DNA-BASED HLA CLASS-II POSTMORTEM TYPING - EVALUATION OF DIFFERENT TECHNIQUES FOR PROSPECTIVE CORNEAL ALLOGRAFTING

Background: The objective of this study was to establish DNA-based HLA -DR postmortem tissue typing techniques in order to improve the qualit y and quanity of fully HLA-typed corneas for prospective allografting. Methods: Four hundred and thirty-seven cornea donors were investigate d. DNA was derived from cultivated retinal pigment epithelial cells by spin column purification or a salting out technique, and from scleral tissue by a very simple boiling method. Donors were typed by hybridiz ation of polymerase chain reaction (PCR) products with sequence-specif ic oligodesoxynucleotide probes (PCR-SSOP) or by PCR with sequence-spe cific primers (PCR-SSP). Twenty-two of the donors were pretyped by ser ology. Results: We observed high concordance (96%) between the results of DNA-based postmortem typing and the serological lymphocytotoxicity test. Furthermore, the distribution of the HLA-DR specificities that were detected correlated well with the distribution in a control popul ation. Conclusions: The results of this study demonstrate that both PC R SSP and PCR-SSOP allow prospective allocation of HLA class II-matche d corneas with high accuracy.