Citation: Zishan Feng, Wei Zhang, Longjie Li, Bocheng Tu, Weicong Ye, Xiaofeng Tang, Hongbo Wang, Xianjin Xiao, Tongbo Wu. A cost-effective detection of low-abundance mutation with DNA three-way junction structure and lambda exonuclease[J]. Chinese Chemical Letters, ;2021, 32(2): 779-782. doi: 10.1016/j.cclet.2020.06.009 shu

A cost-effective detection of low-abundance mutation with DNA three-way junction structure and lambda exonuclease

Zishan Feng ^a ,
Wei Zhang ^{b, c} ,
Longjie Li ^a ,
Bocheng Tu ^a ,
Weicong Ye ^a ,
Xiaofeng Tang ^b ,
Hongbo Wang ^c ,
Xianjin Xiao ^{a, b, c, *,} ,
Tongbo Wu ^{a, b, *,}

a.
School of Pharmacy, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, China
b.
Institute of Reproductive Health, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, China
c.
Department of Obstetrics and Gynaecology, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430022, China

xiaoxianjin@hust.edu.cn

wutongbo@hust.edu.cn

Received Date: 10 May 2020
Revised Date: 28 May 2020
Accepted Date: 5 June 2020
Available Online: 17 June 2020

Figures(4)

We presented a low-abundance mutation detection method with lambda exonuclease and DNA three-way junction structure. The assistant strand in the DNA three-way junction structure could regulate the reaction system from the kinetics and thermodynamics aspects. The optimization of the assistant strand helps to improve the selectivity of the mutant-type DNA to the wild-type DNA about 35 times. Moreover, the cost of the optimization process could be saved by about 90%. The method was applied to the detection of a human ovarian cancer-related gene mutation BRCA1 (rs1799949, c.2082C>T). The limit of detection to the mutation abundance in the DNA three-way junction structure system (0.2%) was one order lower compared with that in the double-stranded DNA structure system (2%). The mutation abundance in different standard samples was quantitively measured, and the results were consistent with the initial abundance in the standard samples.
- Gene mutation,
- Cancer,
- Low abundance,
- DNA three-way junction structure,
- Lambda exonuclease
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