HER2検査ガイド_h1_h4.ai
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1 1Hofmann M, et al. Assessment of a HER2 scoring system for gastric cancer:results from a validation study. Histopathology 2008;52: Lee AH, et al. The effect of delay in fixation on HER2 expression in invasive carcinoma of the breast assessed with immunohistochemistry and in situ hybridization. J Clin Pathol 2014;67: Rakha EA, et al. Updated UK Recommendations for HER2 assessment in breast cancer. J Clin Pathol 2015;68: Babic A, et al. The impact of pre-analytical processing on staining quality for H&E, dual hapten, dual color in situ hybridization and fluorescent in situ hybridization assays. Methods Dec;524: HER2. HER ,. 6Wolff AC, et al. Recommendations for human epidermal growth factor receptor 2 testing in breast cancer: American Society of Clinical Oncology/College of American Pathologists clinical practice guideline update. J Clin Oncol 2013 Nov 1;3131: HER2. HER ,. 8Laboratory General Checklist, CAP Accreditation Program, , College of American Pathologists HER2 I-VIEWHER24B5 ultraview HER24B5 Dual ISH HER AMX AMX AMX VENTANA 1062.TD.TMS.2000.HER01-003B
2 HER HER2HER2 HER2HER22001 T-DM1 HER2HER2 ToGA HER2 HER2HER2 2008Hofmann HER2 1 HER2 HER2 HER2 HER2 HER2I-VIEW HER24B5 ultraview HER24B5 Dual ISH HER2 IHC I-VIEW / ultraview HER24B5 Dual Color in situ HybridizationDISH Dual ISH HER2 HER2 HER2 I-VIEW / ultraview HER24B5 Dual ISH HER
3 IHC I-VIEWHER24B5 ultraview HER24B5Roche HER2IHC ultraview HER24B522300AMX HER24B HER24B5 5mL50 I-VIEWHER24B522100AMX ultraview DAB mL250 HER24B HER24B5 5mL50 -HRP DAB H2O2 IgG IgG/IgM 3,3' 25mL250 25mL250 25mL250 COPPER 25mL250 I-VIEW DAB : Ig -HRP DAB H2O2 IgG IgG/IgM 3,3' 25mL250 25mL250 25mL250 25mL250 25mL250 HER2HER2 HER2HER24B5 IgG DABH 2O 2COPPER HER2 HRP HRP DAB H2O2 HRP - HRP COPPER 25mL250 GX XT XT ULTRA LSAB HER2HER2 Ig -HRP HER2-HER24B5- IgG- DABH2O2COPPER - HRP HRP HRP DAB H2O2 HRP HRP Ig EZ 10X 2L / p.6 HER HI ULTRA 10X CC1 CC1 ULTRA 2L 2L 2L DS p / p HER2 4 in E-bar 1540 ULTRA 3 4
4 IHC EZ10 4.CC1 New Silane 4m 1mm5mm % mm 3 LOT 601 5SMEZ 1.EZ5 50mLEZ50mL+2.5g 150mLEZ150mL+7.5g 2.10 OK NG LOT SMEZ IMBR 1.DS41 25mL20mL+5mL 2. 3.Pretreatment24min
5 IHC HER2 4 in HER2 4 in Cell Line MDA-MB-231 T47D MDA-MB-453 BT-474 HER2 IHC <10% >10% >10% >10% HER2 HER Running Yes 1EZ 2CC1 3 I-VIEW DAB ultraview DAB BLOCKER A14 4BLOCKER B14 5Ig18 6HRP18 7DABH2O218 8COPPER I-VIEW DAB ultraview DAB Deparaffinization ULTRA Cell Conditioning Antibody AB Block Counterstain / CC1 Standard 37 / 32 min Hematoxylin 8 min / CC1 Standard 37 / 32 min Not Hematoxylin 8 min Post Counterstain Bluing Reagent 4min Bluing Reagent 4min 1.PCE-bar 2.WindowsVSS 3.Instrument View Ready p.6 7.5SMEZp
6 IHC HER2HER2 HER2 HER2HER2 HER2HER Heterogeneity % 10% equivocal 3.2~4 10~2040 / 10% / % HER2 0 HER2 10% / / 10% 10% / 10% / 10% equivocal HER2 HER2 Score 0 Score 1+ Score 0 Score 1+ Score 2+ Score 3+ Score 2+ Score
7 IHC Heterogeneity HER2 p.7 p.6 p.5 HER24B5 HER2 4m p.6 p.6 p.5 4m p.6 p
8 Dual Color HybridizationDISH Dual ISH HER2 Silver in situ Hybridization SISHDual Color in situ HybridizationDISH HER2HER217CEN17 Step1 Silver ISH for HER2 Black Signal Silver A Silver B Silver C Step2 RED ISH for Chromosome 17 Red Signal Goat anti-mouse Antibody AP ph Enhancer Naphthol Fast Red Mouse anti-dig Antibody 2 Dual ISH HER AMX Rabbit anti-dnp Antibody Goat anti-rabbit Antibody OH O O Dual ISH HER HER2 DNA 1 HER2 DNA HER2 2 CEN17 DNA CEN17 10mL50 17q11.2-q2 HO DNP NO2 NO2 HER2 DNA Probe DNP-labeled Chromosome17 Centromere Region DIG O H H OH H CEN17 DNA ProbeDIG-labeled HO ultraview SISH DNP DNP HRP A B C Ig 10mL100 10mL100 20mL100 10mL100 10mL100 HER217DNPHER2 DNA DIGChromosome 17 DNA DNPHRPDNPHER2 DNADNP HRPIg HRP ABCHER2 DIGAPDIGCEN17 DNA DIGAPIg APAS-TRNaphtholKLFast RedpH CEN17 FISH ultraview Red ISH DIG DIG AP ph Ig 10mL100 10mL100 20mL DISH FISH98.5% DISH FISH Naphthol Fast Red AS-TR KL 10mL100 20mL DISH FISH94.5% DISH FISH
9 Dual Color HybridizationDISH GX XT XT ULTRA XT XT / EZ 10X 2L p HI ULTRA 10X CC2 CC2 ULTRA SSC 10X ultraview Silver Wash 2L 2L 1L 2L 2L 83 DS p.17 p.19 p.20 p.15 20L3 CLSI ISH units/ml p HER2 Dual ISH 3-in E-bar 1540 ULTRA EZ10SSC5 4.CC2ultraView Silver Wash % DISHClinical and Laboratory Standards InstituteCLSI Clinical Laboratory Reagent WaterCLRW 8 UV mm 3 UV CLRW 10cfu/mL 10M/cm 0.22mTOC 500ppb 15 16
10 Dual Color HybridizationDISH New Silane HER2 4m HER2 1mm 5mm HER2 Dual ISH 3-in LOT 601 5SMEZ 1.EZ5 50mLEZ 50mL+2.5g 150mLEZ150mL+7.5g SMEZ IMBR 1.DS41 25mL20mL+5mL 2. OK NG xxxxxxxxxxx xxxxxxx xxxxxxx xxxxxxx xxxxxxxxx xxxxxxxxxxxxxxxx xxxx xxxxx xxxxxxxxx LOT HER2 Dual ISH 3-in Cell Line MCF 7 ZR-75-1 Calu-3 HER2 IHC Baking Deparaffinization Cell Conditioning ISH Protease Denaturation 1-2 copies/nuclei 3 copies/nuclei >6 copies/nuclei Hybridization Stringency Wash SISH Multimer SISH Chromogen Red Multimer RED Chromogen Counterstain Post Counterstain FISH HER2 Dual ISH 3-in-1 DISH ULTRA 63 / 20 min Depar CC2 / 86 Mild 8 min Standard12 min Extended8 min ISH Protease 3 8 min 20 min 6 hours min 4 min 24 min 8 min Hematoxylin - 8 min Bluing Reagent - 4 min / 3 / XT / GX CC2 Mild 8 min Standard12 min Extended8 min ISH Protease 3 8 min 20 min 6 hours min 4 min 24 min 8 min Hematoxylin - 8 min Bluing Reagent - 4 min >6 / 3.Pretreatment24min
11 Dual Color HybridizationDISH ULTRA 1.PCE-bar 2.WindowsVSS 3.Instrument View Ready 4.p p SMEZp Prime2 PrimeInstrument View Function Tests Test-Prime ULTRA Run Prime 11.PrimeRunning Yes 1EZ 2CC2 3ISH HER2 DNA 2 3DNP1 4HRP1 5A 2BC1 6DIG1 7AP1 8pH2Naphthol1Fast Red2 LCSEZ LCS Eukitt Entellan New Entellan HSR Malinol Cytoseal XYL Softmount Paramount DPX Cytoseal 60 Permount Histomount Ultramount Thermo EZ Mount SureMount Flo-texx Mountex Shandon Consul mount MM24 Pertex MicroMount Diamount Alcolmount BioMount 2 Acrytol Gel Mount Mount-Quick EMS Merck Merck Sysmex Muto Chemical Richard Allan Scientific WAKO Protaqs Quartett:Dako BDH:Raymond Lamb Richard Allan Scientific Fisher Raymond Lamb Dako Thermo Scientific Triangle Biomedical Sciences Lerner Labs Histolab Thermo Scientific SurgiPath Cell Path SurgiPath Diapath Diapath BBinternational SurgiPath Biomeda Daido Sangyo Co. Lemasol A Alcohol Aqueous Aqueous EZ dip30 p
12 Dual Color HybridizationDISH 3 93% 97% ULTRA Procedure : U Dual ISH CKT Baking Deparaffinization 63 / 20 min Extended Depara 63 / 20 min Depar A. 1ISH Protease 3 2Cell ConditioningCC2 B. 1ISH Protease 3 2Cell ConditioningCC2 Cell Conditioning ISH Protease CC2 / 86 Mild 12 min Standard12 min Extended12 min ISH Protease 3 16 min CC2 / 86 Mild 12 min Standard12 min Extended12 min ISH Protease 3 8 min C. Stringency Wash Denaturation Hybridization Stringency Wash SISH Multimer SISH Chromogen Red Multimer RED Chromogen Counterstain Post Counterstain 20 min 6 hours min 8 min 32 min 12 min Hematoxylin - 8 min Bluing Reagent - 4 min 20 min 6 hours min 4 min 32 min 12 min Hematoxylin - 8 min Bluing Reagent - 4 min A. B. C. Cell ConditioningCC min 1-3 cycles ISH Protease min Stringency Wash72-76 SISH Multimer12-60 min Silver Chromogen4-12 min Red Multimer12-60 min Red Chromogen4-12 min 21 22
13 Dual Color HybridizationDISH HER2CEN17CEN17 HER2HER2 HER2HER2 HER2HER2HER ~60HER2CEN HER2CEN HER2CEN17 2. HEIHC HER2Heterogeneity HER2 HER2Heterogeneity 17Polysomy 17Monosomy HER2 1 CEN171 HER2 2 CEN172 HER21 CEN HER CEN17 2 HER CEN172 60X HER2 1CEN17 1 HER2 4CEN17 2 HER2 CEN17 60XCEN17 HER2 2CEN
14 Dual Color HybridizationDISH 20HER2CEN1720CEN17HER2 HER2/CEN HER21 6.0ISHHER2/CEN172.0HER equivocalHER2/CEN172.0HER214.0 ISHequivocalIHC IHCISH IHC20CEN17HER2 HER2/CEN17HER2/CEN HER2/CEN HER2/CEN17 ISH 20HER2CEN17 HER2/CEN HER2/CEN17 <2.0 HER216.0 CEN17HER2 HER2/CEN17 20HER2/CEN HER2/CEN17 <2.0 HER214.0 <6.0 equivocal 20HER2/CEN17 <1.8>2.2 20HER2CEN17 12HER2/CEN17 HER2/CEN17 <2.0 HER21<4.0 HER2 40HER2/CEN17 < Heterogeneity HER2 HER
15 Dual Color HybridizationDISH p Dual ISH HER2 / p.22 p.16 p.22 p.16 4mp.17 4mp.17 p.17 p.19 p.19 p.20 p.19 p.17 p.22 p.17 p.19 p.20 p.17 p.16 4mp.17 p.22 p.16 4mp.17 DISH p.15 p.17 p.17 p.17 p.19 p.20 p.19 p.16 4mp.17 p.19 p.22 p
16 Dual Color HybridizationDISH
17 HER2 HER2HER2 56 HER2 HER2IHC2+ISHIHCISH HER2 HER2HER2 7 HER2IHC IHC IHC HER2 HER2ISHIHC >10 >10 // 10 >10 / 10 / IHC IHC0,1+,2+,3+IHC2+ISH IHC3+IHC2+ISHHER2/CEP172.0 IHC3 IHC2 equivocal IHC1 IHC0 ISHIHCISH IHC ISH HER2/CEP172.0 HER2/CEP17<2.0 ISH HER HER2 1 <4.0 HER HER <6.0 HER2 1 <4.0 HER2 ISH ISH ISH ISH equivocal ISH ISH IHCCh17ISH IHCISH Ch17 ISHHER2/CEP172.0HER2/CEP17<2.0 IHC ToGA FISHIHC3+5.1 HER2 HER
18 I-VIEWHER24B AMX Dual ISH HER AMX HER24B5 50 2~8 98, INFORM HER2 Dual ISH DNA Probe Cocktail Dual ISH HER2 50 2~8 400, Ventana iview DAB Universal Kit I-VIEW DAB g -HRP DAB H2O2 COPPER Endogenous Biotin Blocking Kit ~8 2~8 180,000 25, ultraview SISH DNP Detection Kit ultraview SISH DNP DNP HRP A B CultraView Red ISH DIG Detection Kit ultraview Red DIG DIG AP ph Naphtol Fast Red 100 2~8 450, ~8 450,000 ultraview HER24B AMX :50%25mL HybReady Solution 83 2~8 80, HER24B5 50 2~8 98, ISH Protease ISH units/ml ultraview Silver Wash ultraview Silver Wash 200 2~8 13,000 2L 15~30 28, VENTANA ultraview Universal DAB Detection Kit ultraview DAB -HRP DAB H2O2 COPPER 250 2~8 250, SSC SSC 10X CC 2 CC 2 2:5 3:XTGX 4:ULTRA Benchmark ULTRA CC 2 CC 2 ULTRA 2L 15~30 9, L 15~30 25, L 15~30 40, PATHWAY HER-2 4 in 1 Control Slides HER2 4 in ~8 88, HER2 Dual ISH 3-in-1 Xenograft Slides HER2 Dual ISH 3-in ~30 25, Rabbit Monoclonal Negative Control Ig 250 2~8 13,
VENTANA ALK D5F3 Rabbit Monoclonal Antibody OptiView ALK D5F3
VENTANA ALK D5F3 Rabbit Monoclonal Antibody OptiView ALK D5F3 2 OptiView ALK D5F3 11 10 79.3%31% 18% 80 85NSCLC 40% 1,2 1 1018 ALK ALK 2 ALK 3 5% ALK EML4 EML4-ALK Coild-Coil ALK ALK 3,4,5 2 6 EGFR 50%
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技術論文 乳癌における Dual Color in situ Hybridization 法に よる HER 遺伝子増幅の検出 FISH 法との比較 田中 京子 ) 武田麻衣子 ) 西川 武 ) 中井登紀子 ) 小関 久恵 ) 笠井 孝彦 ) ) 奈良県立医科大学附属病院病院病理部 6-85 ) 奈良県立医科大学病理診断学講座 ) 産業医科大学第 病理学教室 要 森田 剛平 ) 大林 千穂 ) 奈良県橿原市四条町
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