| 摘要 |
<p>A method for the identification of DNA sequence elements in complex and high ly variable sequences is described. The method consists of identifying a short sequence element of several DNA bases (2-6 bases) at a given position in the genome simultaneously on all parental alleles. The method allows differentiating mini-haplotypes on different alleles in one analysis. The method consists of carrying out an enzymatic primer extension reaction with a combination of extension primers (pool of primers) and analysing the product s by mass spectrometry. The pool of primers is assembled in such a way that th e primer extension product allows unambiguous identification of both the prime r of the pool that was extended and the base that was added. The method is of great utility for DNA sequences harbouring many SNPs close to each other wit h many possible haplotypes. Such sequences are known in the Major Histocompatibility Complex (MHC). This method is particularly well suited fo r DNA-based HLA typing and in combination with a suitable selection of sites tested, it is superior in ease of operation to conventional HLA typing methods. We have identified sets of these assays for HLA-A, HLA-B, and HLA-D RB 1 that allow unambiguous four-digit HLA of each of these genes with between 11 and 28 queried markers.</p> |
