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High specificity but low sensitivity of mutation-specific antibodies against EGFR mutations in non-small-cell lung cancer

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High specificity but low sensitivity of mutation-specific antibodies against EGFR mutations in non-small-cell lung cancer. / Bondgaard, Anna-Louise; Høgdall, Estrid; Mellemgaard, Anders; Skov, Birgit G.

I: Modern Pathology, Bind 27, Nr. 12, 12.2014, s. 1590–1598.

Publikation: Bidrag til tidsskriftTidsskriftartikelForskningfagfællebedømt

Harvard

Bondgaard, A-L, Høgdall, E, Mellemgaard, A & Skov, BG 2014, 'High specificity but low sensitivity of mutation-specific antibodies against EGFR mutations in non-small-cell lung cancer', Modern Pathology, bind 27, nr. 12, s. 1590–1598. https://doi.org/10.1038/modpathol.2014.67

APA

Bondgaard, A-L., Høgdall, E., Mellemgaard, A., & Skov, B. G. (2014). High specificity but low sensitivity of mutation-specific antibodies against EGFR mutations in non-small-cell lung cancer. Modern Pathology, 27(12), 1590–1598. https://doi.org/10.1038/modpathol.2014.67

Vancouver

Bondgaard A-L, Høgdall E, Mellemgaard A, Skov BG. High specificity but low sensitivity of mutation-specific antibodies against EGFR mutations in non-small-cell lung cancer. Modern Pathology. 2014 dec;27(12):1590–1598. https://doi.org/10.1038/modpathol.2014.67

Author

Bondgaard, Anna-Louise ; Høgdall, Estrid ; Mellemgaard, Anders ; Skov, Birgit G. / High specificity but low sensitivity of mutation-specific antibodies against EGFR mutations in non-small-cell lung cancer. I: Modern Pathology. 2014 ; Bind 27, Nr. 12. s. 1590–1598.

Bibtex

@article{3224606add89477a95bc82c9dffdea91,
title = "High specificity but low sensitivity of mutation-specific antibodies against EGFR mutations in non-small-cell lung cancer",
abstract = "Determination of epidermal growth factor receptor (EGFR) mutations has a pivotal impact on treatment of non-small-cell lung cancer (NSCLC). A standardized test has not yet been approved. So far, Sanger DNA sequencing has been widely used. Its rather low sensitivity has led to the development of more sensitive methods including real-time PCR (RT-PCR). Immunohistochemistry with mutation-specific antibodies might be a promising detection method. We evaluated 210 samples with NSCLC from an unselected Caucasian population. Extracted DNA was analyzed for EGFR mutations by RT-PCR (Therascreen EGFR PCR kit, Qiagen, UK; reference method). For immunohistochemistry, antibodies against exon19 deletions (clone 6B6), exon21 mutations (clone 43B2) from Cell Signaling Technology (Boston, USA) and EGFR variantIII (clone 218C9) from Dako (Copenhagen, DK) were applied. Protein expression was evaluated, and staining score (multipum of intensity (graded 0-3) and percentages (0-100%) of stained tumor cells) was calculated. Positivity was defined as staining score >0. Specificity of exon19 antibody was 98.8% (95% confidence interval=95.9-99.9%) and of exon21 antibody 97.8% (95% confidence interval=94.4-99.4%). Sensitivity of exon19 antibody was 63.2% (95% confidence interval=38.4-83.7%) and of exon21 antibody was 80.0% (95% confidence interval=44.4-97.5%). Seven exon19 and four exon21 mutations were false negatives (immunohistochemistry negative, RT-PCR positive). Two exon19 and three exon21 mutations were false positive (immunohistochemistry positive, RT-PCR negative). One false positive exon21 mutation had staining score 300. The EGFR variantIII antibody showed no correlation to EGFR mutation status determined by RT-PCR or to EGFR immunohistochemistry. High specificity of the mutation-specific antibodies was demonstrated. However, sensitivity was low, especially for exon19 deletions, and thus these antibodies cannot yet be used as screening method for EGFR mutations in NSCLC. Refinement of sensitivity for the mutation-specific antibodies is warranted to improve molecular diagnosis using EGFR immunohistochemistry.",
author = "Anna-Louise Bondgaard and Estrid H{\o}gdall and Anders Mellemgaard and Skov, {Birgit G}",
year = "2014",
month = dec,
doi = "10.1038/modpathol.2014.67",
language = "English",
volume = "27",
pages = "1590–1598",
journal = "Modern Pathology",
issn = "0893-3952",
publisher = "nature publishing group",
number = "12",

}

RIS

TY - JOUR

T1 - High specificity but low sensitivity of mutation-specific antibodies against EGFR mutations in non-small-cell lung cancer

AU - Bondgaard, Anna-Louise

AU - Høgdall, Estrid

AU - Mellemgaard, Anders

AU - Skov, Birgit G

PY - 2014/12

Y1 - 2014/12

N2 - Determination of epidermal growth factor receptor (EGFR) mutations has a pivotal impact on treatment of non-small-cell lung cancer (NSCLC). A standardized test has not yet been approved. So far, Sanger DNA sequencing has been widely used. Its rather low sensitivity has led to the development of more sensitive methods including real-time PCR (RT-PCR). Immunohistochemistry with mutation-specific antibodies might be a promising detection method. We evaluated 210 samples with NSCLC from an unselected Caucasian population. Extracted DNA was analyzed for EGFR mutations by RT-PCR (Therascreen EGFR PCR kit, Qiagen, UK; reference method). For immunohistochemistry, antibodies against exon19 deletions (clone 6B6), exon21 mutations (clone 43B2) from Cell Signaling Technology (Boston, USA) and EGFR variantIII (clone 218C9) from Dako (Copenhagen, DK) were applied. Protein expression was evaluated, and staining score (multipum of intensity (graded 0-3) and percentages (0-100%) of stained tumor cells) was calculated. Positivity was defined as staining score >0. Specificity of exon19 antibody was 98.8% (95% confidence interval=95.9-99.9%) and of exon21 antibody 97.8% (95% confidence interval=94.4-99.4%). Sensitivity of exon19 antibody was 63.2% (95% confidence interval=38.4-83.7%) and of exon21 antibody was 80.0% (95% confidence interval=44.4-97.5%). Seven exon19 and four exon21 mutations were false negatives (immunohistochemistry negative, RT-PCR positive). Two exon19 and three exon21 mutations were false positive (immunohistochemistry positive, RT-PCR negative). One false positive exon21 mutation had staining score 300. The EGFR variantIII antibody showed no correlation to EGFR mutation status determined by RT-PCR or to EGFR immunohistochemistry. High specificity of the mutation-specific antibodies was demonstrated. However, sensitivity was low, especially for exon19 deletions, and thus these antibodies cannot yet be used as screening method for EGFR mutations in NSCLC. Refinement of sensitivity for the mutation-specific antibodies is warranted to improve molecular diagnosis using EGFR immunohistochemistry.

AB - Determination of epidermal growth factor receptor (EGFR) mutations has a pivotal impact on treatment of non-small-cell lung cancer (NSCLC). A standardized test has not yet been approved. So far, Sanger DNA sequencing has been widely used. Its rather low sensitivity has led to the development of more sensitive methods including real-time PCR (RT-PCR). Immunohistochemistry with mutation-specific antibodies might be a promising detection method. We evaluated 210 samples with NSCLC from an unselected Caucasian population. Extracted DNA was analyzed for EGFR mutations by RT-PCR (Therascreen EGFR PCR kit, Qiagen, UK; reference method). For immunohistochemistry, antibodies against exon19 deletions (clone 6B6), exon21 mutations (clone 43B2) from Cell Signaling Technology (Boston, USA) and EGFR variantIII (clone 218C9) from Dako (Copenhagen, DK) were applied. Protein expression was evaluated, and staining score (multipum of intensity (graded 0-3) and percentages (0-100%) of stained tumor cells) was calculated. Positivity was defined as staining score >0. Specificity of exon19 antibody was 98.8% (95% confidence interval=95.9-99.9%) and of exon21 antibody 97.8% (95% confidence interval=94.4-99.4%). Sensitivity of exon19 antibody was 63.2% (95% confidence interval=38.4-83.7%) and of exon21 antibody was 80.0% (95% confidence interval=44.4-97.5%). Seven exon19 and four exon21 mutations were false negatives (immunohistochemistry negative, RT-PCR positive). Two exon19 and three exon21 mutations were false positive (immunohistochemistry positive, RT-PCR negative). One false positive exon21 mutation had staining score 300. The EGFR variantIII antibody showed no correlation to EGFR mutation status determined by RT-PCR or to EGFR immunohistochemistry. High specificity of the mutation-specific antibodies was demonstrated. However, sensitivity was low, especially for exon19 deletions, and thus these antibodies cannot yet be used as screening method for EGFR mutations in NSCLC. Refinement of sensitivity for the mutation-specific antibodies is warranted to improve molecular diagnosis using EGFR immunohistochemistry.

U2 - 10.1038/modpathol.2014.67

DO - 10.1038/modpathol.2014.67

M3 - Journal article

C2 - 24762545

VL - 27

SP - 1590

EP - 1598

JO - Modern Pathology

JF - Modern Pathology

SN - 0893-3952

IS - 12

ER -

ID: 138427908