Detection of genetic polymorphism among and within Echinococcus granulosus strains by heteroduplex analysis of a microsatellite from the U1 snRNA genes.

RORATTO, P. A.; BARTHOLOMEI-SANTOS, M. L.; GUTIERREZ, A.; KAMENETZKY, L.; MARA CECILIA ROSENZVIT; ZAHA, A.

GENETICS AND MOLECULAR RESEARCH

FUNPEC

Año: 2006 vol. 5 p. 542 - 552

1676-5680

Polymerase chain reaction of a pentanucleotide microsatellite in the U1 snRNAgene complex generated a multiple band pattern due to the priming of paralogoussequences. Denaturation and slow renaturation of polymerase chain reactionproducts allow the formation of heteroduplex DNA that can be detected by itsdifferential mobility in polyacrylamide gel electrophoresis. Heteroduplexanalysis was used to determine if the U1 snRNA microsatellite could be a usefulgenetic marker in Echinococcus granulosus. A U1 snRNA microsatellite fragmentfrom E. granulosus was isolated and characterized by Southern blot andsequencing. Four E. granulosus strains were analyzed: sheep, Tasmanian sheep,cattle, and camel strains. The former two showed polymorphism and shared threeof the six patterns found for sheep strain. The cattle strain displayed twopatterns, and the camel strain was monomorphic. The electrophoretic profileswere used for statistical analysis in order to determine genetic distance andthe relationship among strains. Heteroduplex analysis can be helpful ingenotyping E. granulosus strains and is useful in detecting polymorphism withinstrains.