Establishment of pyrosequencing method to detect isocitrate dehydrogenase 1 mutations

doi:10.3969/j.issn.1006-7795.2014.02.016

Abstract

Abstract:

Objective The present study aimed to establish a pyrosequencing method for IDH1 mutation examination, and quantify the sensitivity of this method. Furthermore, we tried to compare the differences between the direct sequencing and pyrosequencing for IDH1 mutation examination. Methods Plasmids carrying either wide-type or mutant IDH1 gene were constructed, and used to optimize the pyrosequencing method. The exact amount of mutant plasmids mixed with wide-type plasmids were served as templates for the pyrosequencing reaction to quantify the sensitivity of pyrosequencing based mutation examination. Both direct sequencing and pyroquencing methods were used to detect IDH1 mutations in 96 gliomas. Results Pyrosequencing detected as low as 2 % IDH1 mutation mixed in wide-type gene. Among the 96 glioma samples examined, 32.3% of the samples were identified as carrying IDH1 mutations based on direct sequencing, while 74.0% based on pyrosequencing method. Conclusion Pyrosequencing is a reliable and sensitive method in detecting IDH1 mutation, which is suitable for molecular diagnosis.

Key words: isocitrate dehydrogenase 1, pyrosequencing, mutation

CLC Number:

Q754

Wang Danhui, Cai Yanning, Zhang Yanli, Gao Jie, Yang Caixia. Establishment of pyrosequencing method to detect isocitrate dehydrogenase 1 mutations[J]. Journal of Capital Medical University, 2014, 35(2): 219-224.

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URL: https://journal03.magtech.org.cn/Jweb_sdykdxxb/EN/10.3969/j.issn.1006-7795.2014.02.016

https://journal03.magtech.org.cn/Jweb_sdykdxxb/EN/Y2014/V35/I2/219

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