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

Koji Yoshimoto, Julie Dang, Shaojun Zhu, David Nathanson, Tiffany Huang, Rebecca Dumont, David B. Seligson, William H. Yong, Zhenggang Xiong, Nagesh Rao, Henrik Winther, Arnab Chakravarti, Darell D. Bigner, Ingo K. Mellinghoff, Steve Horvath, Webster K. Cavenee, Timothy F. Cloughesy, Paul S. Mischel

研究成果: ジャーナルへの寄稿学術誌査読

80 被引用数 (Scopus)


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 bewarranted. 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.

ジャーナルClinical Cancer Research
出版ステータス出版済み - 1月 15 2008

!!!All Science Journal Classification (ASJC) codes

  • 腫瘍学
  • 癌研究


「Development of a real-time RT-PCR assay for detecting EGFRvIII in glioblastoma samples」の研究トピックを掘り下げます。これらがまとまってユニークなフィンガープリントを構成します。