title: Development of a real-time PCR for the differentiation of the G1 and G2/G3 genotypes of Echinococcus granulosus.
abstract: The present study was aimed at developing a SYBR Green-based real-time polymerase chain reaction (PCR) assay for a rapid differentiation of the genotype G1 from the cluster of genotypes G2/G3 of Echinococcus granulosus, using as marker the 12S mtDNA gene. Eleven hydatid cysts from water buffaloes and 19 from cattle were used. Fourteen samples (identified as G1 using sequencing) showed a mean melting temperature (T (m)) of 76.4 degrees C and 16 samples (identified as G2/G3 using sequencing) showed a mean T (m) of 77.0 degrees C. The detected mean difference of the T (m) of 0.6 degrees C between G1 and G2/G3 genotypes might allow a fast and simple discrimination of these genotypes. In conclusion, the real-time PCR developed in the present study provides a powerful tool for molecular studies on E. granulosus with possibilities for extension to other genotypes using different molecular targets.
authors.affiliation: Department of Pathology and Animal Health, University of Naples Federico II, Italy.
authors.forename: Maria Paola
authors.initials: MP
authors.lastname: Maurelli
authors.forename: Laura
authors.initials: L
authors.lastname: Rinaldi
authors.forename: Federico
authors.initials: F
authors.lastname: Capuano
authors.forename: Anna Giannina
authors.initials: AG
authors.lastname: Perugini
authors.forename: Giuseppe
authors.initials: G
authors.lastname: Cringoli
mesh_terms: D000818:Animals; D052160:Benzothiazoles; D002020:Buffaloes / Q000469:parasitology; D002417:Cattle / Q000469:parasitology; D018509:DNA, Helminth / Q000235:genetics; D004272:DNA, Mitochondrial / Q000235:genetics; D003959:Diamines; D048209:Echinococcus granulosus / Q000145:classification* / Q000235:genetics*; D005838:Genotype; D009930:Organic Chemicals / Q000378:metabolism; D016133:Polymerase Chain Reaction / Q000379:methods*; D011804:Quinolines; D013194:Staining and Labeling / Q000379:methods; D044366:Transition Temperature
chemical_list: D052160:Benzothiazoles; D018509:DNA, Helminth; D004272:DNA, Mitochondrial; D003959:Diamines; D009930:Organic Chemicals; D011804:Quinolines; C098022:SYBR Green I