applying microsatellite multiplex pcr analysis (mmpa) for determining allele copy-number status and percentage of normal cells within tumors应用微卫星多重pcr分析(mmpa)确定等位基因人类状态在肿瘤和正常细胞的比例.pdf

Applying Microsatellite Multiplex PCR Analysis (MMPA) for Determining Allele Copy-Number Status and Percentage of Normal Cells within Tumors 1 ´ 1 1 1 1 ´ 2 Carles Garcia-Linares , Jaume Mercade , Bernat Gel , Josep Biayna , Ernest Terribas , Conxi Lazaro , Eduard Serra1* ` ` 1 Institute of Predictive and Personalized Medicine of Cancer (IMPPC), Badalona, Barcelona, Spain, 2 Programa de Diagnostic Molecular de Cancer Hereditari, Laboratori de ` Recerca Translacional, Institut Catala d’Oncologia (ICO) – IDIBELL, L’Hospitalet de Llobregat, Barcelona, Spain Abstract The study of somatic genetic alterations in tumors contributes to the understanding and management of cancer. Genetic alterations, such us copy number or copy neutral changes, generate allelic imbalances (AIs) that can be determined using polymorphic markers. Here we report the development of a simple set of calculations for analyzing microsatellite multiplex PCR data from control-tumor pairs that allows us to obtain accurate information not only regarding the AI status of tumors, but also the percentage of tumor-infiltrating normal cells, the locus copy-number status and the mechanism involved in AI. We validated this new approach by re-analyzing a set of Neurofibromatosis type 1-associated dermal neurofibromas and comparing newly generated data with results obtained for the same tumors in a previous study using MLPA, Paralog Ratio Analysis and SNP-array techniques. Microsatellite multiplex PCR analysis (MMPA) should be particularly useful for anal