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A new PCR-SSP method for HLA DR-DQ risk assessment for celiac disease.

Lavant, Ewa H; Agardh, Daniel LU ; Ramelius, Anita LU and Carlson, Joyce LU (2011) In Clinica Chimica Acta 412. p.782-784
Abstract
BACKGROUND: Susceptibility to celiac disease is essentially restricted to carriers of specific HLA DQA1 and DQB1 alleles. We have developed a semi-automated sequence specific primer (SSP) PCR method for clinical HLA typing and compared the test results with those from a commercial method. METHODS: Primers for each DQA1 and DQB1 allele group were included in our PCR-SSP reaction to allow differentiation of homozygous from heterozygous carriers of risk alleles. Primers detecting the tightly linked DRB1*04, *03, *07 and *09 alleles were included to resolve potentially ambiguous results. Fluorescently labeled PCR products of 119 clinical samples were analyzed by capillary electrophoresis, and results were compared to those previously obtained... (More)
BACKGROUND: Susceptibility to celiac disease is essentially restricted to carriers of specific HLA DQA1 and DQB1 alleles. We have developed a semi-automated sequence specific primer (SSP) PCR method for clinical HLA typing and compared the test results with those from a commercial method. METHODS: Primers for each DQA1 and DQB1 allele group were included in our PCR-SSP reaction to allow differentiation of homozygous from heterozygous carriers of risk alleles. Primers detecting the tightly linked DRB1*04, *03, *07 and *09 alleles were included to resolve potentially ambiguous results. Fluorescently labeled PCR products of 119 clinical samples were analyzed by capillary electrophoresis, and results were compared to those previously obtained from the DELFIA® Type 1 Diabetes Genetic Predisposition assay. RESULTS: The risk assessment derived from the two methods was 100% concordant. One previously unreported haplotype was detected and haplotype assignments in two of the 119 samples were improved from previous reports. CONCLUSIONS: The use of three PCR reactions and a single electrophoretic step for DQA1, DQB1 and DRB1 typing provides distinction of celiac disease associated alleles and their homo- or heterozygous status. This multiplex analysis reduces reagent costs, personnel and instrument time, while enabling improved allelic assignment through HLA-DR-DQ haplotype association. (Less)
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author
organization
publishing date
type
Contribution to journal
publication status
published
subject
in
Clinica Chimica Acta
volume
412
pages
782 - 784
publisher
Elsevier
external identifiers
  • wos:000288634500018
  • pmid:21219892
  • scopus:79951724617
ISSN
0009-8981
DOI
10.1016/j.cca.2010.12.033
language
English
LU publication?
yes
id
3798564a-47ad-412e-96bd-0f9932adb7e5 (old id 1777649)
alternative location
http://www.ncbi.nlm.nih.gov/pubmed/21219892?dopt=Abstract
date added to LUP
2011-02-01 12:15:07
date last changed
2017-10-22 04:51:59
@article{3798564a-47ad-412e-96bd-0f9932adb7e5,
  abstract     = {BACKGROUND: Susceptibility to celiac disease is essentially restricted to carriers of specific HLA DQA1 and DQB1 alleles. We have developed a semi-automated sequence specific primer (SSP) PCR method for clinical HLA typing and compared the test results with those from a commercial method. METHODS: Primers for each DQA1 and DQB1 allele group were included in our PCR-SSP reaction to allow differentiation of homozygous from heterozygous carriers of risk alleles. Primers detecting the tightly linked DRB1*04, *03, *07 and *09 alleles were included to resolve potentially ambiguous results. Fluorescently labeled PCR products of 119 clinical samples were analyzed by capillary electrophoresis, and results were compared to those previously obtained from the DELFIA® Type 1 Diabetes Genetic Predisposition assay. RESULTS: The risk assessment derived from the two methods was 100% concordant. One previously unreported haplotype was detected and haplotype assignments in two of the 119 samples were improved from previous reports. CONCLUSIONS: The use of three PCR reactions and a single electrophoretic step for DQA1, DQB1 and DRB1 typing provides distinction of celiac disease associated alleles and their homo- or heterozygous status. This multiplex analysis reduces reagent costs, personnel and instrument time, while enabling improved allelic assignment through HLA-DR-DQ haplotype association.},
  author       = {Lavant, Ewa H and Agardh, Daniel and Ramelius, Anita and Carlson, Joyce},
  issn         = {0009-8981},
  language     = {eng},
  pages        = {782--784},
  publisher    = {Elsevier},
  series       = {Clinica Chimica Acta},
  title        = {A new PCR-SSP method for HLA DR-DQ risk assessment for celiac disease.},
  url          = {http://dx.doi.org/10.1016/j.cca.2010.12.033},
  volume       = {412},
  year         = {2011},
}