Elettroforesi zonale capillare: Una nuova tecnica per la diagnosi molecolare di malattie genetiche

Capillary zone electrophoresis (CZE) in polymer networks was applied to DNA separation. Conditions were set up to the analysis of PCR products in the 50-500 base pairs (bp) range in capillaries filled with low viscosity liquid linear polyacrylamide. CZE was applied to the diagnosis of different genetic diseases: Cystic Fibrosis (CF), Duchenne (DMD)/Becker (BMD) Muscular Dystrophy, 21-Hydroxylase Deficiency and Androgen Insensitivity Syndrome. A number of mutations and polymorphisms, including deletions, point mutations, point mutations altering restriction sites and variable number of tandem repeats (VNTR), have been analyzed by this technique, in parallel with standard electrophoresis procedures. Detection systems for multiple amplification reactions for deletion identification in DMD/BMD, and for mutation analysis in CF, have been developed. CZE in temporal thermal gradients (TGCE) was developed, in alternative to Denaturing Gradient Gel Electrophoresis (DGGE). The temperature gradient was generated internally by using a dedicated computer program able to calculate the precise inner temperature under given electric conditions. This method was applied to the identification of point mutations in CF. Due to the ease of operation and multi-sample load ability, CZE could represent a valid alternative over existing techniques.