APOLIPOPROTEIN-E GENOTYPING IN PATIENTS WITH NEURODEGENERATIVE DISEASES

Objectives: There is a growing demand to perform apolipoprotein E (Apo -E) genotyping on neuropathologic archive material. Due to the extreme ly long fixation time, this material is unsuitable for routinely used Apo-E genotyping methods. We present an investigation into the applica bility of a new method. Design and Methods: An Apo-E genotyping method was tested for use on formalin-fixed paraffin-embedded brain tissue, using semi-nested PCR followed by hybridization with biotin-labeled al lele-specific oligonucleotides, and chemiluminescent detection. The me thod was applied to 88 archive samples of different neurologic disorde rs. Results: With this technique 76% (67/88) of the samples could be g enotyped. The crucial step is the semi-nested PCR. All the samples fro m which a PCR product could be obtained, the Apo-E gene could be genot yped without interpretation problems. Seventy-six percent of the sampl es that could not be genotyped, were fixed in unbuffered formalin. Con clusions: This technique offers a good Apo-E genotyping method applica ble on neuropathological archive material in order to support in retro spect clinical studies.