Molecular typing for HLA class I using ARMS‐PCR: Further developments following the 12th International Histocompatibility Workshop

Wiley - Tập 53 Số 2 - Trang 175-183 - 1999
Susan Tonks1, Steven G.E. Marsh2, M. Bunce M3, Walter F. Bodmer4
1Cancer and Immunogenetics Laboratory, Imperial Cancer Research Fund, Institute of Molecular Medicine, John Radcliffe Hospital, Headington, Oxford, UK.
2Anthony Nolan Research Institute, The Royal Free Hospital, Hampstead, London, UK (formerly of the Tissue Antigen Laboratory, Imperial Cancer Research Fund, London, UK)
3Tissue Typing Laboratory, Nuffield Department of Surgery, Oxford Transplant Centre, Churchill Hospital, Oxford, UK
4Cancer and Immunogenetics Laboratory, Imperial Cancer Research Fund, Institute of Molecular Medicine, John Radcliffe Hospital, Headington, Oxford, UK (formerly the Tissue Antigen Laboratory, Imperial Cancer Research Fund, London, UK)

Tóm tắt

Molecular typing for HLA class I was introduced in the 12th International Histocompatibility Workshop. Following a pilot study using three methods, sequence specific oligotyping (SSO), reverse dot blot and amplification refractory mutation system (ARMS)‐PCR, the ARMS‐PCR method was selected for use. A great advantage of an ARMS‐PCR method is that, unlike the other two methods, it can determine whether sequence motifs are in cis or in trans, as ARMS‐PCR detects two cis located motifs per reaction using forward and reverse sequence specific primers. Resolution was designed to be low to medium level for HLA‐A, ‐B and ‐C alleles. Two hundred and fifty class I kits and 83 HLA‐A2 subtyping kits were distributed. The A2 subtyping kit used a two round nested PCR system to identify all of the A2 alleles known at the time. Typing results on control DNA samples distributed with both the kits showed a very satisfactory performance. Since the 12th Workshop, the kits have been developed with the addition of new primers and primer mixes to increase the resolution of the test.

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Wiley

Tập 53 Số 2

175-183

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