RAPID, EFFICIENT METHOD FOR MULTIPLEX AMPLIFICATION FROM FILTER-PAPER

Guthrie cards derived from the New York State Newborn Screening Progra m were utilized to develop a rapid, economical method for amplifying m ultiple genes to detect mutations that impact public health. These spe cimens are untraceable to the donor because identifiers are removed an d discarded; there fore, these pilot studies were carried out anonymou sly. The sample preparation requires minimal ma nipulation, is amenabl e to automation, and is useful in laboratories which routinely process large numbers of samples, such as those in typical newborn screening laboratories. Multiple gene fragments may be amplified from a 1 mm pun ch which contains less than 1 mu l of whale blood. The blood spots use d in these studies contain sufficient material for up to 25 amplificat ion reactions which multiplex at least four different gene fragments e ach. Since sufficient material remains on the card after the routine t esting is complete, this simple assay can greatly expand the efficacy of current newborn screening programs by permitting DNA diagnosis of s ome disorders when indicated, particularly those in which genotype-phe notype correlations are useful. In addition to newborn screening speci mens, this method is also applicable to whole blood from adults after phlebotomy and from lymphoblastoid cell lines. Use of filter paper for DNA analysis is particularly useful for shipped specimens or for popu lation studies whose subjects are refractory to phlebotomy. (C) 1995 W iley Liss, Inc.