Whole Genome Amplification (WGA) for archiving and genotyping of clinical isolates of Cryptosporidium species

Bouzid, Maha, Heavens, Darren, Elwin, Kristin, Chalmers, Rachel M., Hadfield, Stephen J., Hunter, Paul R. ORCID: https://orcid.org/0000-0002-5608-6144 and Tyler, Kevin M. ORCID: https://orcid.org/0000-0002-0647-8158 (2010) Whole Genome Amplification (WGA) for archiving and genotyping of clinical isolates of Cryptosporidium species. Parasitology, 137 (1). pp. 27-36. ISSN 1469-8161

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Abstract

Clinical and environmental isolates of pathogens are often unique and may be unculturable, yielding a very limited amount of DNA for genetic studies. Cryptosporidium in particular are difficult to propagate. Whole genome amplification (WGA) is a valuable technique for amplifying genomic material. In this study, we tested 5 WGA commercial kits using Cryptosporidium clinical isolates. DNA of 5 C. hominis and 5 C. parvum clinical isolates and C. parvum IOWA reference strain were used. The majority of the samples were amplified by all of the kits tested. The integrity and fidelity of the amplified genomic DNA were assessed by sequence analysis of several PCR products of varying length. We found evidence that one kit in particular may be more error prone while another seemed the more suitable kit for Cryptosporidium clinical samples, generating high molecular weight DNA from all the samples with high fidelity. Thus WGA was found to be a useful technique for producing amplified DNA suitable for downstream genotyping techniques and archiving of Cryptosporidium clinical isolates.

Item Type: Article
Faculty \ School:
Faculty of Medicine and Health Sciences > Norwich Medical School
University of East Anglia Research Groups/Centres > Theme - ClimateUEA
UEA Research Groups: Faculty of Medicine and Health Sciences > Research Groups > Public Health and Health Services Research (former - to 2023)
Faculty of Medicine and Health Sciences > Research Groups > Epidemiology and Public Health
Faculty of Social Sciences > Research Centres > Water Security Research Centre
Faculty of Medicine and Health Sciences > Research Centres > Population Health
Faculty of Medicine and Health Sciences > Research Centres > Metabolic Health
Depositing User: EPrints Services
Date Deposited: 25 Nov 2010 11:12
Last Modified: 19 Oct 2023 00:34
URI: https://ueaeprints.uea.ac.uk/id/eprint/14953
DOI: 10.1017/S0031182009991132

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