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Identification of Giardia species and Giardia duodenalis assemblages by sequence analysis of the 5.8S rDNA gene and internal transcribed spacers

Published online by Cambridge University Press:  17 March 2010

SIMONE M. CACCIÒ ,
RELJA BECK ,
ANDRE ALMEIDA ,
ANNA BAJER  and
EDOARDO POZIO
SIMONE M. CACCIÒ*
Affiliation:
Department of Infectious, Parasitic and Immunomediated Diseases, Istituto Superiore di Sanità, Rome, Italy
RELJA BECK
Affiliation:
Department of Infectious, Parasitic and Immunomediated Diseases, Istituto Superiore di Sanità, Rome, Italy Department for Bacteriology and Parasitology, Croatian Veterinary Institute, Zagreb, Croatia
ANDRE ALMEIDA
Affiliation:
Department of Infectious, Parasitic and Immunomediated Diseases, Istituto Superiore di Sanità, Rome, Italy Centro de Imunologia e Biologia Parasitária, Instituto Nacional de Saúde Dr Ricardo Jorge, Porto, Portugal
ANNA BAJER
Affiliation:
Department of Parasitology, Faculty of Biology, University of Warsaw, Poland
EDOARDO POZIO
Affiliation:
Department of Infectious, Parasitic and Immunomediated Diseases, Istituto Superiore di Sanità, Rome, Italy
*
*Corresponding author: Department of Infectious, Parasitic and Immunomediated Diseases, Istituto Superiore di Sanità, Viale Regina Elena 299, 00161 Rome, Italy. Tel: +39 06 4990 2484. Fax: +39 06 4990 3561. E-mail: simone.caccio@iss.it
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Summary

PCR assays have been developed mainly to assist investigations into the epidemiology of Giardia duodenalis, the only species in the Giardia genus having zoonotic potential. However, a reliable identification of all species is of practical importance, particularly when water samples and samples from wild animals are investigated. The aim of the present work was to genotype Giardia species and G. duodenalis assemblages using as a target the region spanning the 5.8S gene and the 2 flanking internal transcribed spacers (ITS1 and ITS2) of the ribosomal gene. Primers were designed to match strongly conserved regions in the 3′ end of the small subunit and in the 5′ end of the large subunit ribosomal genes. The corresponding region (about 310 bp) was amplified from 49 isolates of both human and animal origin, representing all G. duodenalis assemblages as well as G. muris and G. microti. Sequence comparison and phylogenetic analysis showed that G. ardeae, G. muris, G. microti as well as the 7 G. duodenalis assemblages can be easily distinguished. Since the major subgroups within the zoonotic assemblages A and B can be identified by sequence analysis, this assay is also informative for molecular epidemiological studies.

Keywords

Giardia duodenalisGiardia murisGiardia microti5.8S rDNAinternal transcribed spacerssequence analysisgenotyping
Type
Research Article
Information
Parasitology , Volume 137 , Issue 6 , May 2010 , pp. 919 - 925
Copyright
Copyright © Cambridge University Press 2010

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