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Sequence and PCR–RFLP analysis of the internal transcribed spacers of the rDNA repeat unit in isolates of Cryptosporidium from different hosts

Published online by Cambridge University Press:  01 January 1999

U. M. MORGAN ,
P. DEPLAZES ,
D. A. FORBES ,
F. SPANO ,
H. HERTZBERG ,
K. D. SARGENT ,
A. ELLIOT  and
R. C. A. THOMPSON
U. M. MORGAN
Affiliation:
World Health Organization Collaborating Centre for the Molecular Epidemiology of Parasitic Infections and State Agricultural Biotechnology Centre, Division of Veterinary and Biomedical Sciences, Murdoch University, Murdoch, WA 6150, Australia
P. DEPLAZES
Affiliation:
Institute of Parasitology, University of Zurich, Wintherthurerstrasse 266a, CH-8057, Zurich, Switzerland
D. A. FORBES
Affiliation:
Department of Paediatrics, University of Western Australia, Princess Margaret Hospital, GPO Box D184, WA 6001, Australia
F. SPANO
Affiliation:
Istituto di Parassitologia, Università di Roma ‘La Sapienza’, P. le A. Moro 5, 00185 Rome, Italy
H. HERTZBERG
Affiliation:
Institute of Parasitology, University of Zurich, Wintherthurerstrasse 266a, CH-8057, Zurich, Switzerland
K. D. SARGENT
Affiliation:
World Health Organization Collaborating Centre for the Molecular Epidemiology of Parasitic Infections and State Agricultural Biotechnology Centre, Division of Veterinary and Biomedical Sciences, Murdoch University, Murdoch, WA 6150, Australia
A. ELLIOT
Affiliation:
World Health Organization Collaborating Centre for the Molecular Epidemiology of Parasitic Infections and State Agricultural Biotechnology Centre, Division of Veterinary and Biomedical Sciences, Murdoch University, Murdoch, WA 6150, Australia
R. C. A. THOMPSON
Affiliation:
World Health Organization Collaborating Centre for the Molecular Epidemiology of Parasitic Infections and State Agricultural Biotechnology Centre, Division of Veterinary and Biomedical Sciences, Murdoch University, Murdoch, WA 6150, Australia
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Abstract

The Cryptosporidium ITS1, 5·8S and ITS2 rDNA regions from a number of Cryptosporidium isolates from different hosts and geographical areas were cloned and sequenced in order to investigate the extent of sequence heterogeneity between human and cattle-derived isolates from different geographical locations and also between isolates of Cryptosporidium from different hosts such as cats, pigs, mice and a koala. Calf-derived isolates from different continents were virtually identical as were human-derived isolates from the UK and Australia. Genetic differences between Cryptosporidium isolates were extensive and were in fact greater than the level of nucleotide divergence between Toxoplasma gondii and Neospora caninum rDNA sequences. Based on the sequence information derived from this study, PCR–RFLP of the ITS1 region was undertaken in order to directly amplify and genotype Cryptosporidium isolates from different hosts. This PCR–RFLP approach can now be used for molecular epidemiology studies, circumventing the need for costly sequencing and allowing a wider range of genetically different isolates to be examined.

Keywords

CryptosporidiumrDNAITSgenotypes
Type
Research Article
Information
Parasitology , Volume 118 , Issue 1 , January 1999 , pp. 49 - 58
Copyright
1999 Cambridge University Press

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