Random amplified polymorphic DNA for the specific detection of bubaline Echinococcus granulosus by hybridization assay

Abstract The polymerase chain reaction (PCR) method to randomly amplify polymorphic DNA (RAPD) was used to differentiate the bubaline and bovine strains of Echinococcus granulosus and buffalo host DNA. Four random oligonucleotide primers of 10–11 mer were analyzed for their ability to direct the amplification of polymorphic DNA fragments from parasites and bovine DNA. Significant DNA polymorphism was observed between the E. granulosus isolates. A selectively amplified DNA fragment 0.9 kilobase (kb) from E. granulosus buffalo isolate by primer AP2 (5′-TGCCGAGCTG-3′) was reamplified and used as a DIG-labelled DNA probe. Dot-blot hybridization of total genomic DNA differentiated buffalo E. granulosus isolate from bovine isolate and bubaline host DNA with no detectable cross-hybridization signal.