Low-resolution DNA typing for HLA-B using sequence-specific primers in allele- or group-specific ARMS/PCR.
The products of the human major histocompatibility complex (HLA Class I and II) have historically been detected using serological or cellular assays. With the availability of DNA sequence information for alleles of the HLA system, and with the development of molecular biological techniques it has become possible to tissue type for allelic differences in the HLA genes themselves. We describe here a polymerase chain reaction (PCR) system, based on the principle of the amplification refractory mutation system (ARMS), for low-resolution DNA typing of the HLA-B gene. The technique involves a one-step PCR from genomic DNA using sequence-specific primers in particular combinations that determine the specificity of each reaction. A low-resolution primer panel has been designed, based on published HLA-B gene nucleotide sequences, consisting of 34 sequence-specific primers (SSP) in 24 PCR reactions which cover all known HLA-B alleles, to give allele-specific or group-specific amplification of DNA fragments of defined size (344-784bp). Advantages of the system are that it can be performed in under 4 hours including DNA extraction, results are easy to interpret and it does not require viable cells.
Tissue Antigen Laboratory, Imperial Cancer Research Fund, London, United Kingdom.
A M
AM
Sadler
F
F
Petronzelli
P
P
Krausa
S G
SG
Marsh
M G
MG
Guttridge
M J
MJ
Browning
J G
JG
Bodmer
D001483:Base Sequence; D004252:DNA Mutational Analysis; D017931:DNA Primers / Q000235:genetics*; D004587:Electrophoresis, Agar Gel; D015235:HLA-B Antigens / Q000235:genetics*; D006801:Humans; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods*; D011110:Polymorphism, Genetic / Q000235:genetics*
D017931:DNA Primers; D015235:HLA-B Antigens