A rapid and efficient method for the detection of point mutations of the human prion protein gene (PRNP) by direct sequencing

Creutzfeldt-Jakob disease (CJD) and related disorders occur in sporadic, ac quired and inherited forms. In sporadic, iatrogenic and new variant CJD the polymorphic codon 129 of the prion protein gene (PRNP) plays an important role for the susceptibility to the disease and for the clinical and neuropa thological manifestations. All the inherited forms of CJD and related disor ders are linked to point or insert mutations of PRNP. The analysis of PRNP is therefore important for a correct classification of these disorders and for the identification of novel mutations. The aim of the present study is to describe a fast and easy to perform method for the direct sequencing of the PCR amplified PRNP open reading frame, by using M13 tailed primers whic h allow a direct and rapid method of sequencing. The goodness of this metho d is demonstrated in the analysis of three sporadic CJD patients with diffe rent genotypes at codon 129 and three inherited cases bearing different poi nt mutations of PRNP: the Pro102Leu mutation linked to Gerstmann - Straussl er - Scheinker-syndrome, the Val210Ile mutation and a novel mutation at cod on 211 (Gln211Glu) both associated to familial CJD. (C) 2000 Elsevier Scien ce B.V. All rights reserved.