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Identification Of Fviii Gene Mutations In Patients Withhemophilia A Using New Combinatorial Sequencing By Hybridization | 6082
ISSN: 2155-9872

Journal of Analytical & Bioanalytical Techniques
Open Access

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Identification of Fviii Gene Mutations in Patients Withhemophilia a Using New Combinatorial Sequencing by Hybridization

International Conference & Exihibition On Analytical and Bioanalytical Techniques - 2010

Massimiliano Chetta

ScientificTracks Abstracts: J Anal Bioanal Techniques

DOI: 10.4172/2155-9872.1000001

Abstract
Standard methods of mutation detection are time consuming in Hemophilia A (HA) rendering their application unavailable in some analysis such as prenatal diagnosis. To evaluate the feasibility of combinatorial sequencing-by-hybridization (cSBH) as an alternative and reliable tool for mutation detection in FVIII gene. We have applied a new method of cSBH that uses two different colors for detection of multiple point mutations in the FVIII gene. The 26 exons encompassing the HA gene were analyzed in 7 newly diagnosed Italian patients and in 19 previously characterized individuals with FVIII de� ciency. Data show that, when solution- phase TAMRA and QUASAR labeled 5-mer oligonucleotide sets mixed with unlabeled target PCR templates are co-hybridized in the presence of DNA ligase to universal 6-mer oligonucleotide probe-based arrays, a number of mutations can be successfully detected. The technique was reliable also in identifying a mutant FVIII allele in an obligate heterozygote. A novel missense mutation (Leu1843Thr) in exon 16 and three novel neutral polymorphisms are presented with an updated protocol for 2-color cSBH.cSBH is a reliable tool for mutation detection in FVIII gene and may represent a complementary method for the genetic screening of HA patients.
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