HLA typing by Reference Strand Mediated Conformation Analysis (RSCA).
Reference Strand Mediated Conformation Analysis (RSCA) is a novel conformational method that offers high resolution and high sample throughput for typing the HLA class I and class II genes. This technique differs from conventional sequence-based typing methodologies in that the HLA type is assigned on the basis of accurate measurement of conformation-dependent DNA mobility in polyacrylamide gel electrophoresis (AGE). RSCA utilises a fluorescent-labelled locus-specific reference DNA to selectively modify the molecular conformation of the tested DNA. The use of laser-based instrumentation and computer software, in addition to internal DNA markers for correction of gel variability, allows the discrimination of HLA alleles which differ by one nucleotide in a DNA fragment nearly as large as a kilobase in length. RSCA has been successfully applied in blind studies of HLA typing, demonstrating that is reproducible, able to identify new alleles and able to resolve ambiguous heterozygous combinations.
Anthony Nolan Research Institute, The Royal Free Hospital, London, UK.
J R
JR
Argüello
J A
JA
Madrigal
D004247:DNA / Q000032:analysis; D005456:Fluorescent Dyes; D006680:HLA Antigens / Q000235:genetics*; D006650:Histocompatibility Testing / Q000379:methods*; D006801:Humans; D009690:Nucleic Acid Conformation
D005456:Fluorescent Dyes; D006680:HLA Antigens; D004247:DNA