NR ALQE
AU Teupser,D.; Heino,N.; Wilfert,W.; Thiery,J.
TI Rapid detection of the prion protein M129V polymorphism with the LightCycler
QU Journal of Neuroscience Methods 2002 Mar 30; 115(1): 93-6
PT journal article
AB The common single nucleotide polymorphism at codon 129 of the prion protein gene is a key determinant of the genetic susceptibility to Creutzfeldt-Jakob disease (CJD). Recently, a molecular classification of sporadic CJD based on the M129V genotype in conjunction with other determinants was proposed. In the present study, we describe the development and evaluation of a rapid fluorescent-based assay to detect this polymorphism using the LightCycler system. The two polymorphic alleles could be clearly distinguished by their melting points at 52.1 and 60.4 degrees C, representing the 129V and 129M alleles, respectively. These results were confirmed by DNA sequencing. We evaluated our test in 400 patient samples and found no deviations from the expected melting patterns. The calculated allele frequency for the M-allele was 0.66. Thus, we have established a rapid, reliable fluorescent assay for high-throughput detection of the prion protein M129V polymorphism.
MH Creutzfeldt-Jakob Syndrome/blood/*diagnosis/*genetics; DNA Mutational Analysis/instrumentation/methods; Fluorescent Dyes/*diagnostic use; Genetic Screening/instrumentation/*methods; Genetic Techniques/instrumentation; Genotype; Human; Mutation/*genetics; Photic Stimulation; Polymerase Chain Reaction/instrumentation/methods; Polymorphism (Genetics)/*genetics; Prions/blood/*genetics
AD Institute of Laboratory Medicine, Clinical Chemistry and Molecular Diagnostics, University Hospital Leipzig, Liebigstr. 27, 04103 Leipzig, Germany. daniel@teupser.de
SP englisch
PO Niederlande