Evaluation of tumor microsatellite instability using five quasimonomorphic mononucleotide repeats and pentaplex PCR

微卫星不稳定性 微卫星 结直肠癌 多路复用 聚合酶链反应 生物 癌症 多重聚合酶链反应 等位基因 分子生物学 病理 遗传学 癌症研究 医学 基因
作者
Nirosha Suraweera,Alex Duval,Maryline Répérant,Chantal Vaury,Daniela Furlan,Karen Leroy,Raquel Seruca,Barry Iacopetta,Richard Hamelin
出处
期刊:Gastroenterology [Elsevier BV]
卷期号:123 (6): 1804-1811 被引量:531
标识
DOI:10.1053/gast.2002.37070
摘要

The microsatellite instability (MSI) phenotype is a characteristic of the hereditary nonpolyposis colorectal cancer syndrome as well as approximately 15% of sporadic colon and gastric tumors. It is a valuable diagnostic marker for the identification of hereditary nonpolyposis colorectal cancer cases and may be a molecular predictive marker for the identification of colon cancer patients who benefit from chemotherapy. To evaluate MSI, a reference panel was proposed at an international consensus meeting, comprised of 2 mononucleotide (BAT-25, BAT-26) and 3 dinucleotide repeats. Analysis of BAT-26 is sufficient for detecting the MSI phenotype in most, but not all, cases. Additional results with dinucleotide markers can sometimes lead to incorrect classification of MSI tumors.We describe here a single fluorescent multiplex system comprising 5 quasimonomorphic mononucleotide repeats for the detection of MSI tumors.None of 184 germline DNA samples, including 56 from African subjects, was found to contain allelic size variations in more than 2 of these markers. In contrast, all MSI tumors showed allelic size variations in 3 or more of the microsatellites. Using this assay, we confirmed (or reclassified in 6 cases) the MSI status of 124 colon and 50 gastric primary tumors and 16 colon cell lines.We propose that using a pentaplex polymerase chain reaction system allows accurate evaluation of tumor MSI status of DNA with 100% sensitivity and specificity without the need to match normal DNA. This assay is simpler to use than those involving dinucleotides and is more specific than using BAT-26 alone.
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