Table IIIa-CHER2 Measurement Methods

StudyAssays (Name)Criteria for PositivityTest Results (%)Comments
Adjuvant Chemotherapy
Yang et al. 2003 rec. # 8840 FISHnot doneFISHnot donePos36%HER2+ = IHC3+ by DAKO
scoring pre-ASCO/CAP
IHCNeomarker antibodyEquiv0
Neg64%
IHCstrong & complete membrane staining in >10% of tumor cells3+
2+
1+
0
Gusterson et al. 2003; rec. # 43690 FISHnot doneFISHnot donePos
IHCICR12 monoclonal antibodyEquiv
Neg
IHCstrong & complete membrane staining at dilution shown to give + signal if ≥3 copies of HER2 genePos16% of 760 node- pts; 19% of 746 node+ pts
Equivnone
Neg84% of 760 node- pts; 81 % of 746 node+ pts
Moliterni et. al. 2003; rec. # 10210

RCT; CMF (Grp1) vs CMF→A (Grp 2)
FISHnot doneFISHnot donePos
IHCCB11 antibodyEquiv
Neg
IHCstrong membrane staining found equivalent to 3+ by HercepTestPosGrp1:18.2%;Grp 2:16.2%
Neg75.6%73.3%
ND6.2%10.5%
Colozza et al. 2005; rec. # 3820 RCT; CMF (Grp 1) vs epirubicin (Grp 2); n=133 each tested for HER2 status FISHnot doneFISHnot done
IHCCB11 antibody and HercepTestIHC≥50% CB11+HER+Grp 1:28%Grp2:41%
≤50% CB11+HER-41%36%
CB11 negativeHER-31%23%
HercepTest using3+7%9%
DAKO scoring system2+7%9%
1+10%11%
075%71%
Pritchard et al. 2006; rec. #1760

RCT; CMF versus CEF;
FISHPathVysion kitFISHHER2/CEP17 ≥2.00Pos163 (26%)also reported concordance rates between the different assays used
IHCCB11 and TAB 250 antibodies (results reported separately from each antibody assay)Neg465 (74%)
PCRas described by O'Malley et al. 2001; rec. #13790)IHCcomplete membrane staining, score ≥5 on Allred semi-quantitative scaleCB11+124 (20%
CB11-510 (80%)PCR+ 195 (31%)
TAB250+116 (18%)PCR- 429 (69%)
TAB250-516 (82%)
Knoop et al. 2005; rec. # 3450

RCT (n=805 tested)
FISHpharmDxFISHHER2/CEP17 ≥2, as in kit manufacturer's manual; only tested if IHC2+ or 3+ followed instructions in manual for HercepTest kit PosIHC2+: 21.0IHC3+: 89.4
IHCHercepTestEquiv6.28.9
Neg72.81.6
IHC3+30.6
2+10.1(IHC3+ or FISH+) = HER2+: 32.7%
1+32.7HER2-: 67.3%
026.7
Dressler et al. 2005, rec. # 4280; Thor et al. 1998, rec. # 40880 FISHPathVysion kitRCT arm:hiqh-dosemod-doselow-dosetotal
IHCCB11 (n=346) or A0-11-854 (n=177) antibodiesFISHHER2/CEP17 >2Pos30 (5.7%)31 (5.9%)30 (5.7%)91 (17%)
Neg149 (28.4%)136 (26.0%)148 (28.2%)433 (83%)
PCRdifferential PCR assay as described in Thor et al. 1998IHC≥50% of invasive cells stained by antibodyPos44 (8.4%)43 (8.2%)40 (7.7%)127 (24%)
Neg134 (25.6%)124 (23.7%)138 (26.4%)396 (76%)
PCR“unequivocal amplification relative to ...normal...and amplified standard controls”Pos30 (6.1%)31 (6.3%)30 (6.1%)91 (18%)
Neg131 (26.7%)125 (25.5%)144 (29.3%)400 (82%)
Del Mastro et al. 2004, 2005; rec. # 48020 FISHnot doneFISHnot donePos
IHCCB11 antibodyNeg
FEC14 (n=370)FEC21 (n=361)
all slides scored by one pathologist, blinded to treatment arm & outcomeIHC3+ score on Dako scale: 103 of 731 (14%) with specimens available for assay3+50 (13.5%)53 (14.7%)
2+24 (6.5%)23 (6.4%)
1+19 (5.1%)20 (5.5%)
0277 (74.9%)265 (73.4%)
Tanner et al. 2006; rec. # 1820 data for n=180 from FEC arm tested for HER2 status FISHnot doneCISH≥6 copies in >30% of invasive carcinoma cells or ratio >2, HER2/CEP17Pos56 (31%)
CISHZymed probes (digoxigenin-labeled)Equiv0
IHCnot doneNeg124 (69%)
IHC3+
2+
1+
0
Hayes et al. 2007; rec. # 47610 FISHPathVysion kitFISHHER2/CEP17 ≥2.00Posproportions of HER2+ and HER2- patients not reported for any assay method
IHCCB11 antibody and HercepTestEquiv
Neg
IHCCB11: HER2+if ≥50% of breast cancer cells were stained; Herceptest: as in Dako manual3+
2+
1+
0
Martin et al. 2005; rec # 47650 FISHnot reportedFISHHER2/CEP17 ≥2.00Pos319 (21.4%; 20.8%, DAC arm; 22.0% FAC arm)
IHCCB11 antibody (only for 12 patients)Neg943 (63.3%; 63.8%, DAC arm; 62.7% FAC arm)
???229 (15.4%; 15.4%, DAC arm; 15.3% FAC arm)
IHCnot reported
Neoadjuvant (Pre-operative) Chemotherapy
Learn et al. 2005; rec. # 47640

n=104 classified for HER2 status
FISHnot reportedFISHnot reported
IHCTAB250 antibody (Zymed; South San Francisco, CA)HER2 Pos41 (39.4% of those tested)
FISH performed on all specimens with “borderline” HER2 IHC scoresIHCnot reportedHER2 Neg63 (60.6% of those tested)
Arriola et al. 2006; rec # 950

n=223 tested by IHC/FISH initial algorithm & by CISH
FISHOncor/Ventana Inform kitCISH>5 copies or ratio>2 for>5 copiesratio>2
CISHZymed probe and Spot-Light kitHER2/CEN17Pos18%14%
IHCCB11 antibody and HercepTestNeg82%86%
algorithm: CB11 first, then HercepTest for negatives only, then FISH for discordant IHC results; positives by initial algorithm tested byCISHIHC/FISH initial algorithm: CB11+ if complete membrane staining in >10% of cells; HercepTest+ if 2+ or 3+; FISH+ if >4 signals/cellPos19%
Neg 81%
Park et al. 2003; rec # 9960 FISHnot doneFISHnot done
CISHZymed SPOT-Light HER2 probe, digoxigenin-labeledCISHHER2 gene copy # >4, or large gene copy cluster in >50% of cancer cell nudePos46%
Neg54%
IHCnot doneIHCnot done
Zhang et al. 2003; rec # 9820 FISHPathVysion kitFISHgene copy ratio >2.0, HER2/chromosome 17 centromerePos13%overall, 28% (n=28) HER2+ defined as 3+ by IHC or FISH+; 72% (n=69) HER2-
IHCAB8 Neomarker antibodyNeg36%
untested51%
n=75 analyzed by IHCIHCstrong membrane staining in ≥10% of tumor cells3+23%
n=48 analyzed by FISH2+9%
n=97 all patients in study1+10%
035%
untested23%
Tulbah et al. 2002; rec # 11560

n=54 tested
FISHnot doneFISHnot donePos
IHCHercepTestEquiv
Neg
IHCscored 0–3+, as in Dako kit guide; for analysis of response, only 3+ was considered HER2+3+22 (41%)
2+12 (22%)
1+8 (15%)
012 (22%)
Tinari et al. 2006; rec # 2300

retrospective single-arm series; n=77
FISHnot reportedFISHnot reported; used only if HercepTest scored 2+Pos20 (26%)
IHCHercepTest (Dako)Equiv0
Neg57 (74%)
IHCscored 0–3+, as in Dako kit guide; positive if IHC scored 3+ or if FISH+ and IHC 2+3+
2+
1+
0
Chemotherapy for Advanced or Metastatic Disease
Harris et al 2006; rec. # 390 FISHVysis PathVysion kit (Vysis Inc, Downers Grove, IL)FISHRatio of HER2 to CEP17 signal ≥ 2.0.Pos26Cohen's kappa = 83.0%
(SE5.3%) for FISH vs CB11;
72.0%(SE6.2%) for Hercep-Test (0–1 vs 2–3) vs FISH;
79.2%(SE6.0%) for Hercep-Test (0–2 vs 3) vs FISH;
70.0%(SE6.3%) for Hercep-Test (0–1 vs2–3) vs CB11;
84.2%(SE5.4%) for Hercep-Test (1–2 vs 3) vs CB11.
Neg74
IHCMonoclonol antibody CB11 (Biogenex, San Ramon, CA); HercepTest (Dako Corp, Carpinteria, CA)IHCCB11 : moderate to strong intensity staining in ≥10% of invasive carcinoma cells.
Pos20
Equiv
Neg80
Herceptest score of 3+; i.e., complete membrane staining of >10% tumor cells040
128By CB11, 9% of African American women are HER2+ vs 20% of Caucasian women (p=0.08).
211
321
Di Leo et al 2004; rec. # 5970 IHCCB-11 (Novocastra, Newcastle, UK)Grp1Grp2
IHC →FISH: FISH done if IHC stained membranes in ≥ 1% of invasive cells; HER2+ if signal ratio, HER2/CEP17 ≥2Pos16%25%
FISHSpectrum Orange HER-2/Spectrum Green CEP17 (PathVysion, Vysis, Downers Grove, IL)unknown14%16%
Neg69%59%
Konecny et al 2004; rec. # 6740FISHPathVision HER-2 Neu & CEP17 probes (Vysis, Downers Grove, IL)Grp1Grp2
FISH≥2 HER-2/neu genes per Chromosome 17 CentromerePos35.834.8
Equiv
Neg64.265.2
IHCnot done

From: Appendix C. Evidence Data Abstraction Tables

Cover of HER2 Testing to Manage Patients With Breast Cancer or Other Solid Tumors
HER2 Testing to Manage Patients With Breast Cancer or Other Solid Tumors.
Evidence Reports/Technology Assessments, No. 172.
Seidenfeld J, Samson DJ, Rothenberg BM, et al.

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