Iraqi Patients

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 Iraq. 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. The aim of study was to ascertain the genotype(s) of the parasite responsible for human hydatidosis inIraq. Patients and Methods: After collecting hydatid cysts from patients after surgical operation , DNA samples were extracted from (60) hydatid cyst , 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 2% agarose gels For genotyping of echinoccocus 5 ng template DNA were added into the PCR mix. And 11 primer pair were used for genotyping . Result: Most of the CE patients(60) patients were found to be infected with hydatid cyst of eitherG4 26(43%),G1/G/G3 18(30%), G5 12(20%),G6/G7 4(7%). And the study the association of genotypes and organ ,our results show In the liver (20%) G1/G/G3 ,(27%)G4 ,(7%) G5 ,(20%) G6/G7. Lung (7%) G1/G/G3,(10%)G4,Liver and lung (3%) G1/G/G3 andG4 ,finally liver ,lung and spleen (3%) G1/G/G3. Conclusion: These findings demonstrate the zoonotic potential of G4 (Horse strain ), G1 (sheep strain) and G3 (buffalo strain),G5(Cattle Strain ) , (G6) (Camel Strain ) ,(G7) (Pig strain) genotypes of E. granulosus as these emerged as predominant genotypes infecting the humans in Iraq. Key word: Echinococcus granulosus , PCR , Germinal layer , genotypes .