Molecular appraisal of Indian animal isolates of Echinococcus granulosus

Abstract

Background & objective: Cystic echinococcosis (CE) has a wide host range and distinct entities, not only reflected phenotypically but also by genotypic variation. Considering this fact, this study was undertaken to characterize the Indian isolates of Echinococcus granulosus to find out difference between Indian cattle, buffalo and sheep isolates on the basis of random amplification of polymorphic DNA (RAPD) PCR and PCR mediated restriction fragment length polymorphism (PCRRFLP) of internal transcribed spacer gene 1 (ITS1).

Methods: A total of 22 isolates of E. granulosus obtained from Indian cattle, buffalo and sheep (December 2004 - November 2005) were analysed by 26 random primers of 8-10 mers. After isolation of protoscoleces from fertile cyst, DNA was extracted, quantified and amplified by random primers. Internal transcribed spacer gene 1 (ITS1) was amplified using specific primer and digested by two restriction enzymes (Msp1 and Rsa1).

Results: Of the 26 primers, only two primers (5'ACC TGG ACA C3' and 5' TCA TCC GAG G3') could discriminate cattle, buffalo and sheep isolates collected from eastern part of India. Samples were further analysed by PCR mediated RFLP of internal transcribed spacer gene1 (ITS1) using two restriction enzymes (Msp1 and Rsa1). No ITS1 variants could be detected.

Interpretation & conclusion: Our findings showed genotypic variation among Indian animal isolates of E. granulosus on the basis of RAPD fingerprinting.