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Development of a real-time RT-PCR assay for detecting EGFRvIII in glioblastoma samples.

Publication ,  Journal Article
Yoshimoto, K; Dang, J; Zhu, S; Nathanson, D; Huang, T; Dumont, R; Seligson, DB; Yong, WH; Xiong, Z; Rao, N; Winther, H; Chakravarti, A ...
Published in: Clin Cancer Res
January 15, 2008

PURPOSE: Epidermal growth factor receptor variant III (EGFRvIII) is an oncogenic, constitutively active mutant form of the EGFR that is commonly expressed in glioblastoma and is also detected in a number of epithelial cancers. EGFRvIII presents a unique antigenic target for anti-EGFRvIII vaccines and it has been shown to modulate response to EGFR kinase inhibitor therapy. Thus, detection in clinical samples may be warranted. Existing patents preclude the use of anti-EGFRvIII antibodies for clinical detection. Further, frozen tissue is not routinely available, particularly for patients treated in the community. Thus, detection of EGFRvIII in formalin-fixed paraffin-embedded (FFPE) clinical samples is a major challenge. EXPERIMENTAL DESIGN: We developed a real-time reverse transcription-PCR (RT-PCR) assay for detecting EGFRvIII in FFPE samples and analyzed 59 FFPE glioblastoma clinical samples with paired frozen tissue from the same surgical resection. We assessed EGFRvIII protein expression by immunohistochemistry using two distinct specific anti-EGFRvIII antibodies and examined EGFR gene amplification by fluorescence in situ hybridization. RESULTS: The FFPE RT-PCR assay detected EGFRvIII in 16 of 59 (27%) samples, exclusively in cases with EGFR amplification, consistent with the expected frequency of this alteration. The FFPE RT-PCR assay was more sensitive and specific for detecting EGFRvIII than either of the two antibodies alone, or in combination, with a sensitivity of 93% (95% confidence interval, 0.78-1.00) and a specificity of 98% (95% confidence interval, 0.93-1.00). CONCLUSION: This assay will facilitate accurate assessment of EGFRvIII in clinical samples and may aid in the development of strategies for stratifying patients for EGFRvIII-directed therapies.

Duke Scholars

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Published In

Clin Cancer Res

DOI

ISSN

1078-0432

Publication Date

January 15, 2008

Volume

14

Issue

2

Start / End Page

488 / 493

Location

United States

Related Subject Headings

  • Sensitivity and Specificity
  • Reverse Transcriptase Polymerase Chain Reaction
  • Paraffin Embedding
  • Oncology & Carcinogenesis
  • Humans
  • Glioblastoma
  • ErbB Receptors
  • 3211 Oncology and carcinogenesis
  • 3202 Clinical sciences
  • 1112 Oncology and Carcinogenesis
 

Citation

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Yoshimoto, K., Dang, J., Zhu, S., Nathanson, D., Huang, T., Dumont, R., … Mischel, P. S. (2008). Development of a real-time RT-PCR assay for detecting EGFRvIII in glioblastoma samples. Clin Cancer Res, 14(2), 488–493. https://doi.org/10.1158/1078-0432.CCR-07-1966
Yoshimoto, Koji, Julie Dang, Shaojun Zhu, David Nathanson, Tiffany Huang, Rebecca Dumont, David B. Seligson, et al. “Development of a real-time RT-PCR assay for detecting EGFRvIII in glioblastoma samples.Clin Cancer Res 14, no. 2 (January 15, 2008): 488–93. https://doi.org/10.1158/1078-0432.CCR-07-1966.
Yoshimoto K, Dang J, Zhu S, Nathanson D, Huang T, Dumont R, et al. Development of a real-time RT-PCR assay for detecting EGFRvIII in glioblastoma samples. Clin Cancer Res. 2008 Jan 15;14(2):488–93.
Yoshimoto, Koji, et al. “Development of a real-time RT-PCR assay for detecting EGFRvIII in glioblastoma samples.Clin Cancer Res, vol. 14, no. 2, Jan. 2008, pp. 488–93. Pubmed, doi:10.1158/1078-0432.CCR-07-1966.
Yoshimoto K, Dang J, Zhu S, Nathanson D, Huang T, Dumont R, Seligson DB, Yong WH, Xiong Z, Rao N, Winther H, Chakravarti A, Bigner DD, Mellinghoff IK, Horvath S, Cavenee WK, Cloughesy TF, Mischel PS. Development of a real-time RT-PCR assay for detecting EGFRvIII in glioblastoma samples. Clin Cancer Res. 2008 Jan 15;14(2):488–493.

Published In

Clin Cancer Res

DOI

ISSN

1078-0432

Publication Date

January 15, 2008

Volume

14

Issue

2

Start / End Page

488 / 493

Location

United States

Related Subject Headings

  • Sensitivity and Specificity
  • Reverse Transcriptase Polymerase Chain Reaction
  • Paraffin Embedding
  • Oncology & Carcinogenesis
  • Humans
  • Glioblastoma
  • ErbB Receptors
  • 3211 Oncology and carcinogenesis
  • 3202 Clinical sciences
  • 1112 Oncology and Carcinogenesis