コバス® BRAF V600 変異検出キット
Contents
1. 5 000 ppm 0 5 0 5 12. DNA 5 000 ppm 0 59 DNase DNA
3. C 20 12 1 2 3 5 6 9 0 1 RXNMIX MGAC BRAF OM 15 30C 2 8
4. 2 3 5 DNA 5 000 ppm 0 5 DNA DNA 5 000 ppm 0 5 DNA 2
5. 2015 2015 1 3 1 2 22600AMX01329000 1 0 3 BRAF BRAF V600 2 4 5
6. BRAF V600 MUT Valid BRAF V600 WT Valid Mutation Detected Mutation Not Detected 3 No Mutation Detected Invalid 8 Failed 2 3 Mnatation Not Detected No Mutation Detected DNA 2 Invalid Failed Invalid
7. 9 V 256 1 257 15 161 176 271 162 433 256 271X100 94 5 161 162 100 99 4 417 433X100 96 3 9 10 V600E V600K V600D 11 V600E V600K V600D V600 1 NM 2 3 5 6 7 0
8. Invalid DNA DNA SD Invalid DNA DNA 207L A SD 20L DNA Invalid DNA 2 1 PCR 3 Invalid
9. Safe Lock microcentrifuge tube DNA TLB PK 1 5 mL 50 DNA 4 a DNA TLB PK 30 BRAF V600E2 FFPE DNA b 56 C 60 689 DNA 65 c 10 BRAF V600K V600D 2 d 90C
10. PCR BRAF 600 BRAF 1 BRAF V600 IV NO25026 BRAF V600 II IV 675 1000 mg m 3 1 960 mg 1 2 II 2010 12 30
11. DNA 5 ng L MGAC 7zL 28 35 42 49 56 63 70 77 84 91 xxL 100 125 150 175 200 225 250 275 300 325 DNA DNA lt 15 30 24 2 8 14 15 25 60 2 15 25 3 1 DNA 2 DNA DNA FFPE 1 125 ng DNA DNA 5 ng DNA Tissue Lysis Bufer DNA TLB 9 8 Proteinase K PK DNA Paraffin Binding Buffer DNA PBB DNA DNA Wash Buffer WBD DNA 5 ng gL 35 ng L DNA Wash Buffer II WB DD a DNA 7L DNA Elution Buffer DNA EB 35xL X 5ng zL
12. DNA PK 5 mL 4 5 mL 5 10 1 5 mL 15 mL 5 10 WB 1 15 30 90 50 mL 5 10 WBI 15 30C 90 50 mL WBI WBII 40 mL 37 450 20C 40 mL
13. 4 60 BRAF OM n 16 000 20 000 g 1 o 100x DNA Elution Buffer DNA BB RXNMIX BRAF OM MGAC RXNMIX 2 BRAF MUT BRAF WT 1 X 10L 5 8 000 g 1 DNA 5 mL DNA DNA BRAF OM 2 BRAF MUT BRAF WT 1 X 8zL
14. DNA BRAF 6 2 2 DNA 5 BRAF V600E FR FFPE V600K V600D BRAF V600 1 BRAF V600 RXNMIX 3x0 16 mL 2 5 dATP 2 5 dCTP 2
15. BRAF DNA Ct NaN 43 0 3 4 000 c mL BRAF H BRAF DNA Ct 31 5 38 5 BRAF V600 MUT 20 19 9 21 0 43 0 DNA Ct BRAF H 21 0 35 0 HH DNA Ct BRAF V600 MUT BRAF V600E DNA DNA DNA 10 BRAF V600 WT
16. 3 6 OS 0 37 95 0 26 0 55 Kaplan Meier 7 75 95 6 28 10 28 9 23 95 8 05 OS Log rank p lt 0 0001 PFS 0 26 95 0 20 0 33 Kaplan Meier 1 61 95 1 58 1 74 PFS 5 32 95 4 86 6 57 Log rank p lt 0 0001 4 I I 4
17. 105 0014 2 6 1 0120 600 152 105 0014 2 6 1 0120 600 152 PCR COBAS is a trademark of Roche Roche 25 L 25L PCR DNA 510 nm 465 nm DNA 580 nm 540 nm 6 6 0 7384505
18. DNA SD 5 6 1 5 mL RNase DNase DNA 5 10 7 DNA 1 z 480 5 10 3 z 480 2 1 74 mmol L 2 mg mL z 480 2 6 1 z 480 AD 25L 4
19. 5 dGTP 2 5 dUTP 2 5 dTTP Z05 DNA 2 BRAF V600 BRAF OM 3x0 13 mL TTS068 BRAF_F1 RL_BRAF_R5 DNA RAL_BRAF_MU23 3 BRAF V600 MGAC 3x0 15 mL 4 BRAF V600 MUT BRAF MUT 2x0 13 mL 5 BRAF V600 WT BRAF WT 2x0 13 mL 6 BRAF V600 CDNA SD 2x1 mL DNA BRAF V600E PCR FFPE DNA BRAF BRAF V600E BRAF 2
20. 2 6 2 BRAF MUT 25zL 2 3 BRAF WT 25zL 2 4 DNA 25 L 2 5 z 480 AD z 480 DNA 1 6 z 480 AD z 480 7 Review Accept 1 1 z 480
21. BRAF DNA DNA 1 V600E FFPE FFPE 2 DNA 6 HF ng PCR Rate 125 24 24 100 10 FFPET 8 1 62 5 24 24 100 31 25 24 24 100 125 24 23 96 5 24 24 100 596 FFPET 2 5 24 24 100 Blend 1 1 25 24 18 75 0 625 24 2 88 0 3125 24 17 71 125 24 24 100 5 24 22 92 5 FFPET 2 5 24 24 100 Blend 2 1 25 24 23 96 0 625 24 14 58 0 3125 24 12 50 125 24 24 100 5 24 24 100 596 FFPET 2 5 24 24 100 Blend 3 1 25 24 24 100 0 625 24 23 96 0 3125 24 17 71 2 5 125 24 0 0 FFPET 3 1 62 5 24 1 4 Blend 31 25 24 1 4 0 100 WT i 8 9 996 FFPE ED HE ng PCR Rate 125 48 48 100 15 63 48 48 100 7 81 48 47 98 1 6 1 3 9 48 4
22. 550 xL FFPE DNA DNA 00 8 000 g 1 DNA PCR kK 500xL DNA Wash Buffer 1 8 000 g 1 1 RXNMIX MGAC BRAF OM 15 30 C 500xL DNA Wash Buffer II 8 000 g RXNMIX MGAC BRAF OM 2 8 1 m
23. MGAC 2 BRAF MUT BRAF WT 1 X74 a DNA 5 Nanodrop VVis Sbeetrophotometer ND 1000 0F ND2000 1 DNA 0 1 EB BRAF MUT BRAF WT 1 20 ng L 1 2 13 415 6 7 8 9 10 10 20 ng L 2 ng L RXNMIX Ii0zL 40 50 60 7o so 9oliooliolizo iso BRAF OMI8SzL 32 40 48 56 64 72 80 88 96 104 2
24. 1 FFPE DNA 2 DNA PCR 4 E 1 6 cobas 1 2 AKAN E DNA 2 FFPE D 50 A
25. DNA 5 000 ppm 0 59 DNA 0 1 1 2 8C 16 Exp 5 000 ppm 5 6 BRAF V600 24 1
26. 60 1 V600R D594G G596R 1 3 1 5 mL K601E L597Q L597S 1 1 5 mL BRAF ARAF RAF1 5 RE 0 IO Ber AOE PCR 4 TRI 1 DNA 100 gL 3 h
27. 15 30 60 50 FFPE 50 DNA 1 DNA FFPE 2
28. DNA FFPE DNA DNA 0 5 709 DNA FFPE 1 HE DNA 5um 50 tr FFPE 12
29. 11 1 960 mg 1 2 2 1 I I 8 2 BRAF V600 JO028178 BRAF V600 75 096 95 34 9 96 8 5 RECIST ver1 1 CR PR 1 1 BRAF V600 MUT 2 BRAF DNA Ct 23 5 30 5 BRAF DNA Ct 23 5 29 0 2 BRAF V600 WT 2
30. 50 mL 5 40 mL 2 2 c 5 5 10 40 mL 50 mL 40 mL 2 2 DNA TLB 180 kg 5 mL 1 5 mL HE PK70zL DNA TLB P
31. 00 0 30 70 00 0 35 65 00 00 40 60 00 00 45 55 00 00 50 50 00 00 55 45 00 00 60 40 00 00 65 35 00 00 70 30 00 00 75 25 00 00 433 241 16 257 19 157 176 260 173 433 241 260X100 92 7 157 173 100 90 8 398 433X100 91 9 7 V600E V600K V600D 8 4 E V600E V600K V600D V600 35
32. 001 E
33. DNA ng D Fiter tubes with caps FT 5 DNA SD DNA zL Solection Tubes oD 35 mL DNA zL 5 NO DNA 21 ng L 0 0 Ah DNA L 35L X 5ng zL 21 ng 8 3 6 A SD L 35 8 3L 26 7L 5 mL 25 mL DNA 35 ng gz L 1 5 mL RNase DNase DNA 35 pr L DNA SD 37 DNA 5 xL 16 000 20 000 4 DNA uwL 5gL b DNA SD L 56 90 5L X DNA ng L 5 ng L 5L AO 2 2 z 480 A SD L 5L X 42 ng L 5ng L 5 37 3 3 DNA 4 1 5 mL
34. 7 98 1 95 48 39 81 0 98 48 34 71 125 48 48 100 7 81 48 48 100 3 9 48 48 100 2 11 5 1 95 48 47 98 0 98 48 47 98 0 49 48 45 94 125 48 47 98 31 25 48 47 98 15 63 48 41 85 3 4 4 7 81 48 43 90 3 9 48 43 90 1 95 48 32 67 DNA ng PCR Rate 125 60 58 97 31 25 60 60 100 15 63 60 57 95 7 81 60 59 98 1 4 5 3 90 60 57 95 1 95 60 49 82 0 98 60 47 78 0 49 60 46 77 6 Hit Rate X100 4 6 2 V600K V600D FFPE DNA FFPE DNA DNA DNA _ 0 eR Hit 1 V600 17 7 15 62 100 2 V600 21 5 125 0 3 3 V600 16 5 125 0 3 4 V600 22 71 125 0 3 5 V600 31 4 3 90 100 6 V600 34 6 3 90 100 7 V600 39 26 3 90 100 8 V600 36 07 1 95 100 9 V600 61 67 3 90 100 10 V600 68 75 0 49 100 DNA V600D V600K 5 95 0 0 10 90 00 15 85 00 a 20 80 00 25 75
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