PMID- 23314539
OWN - NLM
STAT- MEDLINE
DCOM- 20140507
LR  - 20161018
IS  - 1437-4331 (Electronic)
IS  - 1434-6621 (Linking)
VI  - 51
IP  - 6
DP  - 2013 Jun
TI  - A multiplex assay to rapidly exclude HLA-DQ2.5 and HLA-DQ8 expression in patients 
      at risk for celiac disease.
PG  - 1191-8
LID - 10.1515/cclm-2012-0774 [doi]
AB  - BACKGROUND: Celiac disease (CD) is an inflammatory disorder of the small 
      intestine induced by gluten ingestion. CD has a strong genetic association with 
      human leukocyte antigen (HLA)-DQ2.5 and HLA-DQ8. The absence of HLA-DQ2.5 and 
      HLA-DQ8 has a strong negative predictive value for CD. Genetic screening of 
      HLA-DQ2.5 and HLA-DQ8 in patients at risk is of great value. METHODS: We 
      designed, developed, and validated a multiplex assay based on multiplex 
      ligation-dependent probe amplification (MLPA) technology, allowing the 
      simultaneous detection of DQA1*05-DQB1*02, encoding HLA-DQ2.5, and 
      DQA1*03-DQB1*03:02, encoding HLA-DQ8. The amplified products were separated and 
      identified using capillary electrophoresis. RESULTS: When compared with a 
      polymerase chain reaction followed by single-strand conformation polymorphism/ 
      heteroduplex analysis, one discrepancy was found. Sequencing analysis showed that 
      the developed MLPA assay result was correct. Furthermore, we demonstrated that 
      the MLPA method is able to distinguish between the heterozygote and homozygote 
      expression of HLA-DQ2.5 or HLA-DQ8. CONCLUSIONS: This study shows that it is 
      possible to rapidly and accurately screen for the absence of HLA-DQ2.5 and 
      HLA-DQ8 using MLPA, excluding patients at risk for CD for further serological or 
      histological follow-up. In addition, MLPA might be an accurate tool to screen for 
      other specific HLA types in the context of disease association in a diagnostic 
      laboratory setting.
FAU - van Beek, Ellen M
AU  - van Beek EM
AD  - Department of Clinical Chemistry, Hematology and Immunology, Diakonessenhuis 
      Utrecht, Utrecht, The Netherlands. evbeek@diakhuis.nl
FAU - Roelandse-Koop, Elianne A
AU  - Roelandse-Koop EA
FAU - Vijzelaar, Raymon
AU  - Vijzelaar R
FAU - Yilmaz, Rizkat
AU  - Yilmaz R
FAU - van Hoogstraten, Ingrid M W
AU  - van Hoogstraten IM
FAU - Schreurs, Marco W J
AU  - Schreurs MW
FAU - Verheul, Alice A M
AU  - Verheul AA
FAU - van Houte, Arend Jan
AU  - van Houte AJ
FAU - Kortlandt, Wouter
AU  - Kortlandt W
LA  - eng
PT  - Journal Article
PL  - Germany
TA  - Clin Chem Lab Med
JT  - Clinical chemistry and laboratory medicine
JID - 9806306
RN  - 0 (HLA-DQ Antigens)
RN  - 0 (HLA-DQ2 antigen)
RN  - 0 (HLA-DQ8 antigen)
SB  - IM
MH  - Celiac Disease/*diagnosis/genetics/*immunology
MH  - Electrophoresis, Capillary/methods
MH  - Genetic Predisposition to Disease
MH  - HLA-DQ Antigens/*analysis/*biosynthesis/genetics
MH  - Humans
MH  - Multiplex Polymerase Chain Reaction/*methods
MH  - Polymerase Chain Reaction/methods
MH  - Polymorphism, Single-Stranded Conformational
MH  - Risk Factors
MH  - Sequence Analysis, DNA
EDAT- 2013/01/15 06:00
MHDA- 2014/05/08 06:00
CRDT- 2013/01/15 06:00
PHST- 2012/09/27 00:00 [received]
PHST- 2012/11/17 00:00 [accepted]
PHST- 2013/01/15 06:00 [entrez]
PHST- 2013/01/15 06:00 [pubmed]
PHST- 2014/05/08 06:00 [medline]
AID - /j/cclm.ahead-of-print/cclm-2012-0774/cclm-2012-0774.xml [pii]
AID - 10.1515/cclm-2012-0774 [doi]
PST - ppublish
SO  - Clin Chem Lab Med. 2013 Jun;51(6):1191-8. doi: 10.1515/cclm-2012-0774.