一种11个耳聋易感基因的PCR-LDR检测试剂盒

摘要

本发明提供了一种同时检测11个耳聋易感基因的PCR-LDR检测试剂盒，包括包装盒体(1)，PCR扩增和LDR反应试剂(2)-(5)；包括6对PCR引物、12组探针、1组通用探针和11组检测探针。该方法灵敏度高，判读清晰准确；相对传统测序技术，本试剂盒能完成一次性获得11个关键耳聋基因突变诊断结果的高通量筛查，具有经济、高效和高准确率等优势。

主权项

一种同时检测11个耳聋易感基因的PCR‑LDR检测试剂盒，其特征在于，所述试剂盒包括包装盒体(1)，PCR扩增和LDR反应试剂；所述PCR扩增试剂分别为PCR扩增Reaction Mix(2)和(3)；所述LDR反应试剂为LDR反应Probe Mix(4)和(5)；所述11个耳聋易感基因为GJB2：235delC、299‑230delA＞T、176‑191del16，SLC26A4：IVS7‑2A＞G、2168A＞G、1226G＞A、1174A＞T、1975G＞C、2027T＞A，线粒体12SrDNA：1555A＞G和1494C＞T；LDR通用模板：CCCATTAGTCTTGCGAGTACAGGTTTGCG通用荧光探针：P‑GAATCAATCAGACTG‑FAM   15其中用于GJB2基因三个位点的引物序列如下：GJB2_176_M：P‑TGGCTGCAGGGTGTTGCATTTTCGCAAACCTGTACTCGJB2_176del：TTTGCTAACTTCCCCTCTGACCCAGJB2_176_NOdel：TTTATCGTAGCACACGTTCTTGCAGCCGJB2_235_M：P‑GCTCGATAATGGGAAGTAGTTTTTTCGCAAACCTGTACTCGJB2_235_delC：TTTTTGCGTGGACACGAAGATCAGCTGCAGJB2_235_C：TTTTTTTGCGTGGACACGAAGATCAGCTGCAGGJB2_299_M：P‑ATGTGCGGTAGGAAACGTGTTTTTTTTTTCGTAAACGGTACTCGJB2_299_delAT：TTTTTTTTTTTGGTGCCTTTCCTCTTCTTCTCGJB2_299_AT：TTTTTTTTTTTTGACTATAGTCCTCTTCTTCTCATSLC26A4基因的6个检测位点引物序列如下：1975_M：P‑AAGGCGTTGCTTTGAAACTTTTCGCAAACGAGTTGTC1975_C：TAAAGATCATGCTCCAGTCTCACCAAG1975_G：TTTAAAGATCATCCTCCACCTTTCAGCTACAC2027_M：P‑GAGTTCTCACTAAGAACAACTTTTTTTCGCAAACCTCTAATC2027_T：TTTTTTAAAACCCTCAACACTTAGCCCCGCA2027_A：TTTTTTTTTAAAACTCAAAGGCTAACGGCT1174_M：P‑GCCGAACCCACTACGCATGAATTTTTTTTTCGCAAAATCAGCC1174_A：TTTTTTTCGGCAAAGAACTGAGCTCATT1174_T：TTTTTTTTTTCACCAGAATGATCCCTCAGAATCTG1226_M：P‑CCATCAAGAGCAGTACGTGACACTTTTTTTTTTTTTTCGCCTATGTGC1226_G：TTTTTTTTTCCTGTCTCCTGCACCTGGAGTTAC1226_A：TTTTTTTTTTTTCCGATGCGCTAGGCTGAACCTCTIVS7‑2_M：P‑GAAATCCGAACTTAATCATGTGAATTTTTTTTTTTTTTTTTTCGCAAATGTACTC56IVS7‑2_A：TTTTTTTTTTTGAAATCCCAGTAGCAATAATTGTCTIVS7‑2_G：TTTTTTTTTTTTTTGAAAGCAGTAGCCCTTATCCGACC2168_M：P‑GGCTGTCTAAACAATGTGTTTTTTTTTTTTTTTTTTTTTTCCTAAACCAGGACAC2168_A：TTTTTTTTTTTTTTTGGTTCCCATAGGTAGACTATAGGTCAT     2168_G： TTTTTTTTTTTTTTTTTTGGTTCTGTCGAATAATCTACCTCACTAC12s rRNA基因的2个检测位点的引物序列信息如下：1494_M：P‑CGATCGGGCGTCAGTGTCCCCGCTTTTTTTTTTTTCGCACCTCTGTACTC1494_C：TTTTTTTTTTTCCTTTGAGATCTATACTTCTTGAGG1494_T：TTTTTTTTTTTTTCCTTTGATCCTTATACAATGAGCCAACAA1555_M：P‑ATCATCCCTATAATCTGCGTATTTTTTTTTTTTTTTCGAATTCATCACTC1555_G：TTTTTTTTTTTTTTAGTTAACTTACCCAGTTACTGACTTAG1555_A：TTTTTTTTTTTTTTTTTAGCATCACGGTATAGATGTGCAACGACCCTA。