Journal article
Human platelets as a substrate source for the in vitro amplification of the abnormal prion protein (PrPSc) associated with variant Creutzfeldt-Jakob disease
Transfusion, Vol.49(2), pp.376-384
2009
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.
Details
- Title
- Human platelets as a substrate source for the in vitro amplification of the abnormal prion protein (PrPSc) associated with variant Creutzfeldt-Jakob disease
- Authors/Creators
- M. Jones (Author/Creator) - Western General HospitalA.H. Peden (Author/Creator) - Deanery of Molecular, Genetic and Population Health SciencesH. Yull (Author/Creator)D. Wight (Author/Creator)M.T. Bishop (Author/Creator) - Deanery of Molecular, Genetic and Population Health SciencesC.V. Prowse (Author/Creator)M.L. Turner (Author/Creator) - West Midlands DeaneryJ.W. Ironside (Author/Creator) - Deanery of Molecular, Genetic and Population Health SciencesI.R. MacGregor (Author/Creator)M.W. Head (Author/Creator) - Deanery of Molecular, Genetic and Population Health Sciences
- Publication Details
- Transfusion, Vol.49(2), pp.376-384
- Publisher
- Wiley-Blackwell
- Identifiers
- 991005541311207891
- Copyright
- © 2009 American Association of Blood Banks
- Murdoch Affiliation
- Murdoch University
- Language
- English
- Resource Type
- Journal article
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- Citation topics
- 1 Clinical & Life Sciences
- 1.52 Neurodegenerative Diseases
- 1.52.992 Prion Pathogenesis
- Web Of Science research areas
- Hematology
- ESI research areas
- Clinical Medicine