polipoprotein 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 extremely long fixation time, this material is unsuitable for routinely used Apo-E genotyping methods. We present an investigation into the applicability of a new method. Design and Methods: An Apo-E genotyping method was tested for use on formalin-fixed paraffin-embedded brain tissue, using seminested PCR followed by hybridization with biotin-labeled allelespecific oligonucleotides, and chemiluminescent detection. The method was applied to 88 archive samples of different neurologic disorders. Results: With this technique 76% (67/88) of the samples could be genotyped. The crucial step is the semi-nested PCR. All the samples from which a PCR product could be obtained, the Apo-E gene could be genotyped without interpretation problems. Seventysix percent of the samples that could not be genotyped, were fixed in unbuffered formalin. Conclusions: This technique offers a good Apo-E genotyping method applicable on neuropathological archive material in order to support in retrospect clinical studies.