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Human platelets as a substrate source for the in vitro amplification of the abnormal prion protein (PrPSc) associated with variant Creutzfeldt-Jakob disease

Jones, M.ORCID: 0000-0001-5002-0227, Peden, A.H., Yull, H., Wight, D., Bishop, M.T., Prowse, C.V., Turner, M.L., Ironside, J.W., MacGregor, I.R. and Head, M.W. (2009) Human platelets as a substrate source for the in vitro amplification of the abnormal prion protein (PrPSc) associated with variant Creutzfeldt-Jakob disease. Transfusion, 49 (2). pp. 376-384.

Link to Published Version: https://doi.org/10.1111/j.1537-2995.2008.01954.x
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Abstract

BACKGROUND: Four recent cases of transfusion‐related transmission of variant Creutzfeldt‐Jakob disease (vCJD) highlight the need to develop a highly sensitive and specific screening test to detect infectivity in the blood of asymptomatic infected individuals. Protein misfolding cyclic amplification (PMCA), a method for the amplification of minute amounts of disease‐associated abnormal prion protein (PrPSc) to readily detectable levels, could be incorporated into such a test provided that a suitable substrate source for routine use in human PMCA reactions can be found.

STUDY DESIGN AND METHODS: With the use of seed sources from individuals with variant and sporadic CJD, the use of human platelets (PLTs) as a PMCA substrate source was evaluated. The effects of seed/substrate prion protein gene (PRNP) codon 129 genotype compatibility on amplification efficiency and freeze‐thaw on a substrate's ability to support amplification and the degree of amplification achieved by serial PMCA (sPMCA) were investigated.

RESULTS: Seed/substrate PRNP codon 129 compatibility was found to have a major influence on PrPSc amplification efficiency. Individual substrates, of the same PRNP codon 129 genotype, could be pooled and stored frozen for use in subsequent PMCA reactions. A consistent 10‐fold increase in PrPSc detection sensitivity was achieved after each round of sPMCA, resulting in a 10,000‐fold increase in detection sensitivity after four rounds, with no evidence of de novo PrPSc production detected in the unseeded PLT substrate.

CONCLUSIONS: Providing issues of seed/substrate PRNP codon 129 compatibility are taken into consideration human PLTs are a suitable, readily available, renewable substrate source for use in human PMCA applications.

Item Type: Journal Article
Publisher: Wiley-Blackwell
Copyright: © 2009 American Association of Blood Banks
URI: http://researchrepository.murdoch.edu.au/id/eprint/49451
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