Mattocks C, Tarpey P, Bobrow M, Whittaker J
Molecular Genetics Laboratory, Addenbrooke's Hospital, Cambridge, UK.
Hum Mutat. 2000 Nov;16(5):437-43. doi: 10.1002/1098-1004(200011)16:5<437::AID-HUMU9>3.0.CO;2-Q.
Direct sequencing analysis is largely used to confirm and characterize mutations previously detected by more rapid tests. We have developed a method-Comparative Sequence Analysis (CSA)-that simplifies the analysis of sequencing data facilitating its use as a first screen for mutation detection. Sequence data were split into their component electrophoretograms and the use of a size standard enabled equivalent traces from different individuals to be overlaid. This allowed simple and rapid visual analysis of the results. Using this technique in a blind study, we tested 576 samples for mutations in the Von Hippel-Lindau tumor suppressor gene, VHL. We were able to identify and characterize all 78 known mutations present within the sample set (100% sensitivity and specificity).
直接测序分析主要用于确认和表征先前通过更快速检测方法检测到的突变。我们开发了一种方法——比较序列分析（CSA），该方法简化了测序数据的分析，便于将其用作突变检测的初筛。序列数据被拆分为其组成的电泳图，使用大小标准可将来自不同个体的等效泳道叠加在一起。这使得结果能够进行简单快速的视觉分析。在一项盲法研究中，我们使用该技术对576个样本进行了冯·希佩尔-林道肿瘤抑制基因（VHL）的突变检测。我们能够识别并表征样本集中存在的所有78个已知突变（灵敏度和特异性均为100%）。
