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HLA-B typing by reference strand mediated conformation analysis using a capillary-based semiautomated genetic

D Turner¹, S Akpe, J Brown

¹Department of Histocompatibility and Immunogenetics, North London Centre, National Blood Service, London, UK. david.turner@nbs.nhs.uk

Human Immunology

|April 11, 2001

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Capillary Reference Strand Conformation Analysis (RSCA) is a new method for Human Leukocyte Antigen B (HLA-B) typing. This technique shows promise for accurate and efficient HLA-B allele identification in clinical settings.

Area of Science:

Immunogenetics
Molecular Biology
Human Leukocyte Antigen (HLA) Typing

Background:

Reference Strand Conformation Analysis (RSCA) has been applied to HLA-A typing using capillary electrophoresis.
There is a need for efficient and accurate methods for Human Leukocyte Antigen B (HLA-B) typing.

Purpose of the Study:

To develop and validate capillary RSCA for HLA-B typing.
To define mobility values for HLA-B alleles using capillary electrophoresis.

Main Methods:

Capillary electrophoresis was used to analyze 93 HLA-B alleles.
Simultaneous electrophoresis of three fluorescently labelled references with different dyes.
Validation by comparing capillary RSCA results with reverse SSO typing in 296 cord blood donors.

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Main Results:

Mobility values for 93 HLA-B alleles were established.
14.5% of samples required confirmatory typing after capillary RSCA, versus 5.1% for reverse SSO.
100% correlation was observed between methods when no further typing was needed.

Conclusions:

Capillary RSCA is a quick and easily implementable method for HLA-B typing.
The technique has potential for high-resolution typing with advancements in fluorescent labels and gel matrices.