DELETION DETECTION IN THE DYSTROPHIN GENE BY MULTIPLEX GAP LIGASE CHAIN-REACTION AND IMMUNOCHROMATOGRAPHIC STRIP TECHNOLOGY

The purpose of this study is to demonstrate the value of a multiplex a mplification and readout system. The validation was done using as a mo del system the detection of deletions in nine possible dystrophin exon s: 4, 8, 12, 17, 19, 44, 45, 48, and 51. The amplification system was gap ligase chain reaction, adapted to amplify selected regions of mult iple exons simultaneously. The amplified products were read out with a n immunochromatographic methodology, adapted from that used in the Abb ott product line commercialized under the name Test Pack Plus. In each amplification, the beta-globin gene was incorporated and served as a procedural control. The complete process takes <3 hr from DNA sample t o result. The procedure is therefore rapid and simple, as well as bein g potentially very cost effective. The combination of these two techno logies is shown to be a useful tool for the determination of deletions in the nine exons of the dystrophin gene. The results of a 100-patien t sample study showed concordance with cDNA and PCR in current use. Eq uivalent performance at two sites was shown. (C) 1995 Wiley Liss, Inc.