Abstract
HLA typing by sequence-specific primers (PCR-SSP) is a commonly used technique in HLA typing in which multiple pairs of cis-located allele-specific primers are used to determine the alleles present in a given DNA sample. Although the technique is around two decades old, it still offers a relatively straight forward way of typing and has the benefit of using commonly available laboratory equipment. Here we describe the background of the PCR-SSP, how to design and validate reactions, and common problems that arise.
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References
Tindall KR, Kunkel TA (1988) Fidelity of DNA synthesis by the Thermus aquaticus DNA polymerase. Biochemistry 27:6008–6013
Wu DY, Ugozzoli L, Pal BK, Wallace RB (1989) Allele-specific enzymatic amplification of beta-globin genomic DNA for diagnosis of sickle cell anaemia. Proc Natl Acad Sci USA 86:2757–2760
Newton CR, Graham A, Heptinstall LE, Powell SJ, Summers C et al (1989) Analysis of any point mutation in DNA. The amplification refractory mutation system (ARMS). Nucleic Acid Res 17:2503–2516
Fugger L, Morling N, Ryder LP, Odum N, Svejgaard A (1990) Technical aspects of typing for HLA-DP alleles using allele-specific DNA in vitro amplification and sequence-specific oligonucleotide probes. Detection of single base mismatches. J Immunol Methods 129:175–185
Olerup O, Zetterquist H (1992) HLA-DR typing by PCR amplification with sequence-specific primers (PCR-SSP) in 2 hours: an alternative to serological DR typing in clinical practice including donor-recipient matching in cadaveric transplantation. Tissue Antigens 39:225–235
Browning MJ, Krausa P, Rowan A, Bicknell DC, Bodmer JG et al (1993) Tissue typing the HLA-A locus from genomic DNA by sequence-specific PCR: comparison of HLA genotype and surface expression on colorectal tumour cell lines. Proc Natl Acad Sci USA 90:2842–2845
Bunce M, O’Neill CM, Barnardo MC, Krausa P, Browning MJ et al (1995) Phototyping: comprehensive DNA typing for HLA-A, B, C, DRB1, DRB3, DRB4, DRB5 and DQB1 by PCR with 144 primer mixes utilising sequence-specific primers (PCR-SSP). Tissue Antigens 46
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Bunce, M., Passey, B. (2013). HLA Typing by Sequence-Specific Primers. In: Zachary, A., Leffell, M. (eds) Transplantation Immunology. Methods in Molecular Biology, vol 1034. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-493-7_8
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DOI: https://doi.org/10.1007/978-1-62703-493-7_8
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Publisher Name: Humana Press, Totowa, NJ
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