Sequence diversity within the HA-1 gene as detected by melting temperature assay without oligonucleotide probes

Affordable Access

Sequence diversity within the HA-1 gene as detected by melting temperature assay without oligonucleotide probes

Publisher
BioMed Central
Publication Date
Oct 04, 2005
Source
PMC
Keywords
Disciplines
  • Biology
  • Medicine
License
Unknown

Abstract

1471-2350-6-36.fm ral ss BioMed CentBMC Medical Genetics Open AcceTechnical advance Sequence diversity within the HA-1 gene as detected by melting temperature assay without oligonucleotide probes Claudio Graziano1,2, Massimo Giorgi1, Cecilia Malentacchi1, Pier Luigi Mattiuz1 and Berardino Porfirio*1 Address: 1Human Genetics Unit, Department of Clinical Physiopathology, University of Florence, Viale Gaetano Pieraccini 6, 50139 Firenze, Italy and 2Current address: U.O. e Cattedra di Genetica Medica, Policlinico S. Orsola-Malpighi, Via Massarenti 9, 40138 Bologna, Italy Email: Claudio Graziano - [email protected]; Massimo Giorgi - [email protected]; Cecilia Malentacchi - [email protected]; Pier Luigi Mattiuz - [email protected]; Berardino Porfirio* - [email protected] * Corresponding author Abstract Background: The minor histocompatibility antigens (mHags) are self-peptides derived from common cellular proteins and presented by MHC class I and II molecules. Disparities in mHags are a potential risk for the development of graft-versus-host disease (GvHD) in the recipients of bone marrow from HLA-identical donors. Two alleles have been identified in the mHag HA-1. The correlation between mismatches of the mHag HA-1 and GvHD has been suggested and methods to facilitate large-scale testing were afterwards developed. Methods: We used sequence specific primer (SSP) PCR and direct sequencing to detect HA-1 gene polymorphisms in a sample of 131 unrelated Italian subjects. We then set up a novel melting temperature (Tm) assay that may help identification of HA-1 alleles without oligonucleotide probes. Results: We report the frequencies of HA-1 alleles in the Italian population and the presence of an intronic 5 base-pair deletion associated with the immunogeneic allele HA-1H. We also detected novel variable sites with respect to the consensus sequence of HA-1 locus. Even though recombination/gene conversion events are documented, there is considerable linkage dis

Report this publication

Statistics

Seen <100 times