Evaluation of a High Throughput Method for the Detection of Mutations Associated with Thrombosis and Hereditary Hemochromatosis in Brazilian Blood Donors

Autor: Juliana Vieira dos Santos Bianchi, Ester Cerdeira Sabino, Vivian Dionisio Tavares Niewiadonski, Cesar de Almeida-Neto, Nelson Gaburo
Rok vydání: 2015
Předmět:
Male
Pathology
Gene Expression
lcsh:Medicine
Blood Donors
Gastroenterology
Gene Frequency
lcsh:Science
Oligonucleotide Array Sequence Analysis
Venous Thrombosis
Multidisciplinary
medicine.diagnostic_test
Middle Aged
Thrombosis
Venous thrombosis
Hereditary hemochromatosis
Female
Prothrombin
Hemochromatosis
Brazil
Research Article
Adult
congenital
hereditary
and neonatal diseases and abnormalities
medicine.medical_specialty
Hyperhomocysteinemia
Genotype
Real-Time Polymerase Chain Reaction
Polymorphism
Single Nucleotide
Sensitivity and Specificity
Internal medicine
parasitic diseases
medicine
Humans
Genetic Testing
Hemochromatosis Protein
Genotyping
Alleles
Methylenetetrahydrofolate Reductase (NADPH2)
Genetic testing
business.industry
Histocompatibility Antigens Class I
lcsh:R
Factor V
Membrane Proteins
nutritional and metabolic diseases
medicine.disease
High-Throughput Screening Assays
Genotype frequency
lcsh:Q
business
Zdroj: PLoS ONE, Vol 10, Iss 5, p e0125460 (2015)
PLoS ONE
ISSN: 1932-6203
DOI: 10.1371/journal.pone.0125460
Popis: Background The aim of this study was to evaluate the OpenArray platform for genetic testing of blood donors and to assess the genotype frequencies of nucleotide-polymorphisms (SNPs) associated with venous thrombosis (G1691A and G20210A), hyperhomocysteinemia (C677T, A1298C), and hereditary hemochromatosis (C282Y, H63D and S65C) in blood donors from Sao Paulo, Brazil. Methods We examined 400 blood donor samples collected from October to November 2011. The SNPs were detected using OpenArray technology. The blood samples were also examined using a real-time PCR–FRET system to compare the results and determine the accuracy of the OpenArray method. Results We observed 100% agreement in all assays tested, except HFE C282Y, which showed 99.75% agreement. The HFE C282Y assay was further confirmed through direct sequencing, and the results showed that OpenArray analysis was accurate. The calculated frequencies of each SNP were FV G1691A 98.8% (G/G), 1.2% (G/A); FII G2021A 99.5% (G/G), 0.5% (G/A); MTHFR C677T 45.5% (C/C), 44.8% (C/T), 9.8% (T/T); MTHFR A1298C 60.3% (A/A), 33.6% (A/C), 6.1% (C/C); HFE C282Y 96%(G/G), 4%(G/A), HFE H63D 78.1%(C/C), 20.3% (C/G), 1.6% (G/G); and HFE S65C 98.1% (A/A), 1.9% (A/T). Conclusion Taken together, these results describe the frequencies of SNPs associated with diseases and are important to enhance our current knowledge of the genetic profiles of Brazilian blood donors, although a larger study is needed for a more accurate determination of the frequency of the alleles. Furthermore, the OpenArray platform showed a high concordance rate with standard FRET RT-PCR.
Databáze: OpenAIRE
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