loading page

A tailored LNA clamping design principle: efficient, economized, specific and ultrasensitive for the detection of point mutations
  • +6
  • Hao Yang,
  • Mengqiu Yan,
  • Gaolian Xu,
  • Xiaohua Qian,
  • Ruiying Zhao,
  • Yuchen Han,
  • Ling Zhang,
  • Hongchen Gu,
  • Hong Xu
Mengqiu Yan
Author Profile
Gaolian Xu
Author Profile
Xiaohua Qian
Author Profile
Ruiying Zhao
Author Profile
Yuchen Han
Author Profile
Ling Zhang
Author Profile
Hongchen Gu
Shanghai Jiao Tong University
Author Profile
Hong Xu
Shanghai Jiao Tong University, Shanghai, China
Author Profile

Abstract

In the development of personalized medicine, the ultrasensitive detection of point mutations that correlate with diseases is important to improve the efficacy of treatment and guide clinical medication. In this study, locked nucleic acid (LNA) was introduced as an amplification suppressor of a massive number of wild-type alleles in an amplification refractory mutation system (ARMS) to achieve the detection of low-abundance mutations with high specificity and sensitivity of at least 0.1%. By integrating the length of clamp, base type, number and position of LNA modifications, we have established a “shortest length with the fewest LNA bases” principle from which each LNA base would play a key role in the affinity and the ability of single base discrimination could be improve. Finally, based on this LNA design guideline, a series of the most important single point mutation sites of epidermal growth factor receptor (EGFR) was verified to achieve the optimal amplification state which as low as 0.1% mutation gene amplification was not affected under the wild gene amplification was completely inhibited, demonstrating that the proposed design principle has good applicability and versatility and is of great significance for the detection of circulating tumor DNA.

Peer review status:UNDER REVIEW

02 May 2021Submitted to Biotechnology Journal
03 May 2021Assigned to Editor
03 May 2021Submission Checks Completed
17 May 2021Reviewer(s) Assigned