DETECTION OF 2 POINT MUTATIONS CAUSING FAMILIAL DEFECTIVE APOLIPOPROTEIN-B-100 BY HETERODUPLEX ANALYSIS

Familial defective apolipoprotein B-100 (FDB) is a dominantly-inherite d genetic disorder causing primary hypercholesterolemia and premature coronary heart disease. To date, only two mutations causing FDB have b een identified. A rapid non-radioactive technique is described to dete ct both disease-related apolipoprotein B point mutations in polymerase chain reaction (PCR) products amplified from genomic DNA. Heteroduple x formation between different alleles from FDB heterozygotes was shown to be visible directly after electrophoresis of PCR products and stai ning in low cross-linking polyacrylamide gels. We found that the simpl icity of the method, in addition to its potential to identify new muta tions in the amplified PCR product, makes heteroduplex detection the p referred initial method of screening potential heterozygotes.