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°C and 16 samples (identified as G2/G3 using sequencing) showed a mean T m of 77.0°C. The detected mean difference of the T m of 0.6°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.

Development of a real-time PCR for the differentiation of the G1 and G2/G3 genotypes of Echinococcus granulosus

MAURELLI, MARIA PAOLA;RINALDI, LAURA;CRINGOLI, GIUSEPPE
2009

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°C and 16 samples (identified as G2/G3 using sequencing) showed a mean T m of 77.0°C. The detected mean difference of the T m of 0.6°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.
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Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/11588/358505
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