Easy, accurate and reliable screening for SNPs by ion pair/reverse phase HPLC: simultaneous detection of factor V G1691A, prothrombin G20210A and methylenetetrahydrofolate reductase C677T variants.
Clin Lab
; 49(7-8): 313-8, 2003.
Article
en En
| MEDLINE
| ID: mdl-12908731
ABSTRACT
We have set up allele specific PCR in combination with high-performance liquid chromatography as a method to simultaneously detect three SNPs, factor V (G1691A), factor II/prothrombin (G20210A) and methylenetetrahydrofolate reductase (C677T) genes, all suspected to be inherited risk factors for deep vein thrombosis. Allele specific multiplex PCR technique was used to generate PCR products of different sizes which were specific for each genotype analyzed in this study. PCR products were analyzed on the WAVE Nucleic Acid Analysis platform by means of a gradient ion pair/reverse phase chromatography without any further purification. This method allows concurrent genotyping of the PCR products of the three mutations within 8 minutes, clearly reduces working time and costs in a routine laboratory, and can easily be adapted to the analysis of any other SNP or the combination of different other SNPs. The assay established here has been validated on more than a hundred samples and gave correct and highly reproducible intra- and interday results, allowing the application of this easy, accurate and reliable test in routine settings and in clinical trials and epidemiological surveys.
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Base de datos:
MEDLINE
Asunto principal:
Factor V
/
Protrombina
/
Pruebas Genéticas
/
Cromatografía Líquida de Alta Presión
/
Polimorfismo de Nucleótido Simple
/
Metilenotetrahidrofolato Reductasa (NADPH2)
Tipo de estudio:
Diagnostic_studies
/
Prognostic_studies
/
Risk_factors_studies
/
Screening_studies
Idioma:
En
Revista:
Clin Lab
Asunto de la revista:
TECNICAS E PROCEDIMENTOS DE LABORATORIO
Año:
2003
Tipo del documento:
Article