Jove
Visualize
Contact Us
JoVE
x logofacebook logolinkedin logoyoutube logo
ABOUT JoVE
OverviewLeadershipBlogJoVE Help Center
AUTHORS
Publishing ProcessEditorial BoardScope & PoliciesPeer ReviewFAQSubmit
LIBRARIANS
TestimonialsSubscriptionsAccessResourcesLibrary Advisory BoardFAQ
RESEARCH
JoVE JournalMethods CollectionsJoVE Encyclopedia of ExperimentsArchive
EDUCATION
JoVE CoreJoVE BusinessJoVE Science EducationJoVE Lab ManualFaculty Resource CenterFaculty Site
Terms & Conditions of Use
Privacy Policy
Policies

Related Experiment Videos

HLA typing by Reference Strand Mediated Conformation Analysis (RSCA).

J R Argüello1, J A Madrigal

  • 1Anthony Nolan Research Institute, The Royal Free Hospital, London, UK.

Reviews in Immunogenetics
|March 20, 2001
PubMed
Summary
This summary is machine-generated.

Related Concept Videos

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by
Same author

High-resolution HLA allele and haplotype frequencies in majority and minority populations of Costa Rica and Nicaragua: Differential admixture proportions in neighboring countries.

HLA·2018
Same author

Single molecule real-time DNA sequencing of HLA genes at ultra-high resolution from 126 International HLA and Immunogenetics Workshop cell lines.

HLA·2017
Same author

Recipient/donor HLA and CMV matching in recipients of T-cell-depleted unrelated donor haematopoietic cell transplants.

Bone marrow transplantation·2017
Same author

Variations in practice in UK transplant centers: results of a related donor care survey.

Bone marrow transplantation·2016
Same author

[Treatment of umbilical cord blood units with interleukin-15 improves hematopoietic stem cell engraftment].

Cirugia y cirujanos·2016
Same author

Further diversity of the 5' promoter region of the MHC class I-related chain B gene.

International journal of immunogenetics·2015

Reference Strand Mediated Conformation Analysis (RSCA) provides high-resolution HLA typing by analyzing DNA conformation. This novel method accurately identifies HLA alleles, even those differing by a single nucleotide.

Area of Science:

  • Immunogenetics
  • Molecular Biology
  • Biotechnology

Background:

  • Human Leukocyte Antigen (HLA) typing is crucial for transplantation and disease association studies.
  • Conventional sequence-based methods can be time-consuming and may struggle with high-resolution typing.
  • A need exists for high-throughput, accurate HLA typing techniques.

Purpose of the Study:

  • To introduce and evaluate Reference Strand Mediated Conformation Analysis (RSCA) as a novel HLA typing method.
  • To demonstrate the high resolution and throughput capabilities of RSCA for HLA class I and II genes.
  • To validate RSCA's reproducibility and ability to identify new alleles and resolve complex heterozygous combinations.

Main Methods:

  • RSCA utilizes locus-specific fluorescent-labeled DNA to alter the conformation of target DNA.

Related Experiment Videos

  • Conformation-dependent DNA mobility is measured using polyacrylamide gel electrophoresis (AGE).
  • Laser-based instrumentation, computer software, and internal DNA markers ensure accurate discrimination of alleles differing by a single nucleotide.

    • Main Results:

    • RSCA successfully typed HLA class I and II genes with high resolution.
    • The method demonstrated high sample throughput compared to conventional techniques.
    • Blind studies confirmed RSCA's reproducibility, ability to identify novel alleles, and resolve ambiguous heterozygous calls.

    Conclusions:

    • RSCA is a powerful and accurate conformational method for high-resolution HLA typing.
    • The technique offers significant advantages in throughput and discriminatory power.
    • RSCA represents a valuable advancement for immunogenetic research and clinical applications.