Method and kit for amplification, sequencing and typing of class

Details Method and kit for amplification, sequencing and typing of class Method and kit for amplification, sequencing and typing of class

1997-10-10

1999-06-08

Saunders, David

Chemistry: molecular biology and microbiology

Measuring or testing process involving enzymes or...

Involving nucleic acid

435810, 536 2431, 536 2433, C12Q 168, C07H 2104

Patent

active

059104139

ABSTRACT:
Amplification primer and sequencing primer sites have been identified which permit sequence-based typing of each of the classical HLA genes in highly robust and consistent reactions without allelic dropout. To determine the DNA sequence, and thus the type of at least one exon of an HLA-A, HLA-B and HLA-C gene present in a sample, the sample is combined with an amplification reaction mixture containing the amplification primers and amplified to form an amplification product including exon 2 and exon 3 of the gene together in a single fragment. The amplification product is then combined with a sequencing reaction mixture containing one or more oligonucleotide sequencing primers which hybridize to a conserved regions the amplification product. The oligonucleotide sequencing primers between them are effective to produce sequencing fragments from all known alleles of exon 2 or exon 3 of the gene under suitable conditions to produce sequencing fragments. The sequencing fragments are then evaluated to determine the sequence of exon 2 or exon 3 of the gene in the sample.

REFERENCES:
patent: 5424184 (1995-06-01), Santamaria et al.
patent: 5451512 (1995-09-01), Apple et al.
Johnstoan-Dow, et al. "A General Approach for Sequencing Based Typing of HLA-A" Poster Presentation, 1995 ASHI Meeting, Dallas, TX.
Blasczyk et al., "The Diversoty of the HLA Class I Introns Reflects the Ancestral Relationships of the Coding Regions", Abstracts of the 22.sup.nd Annual Meeting if The American Society of Histocompatability and Immunogenetics, No. 2.4-26 (1996).
Cereb et al., Tissue Antigens 45: 1-11 (1995).
Weiss et al., "Organization, Sequence and expression of the HLA-B27 Gene: Moelcular Approach to Analyze HLA and Disease Associations"Immunobiol. 170: 367-380 (1985).
Blascyzk et al., Tissue Antigens 47: 102-110 (1996).
Petersdorf et al., "A comprehensive approach for typing the alleles of the HLA-B locus by automated Sequencing" Tissue Antigens 46: 73-85 (1995).
Santamaria et al., Human Immunology 37: 39-50 (1993).
Saiki, et al., Proc. Natl Acad. Sci. (USA) 86: 6230-6234 (1989).
"HLA Class I SSP ARMS-PCR Typing Kit", Reference Manual (1995).

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