A matrix-assisted laser desorption/ionization time-of-flight based method for screening the 1691G --> A mutation in the factor V gene

Blood Coagulation & Fibrinolysis : an International Journal in Haemostasis and Thrombosis
K HungB Guo

Abstract

A point mutation, 1691 G --> A in the coagulation factor V gene results in an Arg506 --> Gln amino acid substitution in the factor V molecule. This mutation, defined as factor VLEIDEN, results in activated protein C (APC) resistance and is the most common genetic risk factor for familial thrombophilia. A new mini-sequencing method using matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry was developed for the screening of the 1691G --> A substitution in factor V. In this method, a fragment of genomic DNA containing the 1691st base is first amplified, followed by mini-sequencing in the presence of dGTP and ddATP, ddCTP, and ddTTP. In this manner, the primer is extended by one base from one allele and two bases from the other allele. The extended products are analyzed using MALDI-TOF mass spectrometry. The base at position 1691 is identified based on the number of nucleotides added. We have used this method to genotype 16 APC-resistant patients previously identified by conventional methods and 11 normal control samples. The genotypes of all samples were correctly identified. This method is accurate, fast, and potentially allows for simultaneous multiplex genotyping of a number of mutation site...Continue Reading

References

Feb 24, 1995·The Journal of Biological Chemistry·M KalafatisK G Mann
Apr 1, 1997·Arteriosclerosis, Thrombosis, and Vascular Biology·M Kalafatis, K G Mann
Dec 16, 1998·Nature Biotechnology·P RossL Haff
Feb 12, 1999·Arteriosclerosis, Thrombosis, and Vascular Biology·M KalafatisK G Mann
May 26, 1999·Proceedings of the National Academy of Sciences of the United States of America·T J GriffinL M Smith
Jun 28, 2000·Nucleic Acids Research·X SunB Guo

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Citations

Apr 1, 2003·Mass Spectrometry Reviews·Jörg Tost, Ivo G Gut
May 10, 2012·Journal of Proteomics·Dominique RichterEduard F Stange
Feb 18, 2010·American Journal of Hematology·Ashkan EmadiJodi B Segal
Sep 26, 2003·Clinical and Applied Thrombosis/hemostasis : Official Journal of the International Academy of Clinical and Applied Thrombosis/Hemostasis·Ana-Maria SimundicMario Stefanovic
Feb 27, 2010·Diagnostic Molecular Pathology : the American Journal of Surgical Pathology, Part B·Shangbin YangHaifeng M Wu

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