APPLICATIONS OF CONSENSUS POLYMERASE CHAIN-REACTION WITH SUBSEQUENT ELECTROPHORETIC DISTINCTION OF AMPLIFICATES

Conserved sequences within gene families permit the design of consensu s primers that match several members of a given class of homologous ge nes. Polymerase chain reaction (PCR) products obtained with such conse nsus primers were characterized by restriction mapping or single-stran d conformation polymorphism (SSCP) analysis, using precast polyacrylam ide minigels and automated silver staining. Examples for the electroph oretic distinction of consensus amplificates are presented in the fiel ds of guanylyl cyclase expression studies and in the determination of B-cell clonality in human blood samples. Guanylyl cyclase expression i n inner ear tissues of guinea pigs was investigated by reverse transcr iption PCR using consensus primers with specificity for the subclass o f particulate guanylyl cyclases. nle resulting PCR products were assig ned to three representatives of this group by restriction mapping. The consensus PCR approach enabled the detection of an unexpected recepto r type, namely guanylyl cyclase C, in the inner ear, The distinction b y SSCP analysis of denatured consensus amplificates was appropriate fo r the identification of clone-specifically rearranged immunoglobulin h eavy chain genes of B-lymphocytes. Genomic DNA isolated from blood sam ples of leukemia patients served as the template for the consensus amp lification of clone-specific VDJ rearrangements, Rapid distinction and re-identification of consensus PCR products was achieved by SSCP anal ysis for regular antigen receptor rearrangements and for t(14; 18) tra nslocations, The potential of these procedures for detecting leukemia or lymphoma clones when monitoring minimal residual disease was assess ed.