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Apolipoprotein E genotyping in patients with neurodegenerative diseases
L A Ballering1, H M Steffens-Nakken, R A Esselink
1Department of Clinical Chemistry, Medical Spectrum Twente, Hospital Group, Enschede, The Netherlands.
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 semi-nested PCR followed by hybridization with biotin-labeled allele-specific 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. Seventy-six 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.