Screening for exonic copy number mutations at MSH2 and MLH1 by MAPH

被引：0
作者：
Seyed Mohammad Akrami
Malcolm G. Dunlop
Susan M. Farrington
Ian M. Frayling
Fiona MacDonald
John F. Harvey
John A. L. Armour
机构：
[1] University of Nottingham,Institute of Genetics
[2] Queen’s Medical Centre,Colon Cancer Genetics Group, Division of Oncology
[3] University of Edinburgh,Molecular Genetics Laboratory, Department of Medical Genetics
[4] Western General Hospital,West Midland Regional Genetics Laboratory
[5] Addenbrooke’s Hospital,Wessex Regional Genetics Laboratory
[6] Birmingham Women’s Hospital NHS Trust,Tehran University of Medical Sciences
[7] Salisbury District Hospital,Institute of Medical Genetics
[8] EMRC,undefined
[9] University Hospital of Wales,undefined
来源：
Familial Cancer | 2005年 / 4卷
关键词：
colorectal cancer; copy number; HNPCC; MAPH; MLPA;
D O I：
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学科分类号：
摘要：
Background: Exonic deletions in MSH2 and MLH1 are significant contributors to the mutation spectrum in HNPCC, and heterozygous changes in exon copy number are not detected by conventional mutation screening methods. Aims: We aimed to develop methods for screening copy number changes in all the exons of the MLH1 and MSH2 genes using a single multiplex amplifiable probe hybridisation (MAPH) assay. Methods: We developed a probe set consisting of probes from the 19 exons of MLH1 and 16 exons of MSH2, and 3 control probes, and applied it to screening for deletions and duplications using fluorescent detection of amplified fragments. Results: We tested 73 DNA samples from controls and 50 from HNPCC patients in whom no point mutations had been found, and detected 10 copy number changes among the patient samples. A deletion of about 1.4 kb including exon 3 of MSH2 was confirmed by amplification of a junction fragment, and was shown to be the result of an unequal recombination between intronic Alu elements. Conclusions: MAPH can detect exonic copy number changes in MLH1 and MSH2 in DNA from HNPCC patients. Since finding an exonic deletion or duplication makes full sequence analysis unnecessary, it may be most cost-effective to pre-screen samples by MAPH or MLPA before screening for point mutations.
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页码：145 / 149
页数：4
相关论文
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