Open Access

A novel RFLP‑ARMS TaqMan PCR‑based method for detecting the BRAF V600E mutation in melanoma

  • Authors:
    • Yunqing Zhang
    • Shoufang Qu
    • Jinyin Zhao
    • Ting Yu
    • Liping Guo
    • Songchao Yin
    • Xiaoxu Hu
    • Weijun Chen
    • Wei Lai
    • Jie Huang
  • View Affiliations

  • Published online on: May 30, 2018     https://doi.org/10.3892/ol.2018.8844
  • Pages: 1615-1621
  • Copyright: © Zhang et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )
Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )


Abstract

To enable the rapid and sensitive screening of the BRAF V600E mutation in clinical samples, a novel method combining restriction fragment length polymorphism (RFLP) analysis with the popular amplification refractory mutation system (ARMS) TaqMan quantitative (qPCR) genotyping method in a single reaction tube was developed. A total of 2 primer pairs were designed to enrich for and genotype the BRAF mutational hotspot (RFLP primers and ARMS primers) and a restriction enzyme was used to remove the wild‑type alleles. The analysis revealed that this method detected mutant alleles in mixed samples containing >0.1% mutant sequences. In a survey of 53 melanoma samples, this method detected 21 mutation‑positive samples. This novel RFLP‑ARMS TaqMan qPCR protocol may prove useful for detecting mutations in clinical samples containing only a small proportion of mutant alleles.
View Figures
View References

Related Articles

Journal Cover

August-2018
Volume 16 Issue 2

Print ISSN: 1792-1074
Online ISSN:1792-1082

Sign up for eToc alerts

Recommend to Library

Copy and paste a formatted citation
x
Spandidos Publications style
Zhang Y, Qu S, Zhao J, Yu T, Guo L, Yin S, Hu X, Chen W, Lai W, Huang J, Huang J, et al: A novel RFLP‑ARMS TaqMan PCR‑based method for detecting the BRAF V600E mutation in melanoma. Oncol Lett 16: 1615-1621, 2018
APA
Zhang, Y., Qu, S., Zhao, J., Yu, T., Guo, L., Yin, S. ... Huang, J. (2018). A novel RFLP‑ARMS TaqMan PCR‑based method for detecting the BRAF V600E mutation in melanoma. Oncology Letters, 16, 1615-1621. https://doi.org/10.3892/ol.2018.8844
MLA
Zhang, Y., Qu, S., Zhao, J., Yu, T., Guo, L., Yin, S., Hu, X., Chen, W., Lai, W., Huang, J."A novel RFLP‑ARMS TaqMan PCR‑based method for detecting the BRAF V600E mutation in melanoma". Oncology Letters 16.2 (2018): 1615-1621.
Chicago
Zhang, Y., Qu, S., Zhao, J., Yu, T., Guo, L., Yin, S., Hu, X., Chen, W., Lai, W., Huang, J."A novel RFLP‑ARMS TaqMan PCR‑based method for detecting the BRAF V600E mutation in melanoma". Oncology Letters 16, no. 2 (2018): 1615-1621. https://doi.org/10.3892/ol.2018.8844