Genotyping Cryptosporidium parvum by single-strand conformation polymorphism analysis of ribosomal and heat shock gene regions

R.B. Gasser; X-Q Zhu; S.M. Cacciò; R.M. Chalmers; G. Widmer; U.M. Morgan; R.C.A. Thompson; E. Pozio; G.F. Browning

doi:10.1002/1522-2683(200102)22:3<433::AID-ELPS433>3.0.CO;2-K

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Genotyping Cryptosporidium parvum by single-strand conformation polymorphism analysis of ribosomal and heat shock gene regions

Journal article

Peer reviewed

Genotyping Cryptosporidium parvum by single-strand conformation polymorphism analysis of ribosomal and heat shock gene regions

R.B. Gasser, X-Q Zhu, S.M. Cacciò, R.M. Chalmers, G. Widmer, U.M. Morgan, R.C.A. Thompson, E. Pozio and G.F. Browning

Electrophoresis, Vol.22(3), pp.433-437

2001

DOI: https://doi.org/10.1002/1522-2683(200102)22:3<433::AID-ELPS433>3.0.CO;2-K

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Abstract

Polymerase chain reaction (PCR)-coupled single-strand conformation polymorphism (SSCP) approaches utilizing nuclear DNA regions of the small subunit (SSU) of ribosomal RNA and heat shock protein 70 gene (HSP70) were established for genotyping Cryptosporidium parvum. The regions were amplified (individually or in a multiplex reaction) by PCR from DNA extracted from oocysts from ruminant or human hosts, then denatured and subjected to electrophoresis in a mutation detection enhancement (nondenaturing) gel matrix. Single-strand profiles produced in SSCP allowed the unequivocal identification/differentiation of the two common (human, 1 and cattle, 2) genotypes of C. parvum and the direct display of sequence variability within some samples, reflecting population variation. As these are considered among the most closely related genotypes (based on SSU and HSP70 sequence data), these findings and other preliminary results for C. felis (from cat) C. serpentis (from snake) and C. baileyi (from bird) indicate that the SSCP approaches established could be employed to identify any of the currently recognised genotypes and species of Cryptosporidium.

Details

Title: Genotyping Cryptosporidium parvum by single-strand conformation polymorphism analysis of ribosomal and heat shock gene regions
Authors/Creators: R.B. Gasser (Author/Creator) - The University of Melbourne
X-Q Zhu (Author/Creator) - The University of Melbourne
S.M. Cacciò (Author/Creator) - Istituto Superiore di Sanità
R.M. Chalmers (Author/Creator) - Singleton Hospital
G. Widmer (Author/Creator) - Tufts University
U.M. Morgan (Author/Creator) - World Health Organization - Pakistan
R.C.A. Thompson (Author/Creator) - World Health Organization - Pakistan
E. Pozio (Author/Creator) - Istituto Superiore di Sanità
G.F. Browning (Author/Creator) - The University of Melbourne
Publication Details: Electrophoresis, Vol.22(3), pp.433-437
Publisher: John Wiley & Sons Ltd
Identifiers: 991005542297007891
Copyright: © 2001 Wiley-VCH Verlag GmbH
Murdoch Affiliation: School of Veterinary and Biomedical Sciences
Language: English
Resource Type: Journal article

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Collaboration types: Domestic collaboration; International collaboration
Citation topics: 1 Clinical & Life Sciences; 1.246 Diarrheal Diseases; 1.246.985 Cryptosporidium
Web Of Science research areas: Biochemical Research Methods; Chemistry, Analytical
ESI research areas: Chemistry