title: Enzymatic mutation detection (EMD) of novel mutations (R565X and R1523X) in the FBN1 gene of patients with Marfan syndrome using T4 endonuclease VII.
abstract: The Enzymatic Mutation Detection (EMDtrade mark) method is a streamlined and improved version of the original Enzymatic Cleavage of Mismatch (EMC) method. EMD is a fully homogeneous, rapid four step procedure that allows for detection and localization of mismatched or unmatched nucleotides within heteroduplex DNA. To test the utility of EMD for use in the screening of large and complex genes, the fibrillin 1 (FBN1) gene was scanned in a cohort of six patients diagnosed with connective tissue disorders. Four of the six patients were diagnosed with classic Marfan syndrome (MFS). The results were compared with a previous MDEtrade mark scanning of the same patient cohort. Two causative mutations, R565X and R1523X, were detected by EMD that were not detected by MDE. In both cases, the mutation resulted in premature termination of translation. In addition, several polymorphisms were detected by the enzymatic approach that failed detection by heteroduplex analysis. We propose that the EMD method is a sensitive and rapid approach to mutation detection in large genes such as FBN1.
authors.affiliation: Merck Research Laboratories, West Point, PA, 19486, USA. rima_youil@merck.com
authors.forename: R
authors.initials: R
authors.lastname: Youil
authors.forename: T J
authors.initials: TJ
authors.lastname: Toner
authors.forename: E
authors.initials: E
authors.lastname: Bull
authors.forename: A L
authors.initials: AL
authors.lastname: Bailey
authors.forename: C D
authors.initials: CD
authors.lastname: Earl
authors.forename: H C
authors.initials: HC
authors.lastname: Dietz
authors.forename: R A
authors.initials: RA
authors.lastname: Montgomery
mesh_terms: D000328:Adult; D019943:Amino Acid Substitution / Q000235:genetics*; D001120:Arginine / Q000235:genetics; D004706:Endodeoxyribonucleases; D005260:Female; D000071838:Fibrillin-1; D000071837:Fibrillins; D006801:Humans; D008382:Marfan Syndrome / Q000235:genetics*; D008840:Microfilament Proteins / Q000235:genetics* / Q000378:metabolism; D008875:Middle Aged; D009154:Mutation / Q000235:genetics*; D010865:Pilot Projects; D011933:Reagent Kits, Diagnostic; D016037:Single-Blind Method; D054392:Adipokines
chemical_list: D001120:Arginine; D004706:Endodeoxyribonucleases; D000071838:Fibrillin-1; D000071837:Fibrillins; D008840:Microfilament Proteins; D011933:Reagent Kits, Diagnostic; C000605722:FBN1 protein, human; D054392:Adipokines; C035571:endodeoxyribonuclease VII