Development of a real-time PCR for the differentiation of the G1 and G2/G3 genotypes of Echinococcus granulosus.
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.
Maurelli
Maria Paola
MP
Department of Pathology and Animal Health, University of Naples Federico II, Italy.
Rinaldi
Laura
L
Capuano
Federico
F
Perugini
Anna Giannina
AG
Cringoli
Giuseppe
G
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
D052160:Benzothiazoles; D018509:DNA, Helminth; D004272:DNA, Mitochondrial; D003959:Diamines; D009930:Organic Chemicals; D011804:Quinolines; C098022:SYBR Green I