Bloodcenter Research Foundation

Blood Center Research Foundation, Inc., the

Lee Ann Baxter-Lowe

Jack A Gorski

A method for genetic typing includes the steps of amplifying a genetic sequence of a subject to obtain amplified DNA, which genetic sequence occurs naturally in two or more genetic types, bringing a sample of the amplified DNA into contact with an oligonucleotide probe bound to a support under stringent hybridizing conditions, which oligonucleotide probe hybridizes specifically with DNA having a sequence of one of the genetic types and not with DNA having a sequence of the other genetic types, removing unbound amplified DNA, for example, by washing the support, and analyzing the sample to determine if the one genetic type associated with the probe is present. Use of a solid support such as microbeads provides a more rapid method for identifying polymorphic nucleotide sequences of polymorphic genes, such an HLA sequences.

Method for genetic typing

A method for HLA typing by amplification of a sample followed by sequence-specific oligonucleotide hybridization with a labelled oligonucleotide probe provides for both positive and negative controls. Control sequences representing known allelic polymorphisms at the locus in question are subjected to the labelled probe along with the sample. This method reduces errors and improves the chance of obtaining a successful tissue match, as is vital in the case of tissue transplants, particularly bone marrow transplants. Probes and PCR primers useful in HLA-DR typing are also provided.

Method for HLA typing

The invention provides an improved method for typing specific HLA sequences and other genetic sequences exhibiting similar polymorphism may be detected by sequence specific amplification (SSA). In essence, the primers used to carry out the amplification are chosen to represent sequences that distinguish one allele from another. As a result, the allele is detected if amplification occurs, and is absent if no amplification occurs. This technique can eliminate the need to conduct sequence-specific probe hybridization in order to distinguish among closely similar alleles, and can, in particular, allow resolution of complementary pairs of naturally-occurring HLA alleles which have the same polymorphisms but located on different individual alleles of each pair.

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New Berlin, WI, United States of America

Lee Ann Baxter-Lowe