Comparison of droplet digital PCR and conventional quantitative PCR for measuring EGFR gene mutation

Zhang,Bo; Xu,Chun‑Wei; Shao,Yun; Wang,Huai‑Tao; Wu,Yong‑Fang; Song,Ye‑Ying; Li,Xiao‑Bing; Zhang,Zhe; Wang,Wen‑Jing; Li,Li‑Qiong; Cai,Cong‑Li

doi:10.3892/etm.2015.2221

April-2015 Volume 9 Issue 4

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April-2015 Volume 9 Issue 4

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Article

Comparison of droplet digital PCR and conventional quantitative PCR for measuring EGFR gene mutation

Authors:
- Bo Zhang
- Chun‑Wei Xu
- Yun Shao
- Huai‑Tao Wang
- Yong‑Fang Wu
- Ye‑Ying Song
- Xiao‑Bing Li
- Zhe Zhang
- Wen‑Jing Wang
- Li‑Qiong Li
- Cong‑Li Cai
View Affiliations / Copyright

Affiliations: Department of Pathology, Affiliated Hospital of Academy of Military Medical Sciences, Beijing 100071, P.R. China, Wuhan YZY Medical Science and Technology Co., Ltd., Wuhan, Hubei 430075, P.R. China
Pages: 1383-1388
|
Published online on: January 27, 2015

https://doi.org/10.3892/etm.2015.2221
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Abstract

Early detection of epidermal growth factor receptor (EGFR) mutation, particularly EGFR T790M mutation, is of clinical significance. The aim of the present study was to compare the performances of amplification refractory mutation system‑based quantitative polymerase chain reaction (ARMS‑qPCR) and droplet digital polymerase chain reaction (ddPCR) approaches in the detection of EGFR mutation and explore the feasibility of using ddPCR in the detection of samples with low mutation rates. EGFR gene mutations in plasmid samples with different T790M mutation rates (0.1‑5%) and 10 clinical samples were detected using the ARMS‑qPCR and ddPCR approaches. The results demonstrated that the ARMS‑qPCR method stably detected the plasmid samples (6,000 copies) with 5 and 1% mutation rates, while the ddPCR approach reliably detected those with 5% (398 copies), 1% (57 copies), 0.5% (24 copies) and 0.1% (average 6 copies) mutation rates. For the 10 clinical samples, the results for nine samples by the ARMS‑qPCR and ddPCR methods were consistent; however, the sample N006, indicated to be EGFR wild‑type by ARMS‑qPCR, was revealed to have a clear EGFR T790M mutation with seven copies of mutant alleles in a background of 6,000 wild‑type copies using ddPCR technology. This study demonstrates the feasibility of applying the ddPCR system to detect EGFR mutation and identified the advantage of ddPCR in the detection of samples with a low EGFR mutation abundance, particularly the secondary EGFR T790M resistance mutation, which enables early diagnosis before acquired resistance to tyrosine kinase inhibitors becomes clinically detectable.

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Journals

International Journal of Molecular Medicine

International Journal of Oncology

Molecular Medicine Reports

Oncology Reports

Experimental and Therapeutic Medicine

Oncology Letters

Biomedical Reports

Molecular and Clinical Oncology

World Academy of Sciences Journal

International Journal of Functional Nutrition

International Journal of Epigenetics

Medicine International

Comparison of droplet digital PCR and conventional quantitative PCR for measuring EGFR gene mutation

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