Rapid HLA-DQB typing by eight polymerase chain reaction amplifications with sequence-specific primers (PCR-SSP)

M. Bunce; C.J. Taylor; K.I. Welsh

doi:10.1016/0198-8859(93)90502-R

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Rapid HLA-DQB typing by eight polymerase chain reaction amplifications with sequence-specific primers (PCR-SSP)

Journal article

Peer reviewed

Rapid HLA-DQB typing by eight polymerase chain reaction amplifications with sequence-specific primers (PCR-SSP)

M. Bunce, C.J. Taylor and K.I. Welsh

Human Immunology, Vol.37(4), pp.201-206

1993

DOI: https://doi.org/10.1016/0198-8859(93)90502-R

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Abstract

Molecular genotyping of HLA class II genes using group-specific DNA amplification by the PCR followed by probing with (PCR-SSO) probes is too time consuming for the typing of cadaveric organ donors. Recently, amplification of DNA using PCR-SSP has proved a reliable and rapid method for typing HLA-DRB1 genes. PCR-SSP takes 2 hours to perform and is therefore suitable for the genotyping of cadaveric donors. We have designed a set of primers that in eight PCR reactions will positively identify the HLA-DQB1 alleles corresponding o the serologically defined series HLA-DQ2, DQ4, DQ5, DQ6, DQ7, DQ8, and DQ9. Presently, 30 homozygous cell lines and 138 individuals have been typed by the DQB1 PCR-SSP technique and compared with a combination of serology and RFLP with 100% concordance. No false-negative or false-positive amplifications were recorded. All combinations of DQB1 can be readily identified. DQB1 PCR-SSP can take as little as 130 minutes from start to finish, including DNA preparation.

Details

Title: Rapid HLA-DQB typing by eight polymerase chain reaction amplifications with sequence-specific primers (PCR-SSP)
Authors/Creators: M. Bunce (Author/Creator) - Churchill Hospital
C.J. Taylor (Author/Creator) - Churchill Hospital
K.I. Welsh (Author/Creator) - Churchill Hospital
Publication Details: Human Immunology, Vol.37(4), pp.201-206
Publisher: Elsevier BV
Identifiers: 991005540151507891
Copyright: 1993 Elsevier Inc.
Murdoch Affiliation: Murdoch University
Language: English
Resource Type: Journal article

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Citation topics: 1 Clinical & Life Sciences; 1.6 Immunology; 1.6.607 MHC Diversity
Web Of Science research areas: Immunology
ESI research areas: Immunology