Alternative methods to a TaqMan assay to detect a tri-allelic single nucleotide polymorphism rs757210 in the HNF1ß gene.
Clin Chem Lab Med
; 50(2): 279-84, 2011 Oct 21.
Article
en En
| MEDLINE
| ID: mdl-22022982
ABSTRACT
BACKGROUND:
Several studies report difficulties in genotyping HNF1ß rs757210 using TaqMan probes. This is possibly due to the tri-allelic nature of this single nucleotide polymorphism (SNP). The aim of the present research was to develop alternative methods for genotyping rs757210.METHODS:
Pyrosequencing and high resolution melting analysis of small amplicons (HRM) were developed and tested in panels of type 2 diabetes mellitus patients (n=258) and healthy blood donors (n=183). Results were confirmed by Sanger sequencing.RESULTS:
With pyrosequencing, allele frequencies for the A, G and C allele of 0.42, 0.56, 0.02 and 0.37, 0.62, 0.01 were established in the panel of type 2 diabetes mellitus patients and healthy blood donors, respectively. Similar results were found using the more routinely available HRM method. Results for pyrosequencing and HRM were in 99.6% concordance.CONCLUSIONS:
Pyrosequencing and HRM can be used to genotype the tri-allelic SNP rs757210 in the HNF1ß gene and have the advantage over the commercially available TaqMan analysis that they can determine the rare C-allele variant.
Texto completo:
1
Bases de datos:
MEDLINE
Asunto principal:
Polimorfismo de Nucleótido Simple
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Alelos
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Factor Nuclear 1-beta del Hepatocito
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Técnicas de Genotipaje
Límite:
Female
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Humans
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Male
Idioma:
En
Revista:
Clin Chem Lab Med
Asunto de la revista:
QUIMICA CLINICA
/
TECNICAS E PROCEDIMENTOS DE LABORATORIO
Año:
2011
Tipo del documento:
Article
País de afiliación:
Países Bajos