Rapid high-resolution melting method to identify human leukocyte antigen-G (HLA-G) 3' untranslated region polymorphism +3142C/G (rs1063320).
Journal
Molecular genetics & genomic medicine
ISSN: 2324-9269
Titre abrégé: Mol Genet Genomic Med
Pays: United States
ID NLM: 101603758
Informations de publication
Date de publication:
11 2021
11 2021
Historique:
revised:
30
01
2020
received:
02
10
2019
accepted:
05
03
2020
pubmed:
5
10
2021
medline:
5
4
2022
entrez:
4
10
2021
Statut:
ppublish
Résumé
HLA-G is a non-classical class I gene of the human Major Histocompatibility encoding molecules with immune-modulatory properties. Expression of HLA-G is being largely studied in pathological conditions, such as tumors, viral infections, inflammation, and autoimmune diseases, grafted tissues, among others. HLA-G +3142C/G (rs1063320: dbSNP database) polymorphism is located in 3' UTR of HAL-G and plays a key role in determining the magnitude of gene and protein expression. The detection of HLA-G +3142C/G polymorphism in the most published report is done through polymerase chain reaction followed by enzymatic digestion. Therefore, it is so interesting to develop a rapid and sensitive assay to genotype HLA-G +3142C/G polymorphism. High-resolution melt analysis (HRM) is a technology that is based on the analysis of the melting profile of PCR products through gradual temperature increase. The aim of this work is to apply high-resolution melt method for genotyping the HLA-G +3142C/G polymorphism. DNA from 118 individuals was extracted from whole blood with QIAamp We were able to recognize the three genotypes with similar accuracy than DNA sequencing using high resolution melting method. Hardy-Weinberg equilibrium test shows that our population is in equilibrium for the studied SNP. Genotypes frequencies of +3142C/G polymorphism in Tunisian general population are 0.475 for heterozygote G/C, 0.186 for homozygote G/G and 0.339 for homozygote C/C. HRM is a cost-effective method suitable for SNP genotyping.
Sections du résumé
BACKGROUND
HLA-G is a non-classical class I gene of the human Major Histocompatibility encoding molecules with immune-modulatory properties. Expression of HLA-G is being largely studied in pathological conditions, such as tumors, viral infections, inflammation, and autoimmune diseases, grafted tissues, among others. HLA-G +3142C/G (rs1063320: dbSNP database) polymorphism is located in 3' UTR of HAL-G and plays a key role in determining the magnitude of gene and protein expression. The detection of HLA-G +3142C/G polymorphism in the most published report is done through polymerase chain reaction followed by enzymatic digestion. Therefore, it is so interesting to develop a rapid and sensitive assay to genotype HLA-G +3142C/G polymorphism. High-resolution melt analysis (HRM) is a technology that is based on the analysis of the melting profile of PCR products through gradual temperature increase. The aim of this work is to apply high-resolution melt method for genotyping the HLA-G +3142C/G polymorphism.
METHODS
DNA from 118 individuals was extracted from whole blood with QIAamp
RESULTS
We were able to recognize the three genotypes with similar accuracy than DNA sequencing using high resolution melting method. Hardy-Weinberg equilibrium test shows that our population is in equilibrium for the studied SNP. Genotypes frequencies of +3142C/G polymorphism in Tunisian general population are 0.475 for heterozygote G/C, 0.186 for homozygote G/G and 0.339 for homozygote C/C.
CONCLUSION
HRM is a cost-effective method suitable for SNP genotyping.
Identifiants
pubmed: 34605219
doi: 10.1002/mgg3.1817
pmc: PMC8606219
doi:
Substances chimiques
3' Untranslated Regions
0
HLA-G Antigens
0
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
e1817Informations de copyright
© 2021 The Authors. Molecular Genetics & Genomic Medicine published by Wiley Periodicals LLC.
Références
Gene. 2013 Jan 1;512(1):108-12
pubmed: 23063736
Taiwan J Obstet Gynecol. 2017 Jun;56(3):276-280
pubmed: 28600033
Tissue Antigens. 2011 Jun;77(6):540-5
pubmed: 21395561
Mol Genet Genomic Med. 2021 Nov;9(11):e1817
pubmed: 34605219
J Immunol Res. 2014;2014:359748
pubmed: 24800261
Int J Mol Sci. 2017 Nov 05;18(11):
pubmed: 29113092
Nat Rev Genet. 2008 May;9(5):356-69
pubmed: 18398418
Am J Hum Genet. 2007 Oct;81(4):829-34
pubmed: 17847008
Immunol Lett. 2016 Dec;180:24-30
pubmed: 27771469
Nat Commun. 2018 Jul 27;9(1):2941
pubmed: 30054458
Tissue Antigens. 2009 Oct;74(4):308-13
pubmed: 19775370
Clin Chem. 2003 Jun;49(6 Pt 1):853-60
pubmed: 12765979
Tissue Antigens. 2014 Apr;83(4):260-6
pubmed: 24580026
Pediatr Rheumatol Online J. 2017 Feb 1;15(1):10
pubmed: 28143491
Clin Chem. 2004 Jul;50(7):1156-64
pubmed: 15229148
J Clin Pathol. 2008 Apr;61(4):487-93
pubmed: 17908804
Semin Cancer Biol. 2007 Dec;17(6):451-8
pubmed: 17716911
Tissue Antigens. 2012 Sep;80(3):259-62
pubmed: 22775767
Hum Immunol. 2017 Feb;78(2):172-178
pubmed: 28040535
Nat Rev Genet. 2005 Feb;6(2):95-108
pubmed: 15716906
Tissue Antigens. 2013 Oct;82(4):286-7
pubmed: 24032992
PLoS Comput Biol. 2012;8(12):e1002822
pubmed: 23300413
Inflamm Allergy Drug Targets. 2012 Dec;11(6):448-63
pubmed: 22931388
Clin Chem. 2008 Dec;54(12):2055-8
pubmed: 19042988
Immunol Res. 2016 Aug;64(4):961-8
pubmed: 26754763