MOLECULAR SCREENING OF GENETIC-DEFECTS WITH RNA-SSCP ANALYSIS - THE PKU AND CYSTINURIA MODEL

RNA single-strand conformation polymorphism (rSSCP) is a recently deve loped method for detecting genetic defects. This technique requires DN A amplification with a polymerase chain reaction making use of one T7 promoter-containing primer. Amplification products are subsequently tr anscribed in vitro and the labelled transcripts are analysed for singl e-strand conformation changes. rSSCP has been applied to mutation scre ening of the phenylalanine hydroxylase gene and rBAT cDNA, from PKU an d cystinuric patients, respectively. Experimental evidence shows that 83% and 86% of screened PKU and cystinuric mutations, respectively, gi ve rise to detectable rSSCP signals. Thus, results obtained show that RNA single-strand conformation polymorphism analysis is generally appl icable and is a suitable technique for detecting genetic disease causi ng mutations, both in basic research and in clinical practice.