luminex HPV
2002 501 3 3 3 HPV
103 HPV
2004 12.90/10 2005 10.16/10 2006 HPV
HPV HPV6 11 HPV16 18 CIN HPV 99.7%
HPV6 11 40 42 43 44 54 61 70 72 81 cp6 108 12 HPV16 18 31 33 35 39 45 51 52 56 58 59 68 73 82 15
HPV HPV
luminex HPV
HPV HPV
CIN
HPV DNA V-gene Animal Tissue Genomic DNA Mini-prep Kit DNA
HPV DNA MY09 5 -CGTCCMARRGGAWACTGATC-3 MY11 5 -GCMCAGGGWCATAAYAATGG-3 (M=A+C,R=A+G,W=A+T,Y=C+T)
94 5min 94 30s 54 30s 40 72 30s 72 5min
MIX Each PCR µl 10 PCR buffer 2.5 25mmol/LMgCl2 2.0 *10mmol/LdNTPs 0.5 25µmol/L MY-09 0.5 25µmol/L MY-11 0.5 Taq DNA 0.2 Template 2.0 ddh2o 16.8 Total 25.0
PCR 1.5g100µl1.5 10 µlpcr 6 DNA 100V 30min TANON GIS 2010
- PC03 5 -ACACAACTGTGTTCACTAGC-3 PC05 5 -GAAACCCAAGAGTCTTCTCT-3 DNAPCR 209bp
94 5min 94 30s 54 30s 40cycles 72 30s 72 5min MIX Each PCR µl 10 PCR buffer 2.5 25mmol/LMgCl2 2.0 *10mmol/LdNTPs 0.5 25µmol/L PC-03 0.5 25µmol/L PC-05 0.5 Taq DNA 0.2 Template 2.0 ddh2o 16.8 Total 25.0
HPV DNA SAP 1 ul ExonI 0.5ul PCR 3 ul 37 45min ddh2o 0.5ul 80 15min
CACTTGAACGGAAAGGAGTTAACCTCCCAAAAACTAAGGTTTTTATAG GGATCTGGCTTTTCCTTTTCAGGAGTGGGCTTTTGACAGGTAATGGCC TGTGACTGCACATACCTATAGGTATCTTCTAATGTACCATTTGGGGGA GGCGATAACCCAAAGTTCCAGTCTTCCAAAACAGAGGGATTCATTGTG TGAATATAGGCCATTACTTCAGCAGACAATGTAATGCTACATAATTGAA AAATAAATTGTAAATCATACTCTTCCACATGACGCATGTACTCTTTATAA TCAGAATTGGTGTATGTGGAAGATGTAGTTACGGATGCACATAATGTC ATGTTGGTACTGCGTGTGGTATCTACCACAGTAACAAACAGTTGATTAC CCCAACAAATACCATTGTTATGACCCTGGGCA
GNNNTCGACCGGGACNCCCNGTTNGNGGTNNTNANGGGGAAANGNAGAAGTN GGNTGNANTGGTCANANAAGGCGAGGGTTTCTGACANGNAAAGGCCTGTGACT GTNCATATCTATAAGTGTCNTCCAANGTACCATTTGGNGGAGGNGATAAACCAAA GTTCCAGNCCNCCAAAACAGAAGGATTCNTTGTGTGAATATAGGNCATGACTTCT GNAGATAATGTAANGCTACANAATTGAAAAACAAACTGTAAATCAAACTCCTCCA CATGGAGGATGTNCCCTTATAATCGGGAATTANCGNATGTCGAGANATCTAGAC ACAGATGCGCATANTGNCATATTTGTACTGCGCGTGGNATCTACCACAGTAACA NACNNNTGGTTTCNCCNGNAAATACCATTGTTATGACCCTGGGCAAGGANCCTC NTCTCTTNTACNNTTTCCTNTTTAACNAATNCTCCTNCTCCTNTTCTCCTNNCTNG TCNNGCTNNGCCNTCTNANNTCNATTNCACATCCCTCCNCTTTTTANCAGCTTCT TNNTTCCNCTATTCCTCCACGCGTGCCTTTTTCTNCNTCTTAGTACTATNCCTCN CANTACTCCTNCTCTTTNCTNCTNTNCANCCCCTTTCCTCNNATNCNNNNCTCTN NTTTNCATCNTCNTNNTCTNCTCTCCNTCCCANCNCCGTCNCNGACNGCCTTTN CCGCTCTCGNCCTTTGCANTTGNCTACTCANCNTNTNCNTANCCCTTTCTNNCTN CGCTANTNCCTACCNCTNCNTTTACCTNCCTNNACNNTTNGGCCCTTCNCNTTCC TCNCCTATCGCACTATAACGNGNANATCNTCCTCTTCNNTANACCNNNNNNNCC NACTNTCNNNCNAANCTCTCNCCTNNAGCC
HPV A 1 2 3 4 5 6 7 8 9 10 11 12 13 14 B 1 2 3 4 5 6 7 8 26 32 26 27 30 40 23 28 30 36 27 26 31 26 34 30 32 29 30 33 36 33 11 6 11 6 6 6 11 11 6 11 11 11 11 11 16 16 18/ 18/ 16/ 58 33 16/ CIN CIN CIN CIN CIN CIN CIN CIN
CA CIN HPV6 11 CIN HPV16 18 33 58 HPV
3
Luminex DNA PE HPV biotin HPV PE biotin
7 biotin 6 AGCCAGATCCCTATAAGAACCTTAG 11 ACAGGATCCCTATAAGGATATGAGTT 16 CACCTCCAGCACCTAAAGAAGAT 18 GGATGCTGCACCGGCTG 33 AGGAAGACCCCTTAGGTAAATATACAT 35 CCCAGTGCACCAAAACCTAAA 39 ATGCTCCAGCACCTGAAAAGA
94 5min 94 30s 54 30s 40cycles 72 30s 72 5min MIX Each PCR µl 10 PCR buffer 2.5 25mmol/LMgCl2 2.0 *10mmol/LdNTPs 0.5 (7 ) 2.0 Taq DNA 0.2 Template 2.0 ddh2o 16.8 Total 25.0
2µl PCR 3µl 4 SSC 50µl 55 30min 13 200r/min 5min 98µl 2 SSC 2µlPE 50 5min Bio-Plex
No Sequencing Luminex 6 11 16 18 33 35 39 A1 2 3 4 5 6 7 8 9 10 11 12 13 14 B1 2 3 4 5 6 si. 11 si. 6 si. 11 si. 6 si. 6 si. 6 si. 11 si. 11 si. 6 si. 11 si. 11 si. 11 si. 11 si. 11 si. 16 si. 16 mix. 18/? mix. 18/? mix. 16/? si. 33 41 438 30 222 42 56 50 1129 111 54 309 47 56 60 93 896 32 260 43 53 74 571 85 45 232 34 46 62 270 511 59 130 154 132 62 1314 124 30 78 31 35 15 71 1855 41 96 36 47 87 77 1380 54 123 43 44 71 528 137 36 136 40 33 39 936 2028 655 63 43 91 44 34 1462 65 78 55 49 70 42 693 67 92 64 58 39 120 1654 59 67 49 52 40 43 569 70 53 12 21 59 68 64 1271 116 23 81 77 88 75 373 275 75 107 81 312 80 57 323 41 75 39 312 80 57 323 41 75 49 60 1847 142 85 53 83 74 296 142 94 132 803 135 45
A B C A1 32 6 A2 27 11 A3 31 11 B1 34 16 B2 30 16 B3 33 58 C1 32 C2 25 C3 36
luminex luminex 100
luminex
luminex 100 35 60min 96 PCR
HPV DNA 15 7 HPV DNA CIN
HPV DNA HPV HPV DNA
HPV HPV luminex