Identification of specific BRCA1 and BRCA2 variants by DHPLC

被引:0
作者
Gross, E [1 ]
Arnold, N [1 ]
Pfeifer, K [1 ]
Bandick, K [1 ]
Kiechle, M [1 ]
机构
[1] Univ Kiel, Dept Obstet & Gynaecol, D-24105 Kiel, Germany
关键词
DHPLC analysis; BRCA1; BRCA2; mutation detection; cancer;
D O I
10.1002/1098-1004(200010)16:4<345::AID-HUMU7>3.0.CO;2-#
中图分类号
Q3 [遗传学];
学科分类号
071007 ; 090102 ;
摘要
Denaturing high performance liquid chromatography (DHPLC) is generating increasing interest in clinical genetics as a reliable tool for the analysis of genetic alterations. In the work presented here our intentions were to optimize primer design and DHPLC analysis conditions for a qualitative detection of BRCA1 and BRCA2 variations. The BRCA1 and BRAC2 genes display a high proportion of polymorphisms. Sequencing efforts geared towards the distinction of tumor-related mutations and benign variants still remain time-consuming and expensive. DHPLC elution profiles, however, permit the correlation of a characteristic chromatographic profile with a specific sequence alteration. In this study we evaluate the sensitivity of DHPLC for the identification of unique polymorphisms, which are frequent in the Caucasian population, in lieu of sequence analysis. The complete BRCA1 gene and parts of BRCA2 were examined. In the case of BRCA1, 431 out of 432 heterozygotes were identified correctly. Tn addition, 18 new profiles were identified which had not been detected previously in our studies and which represented new mutations or rare polymorphisms. For BRCA2, 135 out of 137 simple sequence variants were classified correctly. In addition, six new profiles were identified which represented new mutations or rare polymorphisms. Hum Mutat 16:345-353, 2000. (C) 2000 Wiley-Liss, Inc.
引用
收藏
页码:345 / 353
页数:9
相关论文
共 16 条
  • [1] [Anonymous], 1998, Primer 3
  • [2] Arnold N, 1999, HUM MUTAT, V14, P333, DOI 10.1002/(SICI)1098-1004(199910)14:4<333::AID-HUMU9>3.3.CO
  • [3] 2-3
  • [4] Comparison of fluorescent single-strand conformation polymorphism analysis and denaturing high-performance liquid chromatography for detection of EXT1 and EXT2 mutations in hereditary multiple exostoses
    Dobson-Stone, C
    Cox, RD
    Lonie, L
    Southam, L
    Fraser, M
    Wise, C
    Bernier, F
    Hodgson, S
    Porter, DE
    Simpson, AHRW
    Monaco, AP
    [J]. EUROPEAN JOURNAL OF HUMAN GENETICS, 2000, 8 (01) : 24 - 32
  • [5] A comparison of BRCA1 mutation analysis by direct sequencing, SSCP and DHPLC
    Gross, E
    Arnold, N
    Goette, J
    Schwarz-Boeger, U
    Kiechle, M
    [J]. HUMAN GENETICS, 1999, 105 (1-2) : 72 - 78
  • [6] HAYWARDLESTER A, 1996, GENE QUANTIFICATION, P44
  • [7] Jones AC, 1999, CLIN CHEM, V45, P1133
  • [8] Denaturing high performance liquid chromatography (DHPLC) used in the detection of germline and somatic mutations
    Liu, WG
    Smith, DI
    Rechtzigel, KJ
    Thibodeau, SN
    James, CD
    [J]. NUCLEIC ACIDS RESEARCH, 1998, 26 (06) : 1396 - 1400
  • [9] Nickerson ML, 2000, HUM MUTAT, V16, P68, DOI 10.1002/1098-1004(200007)16:1<68::AID-HUMU12>3.0.CO
  • [10] 2-U