HOW SENSITIVE IS PCR-SSCP

Single-strand conformation polymorphism analysis (SSCP) is a rapid met hod for detection of minor sequence changes in polymerase chain reacti on-amplified DNA. Since the first reported use of SSCP in 1989 (Orita et al., 1989), this tehnique has been used widely to detect mutations in oncogenes, tumor suppressor genes, and genes responsible for geneti c diseases. Published mutations that have been detected using this tec hnique include base substitutions, small insertions and deletions, and rearrangements. This technique has also been applied for the detectio n of DNA polymorphisms at various loci of the human genome (reviewed b y Hayashi, 1991; Hayashi, 1993). However, many factors can influence t he sensitivity of SSCP, and its optimization is highly empirical. In t his review, we estimate the percentage of mutations that can be detect ed by this technique under various controlled conditions, and describe some critical elements affecting sensitivity. (C) 1993 Wiley-Liss, In c.