Rapid, loop-mediated isothermal amplification detection of coeliac disease risk alleles
- Author(s)
- Erlichster, M; Tye-Din, JA; Varney, MD; Skafidas, E; Kwan, P;
- Details
- Publication Year 2018,Volume 20,Issue #3,Page 307-315
- Journal Title
- Journal of Molecular Diagnostics
- Publication Type
- Journal Article
- Abstract
- Human leukocyte antigen (HLA) genotyping has become a useful investigation in the diagnostic work-up of coeliac disease (CD), with utility in risk stratification and screening. However, broad application of this technology has been hindered by the cost and time burden of conventional laboratory-based assays. We have developed and validated CD-loop-mediated isothermal amplification (CD-LAMP), a LAMP assay, which enables rapid identification of the signature CD risk genotypes, HLA-DQ2.5, HLA-DQ8, HLA-DQ2.2, and HLA-DQA1*05. Sample-to-answer is achieved in approximately 65 minutes without DNA purification, thermal cycling, or specialized analytical equipment. CD-LAMP genotyping of samples was 100% concordant with accredited pathology genotyping on a panel of 40 blood and 20 saliva samples. In a panel of 100 purified DNA samples, genotyping of the high risk DQ2.5 genotype was 100% concordant with accredited pathology genotyping, with slightly reduced sensitivity for the DQ8 genotype (97.1%), and reduced specificity for the DQ8 (93.9%) and DQ2.2 genotypes (95.1%). CD-LAMP results are easily visualized, instrument free, through the addition of a DNA intercalating dye following amplification. Combined with point-of-care antibody testing, CD-LAMP may enable immediate, confident CD diagnosis at a low cost in the clinical setting.
- Publisher
- Elsevier
- Research Division(s)
- Immunology
- PubMed ID
- 29458095
- Publisher's Version
- https://doi.org/10.1016/j.jmoldx.2018.01.005
- Terms of Use/Rights Notice
- Refer to copyright notice on published article.
Creation Date: 2018-02-28 10:12:08
Last Modified: 2019-06-20 02:04:42