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Human leukocyte antigen typing using buccal swabs as accurate and non-invasive substitute for venipuncture in children at risk for celiac disease.

Author(s): Adriaanse MP, Vreugdenhil AC, Vastmans V, Groeneveld L, Molenbroeck S, Schott DA, Voorter CE, Tilanus MG

Publication: J Gastroenterol Hepatol, 2016, Vol. 31, Page 1711-1716

PubMed ID: 26945703 PubMed Review Paper? No

Purpose of Paper

Conclusion of Paper

Studies

1. Study Purpose

   This study investigated the ability to collect sufficient DNA from buccal cell swab specimens to perform celiac disease-related HLA typing and compared results to whole blood collected by venipuncture. Specimens were collected from 77 patients (age range 2.3–22.4 years, mean 13.6 years, 58.4% women) at high-risk for celiac disease between August 2012 and December 2013. Collection was performed at the outpatient clinic for (n=55, 71.4%) patients or at home by the patient or parents (n=22, 28.6%). Buccal swab collection was performed at least 30 minutes after a meal by swabbing the inside of the cheek with a dry cotton swab for 15 seconds in each corner of the mouth (n=4). Collections performed at home were mailed and received within 1–3 days. Buccal swabs were stored at room temperature until DNA extraction within 4 weeks. Two swabs per participant were used for DNA elution on a QIamp column. Whole blood was collected by venipuncture in EDTA tubes from all 77 patients at the clinic and DNA was isolated using the QIAmp procedure. Typing of HLA-DRB1, HLA-DQA1, and HLADQB1 was performed by a sequence-specific oligonucleotide hybridization method and HLA-DQ8 identification was performed by sequence-based typing. To study whether buccal swabs provide sufficient DNA for HLA typing in infants, buccal cell specimens were collected from six healthy children under one year of age.

   Summary of Findings:

   Median DNA yield from buccal cell swabs was 4.68 (range 2.34–8.10) μg per two buccal swabs with a median OD260/OD280 ration of 2.09 (2.01–2.16). All specimens yielded a DNA of sufficient quantity and purity to perform HLA typing, but the purity of buccal cell DNA was significantly lower compared to DNA from blood specimens (2.09 versus 1.92, P<0.001). The HLA class-II typing results for buccal cell-derived DNA and blood-derived DNA were identical for all participants. DNA yields were comparable in specimens collected at home to those collected at the outpatient clinic and HLA typing results on buccal swabs corresponded with the results from the blood specimens. Specimens from infants yielded a sufficient amount and purity of DNA for HLA typing (median DNA yield of 5.34 μg per two swabs and median OD260/OD280 ratio of 2.13).

   Biospecimens

   • Bodily Fluid - Blood
   • Cell - Buccal

   Preservative Types

   • Other Preservative

   Diagnoses:

   • Other diagnoses
   • Normal

   Platform:

   Analyte	Technology Platform
   DNA	DNA sequencing

   Pre-analytical Factors:

   Classification	Pre-analytical Factor	Value(s)
   Biospecimen Acquisition	Method of fluid acquisition	Cheek rubbing during collection Venipuncture
   Biospecimen Acquisition	Biospecimen location	Buccal cell Venous blood
   Preaquisition	Patient age	<1 year 2.3–22.4 years
   Biospecimen Acquisition	Locale of biospecimen collection	Collected at home Collected in clinic

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