EGFR突变检测引物/探针及方法(PCR/LDR法)

摘要

本发明公开了一组用于检测人类EGFR基因突变的引物、探针及使用方法，涉及肿瘤相关基因突变的检测。本发明的方法包括(1)提供引物和探针;(2)待测样品处理和模板提取;(3)配制改良的特异性PCR扩增突变基因序列的反应体系，以及检测基因突变的反应体系;(4)用方法(1)的引物和探针扩增富集待测的突变基因靶序列;(5)利用与扩增产物互补的荧光标记探针进行连接反应，ABI3130毛细管电泳条带大小作为结果的判断标准。本发明的引物、探针及方法:可同时检测EGFR基因的30多种点突变、插入缺失突变:灵敏度高:特异性强:选择性能力强:检测速度快，整个检测过程在2-3个小时内完成。

主权项

富集人类突变EGFR基因目标区及突变检测的引物和探针，引物和探针序列如下:SEQ IDSEQUENCESEQ NAME1CCTTCCAAATGAGCTGGCAAGTExon18F2CTCCCCACCAGACCATGAGAGExon18R3GCATGTGGCACCATCTCACAATExon19F4GAGAAAAGGTGGGCCTGAGGTTExon19R5AGCCACACTGACGTGCCTCTCExon20F6CTCCTTATCTCCCCTCCCCGTAExon20R7TTCATGCGCCTTTCCATTCTTTExon21F8GGTCCCTGGTGTCAGGAAAATGExon21R9CTCCGGTGCGTTCGGCACGGTGTATAAGTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTG71910TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTCTTGAAGGAAACTGAATTCAAAAAGATCAAAGTGCTGGCG719A11GCTCCGGTGCGTTCGGCACGGTGTATTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTG719‑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 TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTCGCAGCATGTCAAGATCACAGATTTTGGGCGL858R23GCTGGGTGCGGAAGAGAAAGAATACCATGCAGTTTTTTTTTTTTTTTTTTTTTTTTTTTTTL86124TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTCAAGATCACAGATTTTGGGCTGGCCAAACAL861Q25TGGTGCACCGCGACCTGGCAGCCTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTL83326TTTTTTTTTTTTTTTTTTTTTTTTTTTgGGCATGAACTACTTGGAGGACCGTCGCGL833V27TCAAGATCACAGATTTTGGL858R SelfX28CCACCGTGCAGCTCATCT790MSelfX29ATTTTGGGCTGGCCAL861QSelfX30CACTTTGATCTTTTTGAATTCAGTTE709G/ASelfX31TGCCGAACGCACCGG719ASelfX32CCGAACGCACCGGAGCG719S/CSelfX33ACTACTTGGAGGACCGTCL833VSelfX34TGGGGGTTGTCCACS768ISelfX。