TY - JOUR
T1 - Enzymatic mutation detection (EMD) of novel mutations (R565X and R1523X) in the FBN1 gene of patients with Marfan syndrome using T4 endonuclease VII
AU - Youil, Rima
AU - Toner, Timothy J.
AU - Bull, Evelyn
AU - Bailey, Anne L.
AU - Earl, Christopher D.
AU - Dietz, Harry C.
AU - Montgomery, Robert Avery
PY - 2000/7
Y1 - 2000/7
N2 - 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. Copyright Wiley-Liss, Inc.
AB - 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. Copyright Wiley-Liss, Inc.
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U2 - 10.1002/1098-1004(200007)16:1<92::AID-HUMU24>3.0.CO;2-1
DO - 10.1002/1098-1004(200007)16:1<92::AID-HUMU24>3.0.CO;2-1
M3 - Article
C2 - 10874320
AN - SCOPUS:0034223108
SN - 1059-7794
VL - 16
SP - 92
EP - 93
JO - Human mutation
JF - Human mutation
IS - 1
ER -