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Revista chilena de cirugía

On-line version ISSN 0718-4026

Abstract

MANTEROLA, CARLOS et al. Echinococcus granulosus genotypes in samples of human hydatidosis. Rev Chil Cir [online]. 2006, vol.58, n.6, pp.441-446. ISSN 0718-4026.  http://dx.doi.org/10.4067/S0718-40262006000600008.

Introduction: Environmental variability of Echinococcus granulosus (Eg) has been described. 10 genotypes have been reported as well as a certain intergenotype heterogenicity in studies with material of animal origin. The aim of this study is to describe the results of an Eg genotypification protocol in samples of human hydatidosis. Materials and methods: Cross-sectional study. The hydatid fluid was collected from a consecutive sample of patients operated on for hepatic and pulmonary hydatidosis at hospitals in Temuco between July, 2004 and September, 2005. A DNA extraction protocol was designed for Eg in homogenized samples of hydatid cyst aspirate. 2 polymerase chain reaction (PCR) systems were used: PCREg9 and PCREg16; both in concentrations of 2mM MgCl2. For PCREg9, Eg9F and Eg9R primers were used at concentrations of 0.5 mM, using 35 cycles with a hybridization temperature of 60°C. For PCREg16 Eg16F and Eg16R primers were used at concentrations of 0.5 mM, using 35 cycles with a hybridization temperature of 65°C. The PCR products were absorbed with a restriction enzyme (Rsa1) in order to distinguish the genotypes. Results: 25 samples were analyzed, 4 from pulmonary cysts and 21 from hepatic cysts. Eg amplification was achieved in 22 of the 25 samples (88%). The enzymatic digestion revealed the presence of 3 possible genotypes: in 21 of 22 samples (95.45%) a restriction pattern was observed corresponding to the G1 or G7 genotypes, and the remaining samples to the G4 or G7 genotypes. Conclusion: With the PCR systems used, Eg DNA was detected

Keywords : Echinococcosis"[MeSH]; "Hydatidosis"[Multi]; "Genotype"[MeSH]; "Genotype/classification"[MeSH]; "Polymerase Chain Reaction"[MeSH].

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