J Med Genet 2001;38:824-833 ( December )

Interpreting epidemiological research: blinded comparison of methods used to estimate the prevalence of inherited mutations in BRCA1

Charis Eng^a, Lawrence C Brody^b, Teresa M U Wagner^c, Peter Devilee^d, Jan Vijg^e, Csilla Szabo^f, Sean V Tavtigian^g, Katharine L Nathanson^h, Elaine Ostranderⁱ, Thomas S Frank^g, on behalf of the Steering Committee of the Breast Cancer Information Core (BIC) Consortium*

^a Clinical Cancer Genetics and Human Cancer Genetics Programs, Comprehensive Cancer Center, and Division of Human Genetics, Department of Internal Medicine, The Ohio State University, Columbus, OH, USA and CRC Human Cancer Genetics Research Group, University of Cambridge, Cambridge, UK, ^b National Institutes of Health, Bethesda, MD, USA, ^c Division of Senology, Department of Obstetrics and Gynecology, University of Vienna, Austria^¶ , ^d Departments of Human Genetics and Pathology, Leiden University Medical Centre, Leiden, The Netherlands , ^e Cancer Therapy and Research Center and University of Texas Health Science Center, San Antonio, TX, USA^§ , ^f International Agency for Research on Cancer, Lyon, France, ^g Myriad Genetic Laboratories, Salt Lake City, UT, USA^** , ^h Department of Medicine, University of Pennsylvania, Philadelphia, PA, USA, ⁱ Clinical Research Division, Fred Hutchinson Cancer Research Center, Seattle, WA, USA 

Correspondence to: Professor Eng, Ohio State University Human Cancer Genetics Program, 420 W 12th Avenue, Suite 690 Tzagournis MRF, Columbus, OH 43210, USA, eng-1{at}medctr.osu.edu or ceng{at}hgmp.mrc.ac.uk

Revised version received 1 October 2001; Accepted for publication 2 October 2001

While sequence analysis is considered by many to be the most sensitive method of detecting unknown mutations in large genes such as BRCA1, most published estimates of the prevalence of mutations in this gene have been derived from studies that have used other methods of gene analysis. In order to determine the relative sensitivity of techniques that are widely used in research on BRCA1, a set of blinded samples containing 58 distinct mutations were analysed by four separate laboratories. Each used one of the following methods: single strand conformational polymorphism analysis (SSCP), conformation sensitive gel electrophoresis (CSGE), two dimensional gene scanning (TDGS), and denaturing high performance liquid chromatography (DHPLC). Only the laboratory using DHPLC correctly identified each of the mutations. The laboratory using TDGS correctly identified 91% of the mutations but produced three apparent false positive results. The laboratories using SSCP and CSGE detected abnormal migration for 72% and 76% of the mutations, respectively, but subsequently confirmed and reported only 65% and 60% of mutations, respectively. False negatives therefore resulted not only from failure of the techniques to distinguish wild type from mutant, but also from failure to confirm the mutation by sequence analysis as well as from human errors leading to misreporting of results. These findings characterise sources of error in commonly used methods of mutation detection that should be addressed by laboratories using these methods. Based upon sources of error identified in this comparison, it is likely that mutations in BRCA1 and BRCA2 are more prevalent than some studies have previously reported. The findings of this comparison provide a basis for interpreting studies of mutations in susceptibility genes across many inherited cancer syndromes.


Keywords: BRCA1; mutation detection; cancer genetics

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© 2001 by J Med Genet
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