Journal
TRANSFUSION
Volume 49, Issue 2, Pages 376-384Publisher
WILEY-BLACKWELL
DOI: 10.1111/j.1537-2995.2008.01954.x
Keywords
-
Categories
Funding
- Medical Research Council [G0600953, G0700711B, G0900580] Funding Source: researchfish
- Medical Research Council [G0900580, G0600953] Funding Source: Medline
- MRC [G0600953, G0900580] Funding Source: UKRI
Ask authors/readers for more resources
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. 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. 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. 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.
Authors
I am an author on this paper
Click your name to claim this paper and add it to your profile.
Reviews
Recommended
No Data Available