A型流感病毒流行毒株检测和分型基因芯片及使用方法

摘要

本发明公开一种A型流感病毒流行毒株检测和分型的基因芯片，它包括对H1N1、H3N2、H5N1、H9N2四种亚型进行检测和分型的79条特异性寡核苷酸探针、两条质控探针和载体，两条质控探针为质控探针I和质控探针II，其中特异性寡核苷酸探针和质控探针I分布在载体上，本发明还公开了基因芯片的制备方法，包括以下步骤：步骤一，探针的设计；步骤二，探针的合成；步骤三，芯片的制备。本发明公开基因芯片的使用方法，其特征是它包括以下步骤：步骤一，流感病毒RNA的提取；步骤二HA和NA基因PCR扩增；步骤三，荧光标记；步骤四，杂交；步骤五，杂交后清洗基因芯片；步骤六，扫描；步骤七，数据分析。

主权项

1、一种A型流感病毒流行毒株检测和分型的基因芯片，其特征是它包括对H1N1、H3N2、H5N1、H9N2四种亚型进行检测和分型的79条特异性寡核苷酸探针、两条质控探针和载体，两条质控探针为质控探针I和质控探针II，其中特异性寡核苷酸探针和质控探针I分布在载体上，对H1N1进行检测和分型的特异性寡核苷酸探针的序列编号 名称 序列1 H1N1-HA-1 atgccaacaactcaaccgac2 H1N1-HA-2 acttgagaagaatgtgacagtgac3 H1N1-HA-3 acacagaatgccattgacgg4 H1N1-HA-4 gaaattggaaatggctgctttga5 H1N1-HA-5 ctatatgagaaagtaagaagccagc6 H1N1-NA-1 ctagtgggagcagcatttcttt7 H1N1-NA-2 ttgtggcgttgatagtgatactg8 H1N1-NA-3 attcatctctttgttctatcagtgga9 H1N1-NA-4 atacacaaaagacaacagcataagaat10 H1N1-NA-5 ctaatggatggacagataccgatag11 H1N1-NA-6 gattggtcaggatacagtggaag对H3N2进行检测和分型的特异性寡核苷酸探针的序列12 H3N2-HA-1 ctgttacccttatgatgtgccg13 H3N2-HA-2 gaacgcagcaaagcctacag14 H3N2-HA-3 attaacagcacagggaatctaattg15 H3N2-HA-4 caataatgagatcagatgcaccca16 H3N2-HA-5 gaacaaagaagcaactgaggga17 H3N2-HA-6 gctgaggatatgggcaatgg18 H3N2-HA-7 atgacaacagcacggcaac19 H3N2-HA-8 caacaggtagaatatgcgacagt20 H3N2-HA-9 ctgttacccttatgatgtgccg21 H3N2-HA-10 ttatgcctcccttaggtcactag22 H3N2-HA-11 ggatgcggaatgtaccagaga23 H3N2-HA-12 aactagaggcatattcggcg24 H3N2-HA-13 gggtcaatcagaaatggaacttatg25 H3N2-NA-1 tgaccaacaccaccatagagaa26 H3N2-NA-2 agcagaatacagaaattggtcaaag27 H3N2-NA-3 atgcgatcctgacaagtgttatc28 H3N2-NA-4 ggacagggaacaacactaaacaa29 H3N2-NA-5 atggagtgaaaggctgggc30 H3N2-NA-6 tcctggtaactactgtaacattgc31 H3N2-NA-7 gccacaatatgcttccttatgc32 H3N2-NA-8 gtatctgaccaacaccaccatag33 H3N2-NA-9 ctgtgtgaaccaacaataatagaaa34 H3N2-NA-10 gctcaagttgtcacgatggaa35 H3N2-NA-11 cctattgatgaatgagttgggtgt36 H3N2-NA-12 gaggcttgtagatagtattggttca37 H3N2-NA-13 catagttgacagaggtaataggtcc对H5N1进行检测和分型的特异性寡核苷酸探针的序列38 H5N1-HA-1 cgacagagcaggttgacaca39 H5N1-HA-2 tcaagaaaggggactcagcaa40 H5N1-HA-3 aaggcaatagatggagtcacc41 H5N1-HA-4 gccgttggaagggaatttaataac42 H5N1-HA-5 aacaagaagatggaagacggatt43 H5N1-HA-6 cggaatggtcttacatagtggag44 H5N1-HA-7 ccagccaatgacctctgttac45 H5N1-HA-8 gctacaataataccaaccaagaaga46 H5N1-HA-9 cctaatgatgcggcagagc47 H5N1-HA-10 tgccattccacaacatacacc48 H5N1-HA-11 aagtgaattggaatatggtaactgc49 H5N1-HA-12 ccgcagtattcagaagaagcaa50 H5N1-HA-13 ttattcaacagtggcgagttcc51 H5N1-HA-14 aagaactgaaacacctattgagca52 H5N1-NA-1 tggaggaacaacatactgagaac53 H5N1-NA-2 ggcataataacagacactatcaaga54 H5N1-NA-3 agcactaattccaggagcgg55 H5N1-NA-4 ccagaactgacaggattagattgc56 H5N1-NA-5 attggtcaggatatagcgggag57 H5N1-NA-6 gccgttcatctcatgctcc58 H5N1-NA-7 ttgactcagggagccttgc59 H5N1-NA-8 tggaggaacaacatactgagaac60 H5N1-NA-9 ttactgtaatgactgatggaccaag61 H5N1-NA-10 ttcggagacaatccacgcc62 H5N1-NA-11 tggcatggctcaaatcggc63 H5N1-NA-12 agcactaattccaggagcgg64 H5N1-NA-13 ggtggactggaacggacag对H9N2进行检测和分型的特异性寡核苷酸探针的序列65 H9N2-HA-1 gcatcggctaccaatcaacaa66 H9N2-HA-2 tgttcctgtgacacatgcca67 H9N2-HA-3 atgccaaagaattactccacaca68 H9N2-HA-4 agaattgctagtgctgcttgaa69 H9N2-HA-5 caggtcagacattgcgagtg70 H9N2-HA-6 atggtatggacacattctttcagg71 H9N2-HA-7 acaaggactgacacaacaacaa72 H9N2-HA-8 aggtcagacattgcgagtaaga73 H9N2-HA-9 tctatggcaatccttcctgtga74 H9N2-HA-10 tcggattcattctacaggagca75 H9N2-HA-11 aataccacattgccattccaca76 H9N2-NA-1 ggtattctggtatcttctctgttga77 H9N2-NA-2 ccacggctaattccaagtcac78 H9N2-NA-3 aagtggcagaatacaagaattggt79 H9N2-NA-4 gtcaaattacagggttcgctcc两条质控探针的序列80 I cgtatataaaacggaacgtcgaagg81 II ccttcgacgttccgttttatatacg