Histocompatibility screening by molecular techniques

Use of polymerase chain reaction products and heteroduplex formation

R. J. Rubocki, James Lowell Wisecarver, D. D. Hook, S. M. Cox, K. W. Beisel

Research output: Contribution to journalArticle

6 Citations (Scopus)

Abstract

Novel molecular approaches have recently become available allowing improved major histocompatibility complex (MHC) matching of potential allogeneic bone marrow donors and recipients. Current cellular and serological assays are hindered by aberrant cell populations and limited reagents which only detect an individuals' phenotype. Therefore, a molecular screening protocol which discriminates at the genotypic level would be advantageous. Here we describe a two-step DNA-based approach that can be applied to large-scale screening of potential donors. A primary screen, utilizing polymerase chain reaction (PCR), reduces the potential donor population, whereas a secondary or fine resolution screen uses DNA heteroduplex analysis to determine identity or non-identity at specific loci. Heteroduplex analysis generates a DNA migration pattern that is unique for alleles at a given locus, and is more sensitive than serology in discriminating among individuals. Here we demonstrate the potential feasibility of this approach by analyzing results at one MHC locus, HLA-DQ. Since this method does not rely on typing sera or viable lymphocytes, it is not subject to the variability found in the traditional methods. In contrast to traditional methods, these molecular techniques can provide the critical information needed to select a potential bone marrow donor.

Original languageEnglish (US)
Pages (from-to)337-341
Number of pages5
JournalJournal of Clinical Laboratory Analysis
Volume6
Issue number5
StatePublished - Jan 1 1992

Fingerprint

Histocompatibility
Polymerase chain reaction
Reaction products
Heteroduplex Analysis
Screening
Bone
Nucleic Acid Heteroduplexes
HLA-DQ Antigens
Major Histocompatibility Complex
Polymerase Chain Reaction
Lymphocytes
DNA
Bone Marrow
Donor Selection
Assays
Cells
Serology
Population
Alleles
Phenotype

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)

Cite this

Histocompatibility screening by molecular techniques : Use of polymerase chain reaction products and heteroduplex formation. / Rubocki, R. J.; Wisecarver, James Lowell; Hook, D. D.; Cox, S. M.; Beisel, K. W.

In: Journal of Clinical Laboratory Analysis, Vol. 6, No. 5, 01.01.1992, p. 337-341.

Research output: Contribution to journalArticle

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