Citation: WEI Jia, WANG Yao-Qi, WANG Zhen-Xin, MENG Xian-Ying. Highly Sensitive Detection of PTEN^R130* Mutation in Thyroid Cancer Based on Amplification Refractory Mutation System-Real Time Fluorescence-Quantitative Polymerase Chain Reaction[J]. Chinese Journal of Analytical Chemistry, ;2022, 50(5): 701-710. doi: 10.19756/j.issn.0253-3820.221008 shu

Highly Sensitive Detection of PTEN^R130* Mutation in Thyroid Cancer Based on Amplification Refractory Mutation System-Real Time Fluorescence-Quantitative Polymerase Chain Reaction

WEI Jia ^1,2 ,
WANG Yao-Qi ¹ ,
WANG Zhen-Xin ^2,, ,
MENG Xian-Ying ^1,,

1.
Department of Thyroid Surgery, The First Hospital of Jilin University, Changchun 130021, China;
2.
State Key Laboratory of Electroanalytical Chemistry, Changchun Institute of Applied Chemistry, Chinese Academy of Sciences, Changchun 130022, China

Corresponding author: WANG Zhen-Xin, MENG Xian-Ying,
Received Date: 4 January 2022
Revised Date: 18 February 2022

Fund Project: Supported by the Science and Technology Development Project of Jilin Province, China (No.20210204049YY) and the Shandong Taishan Industry Leading Talent Project, China (No.2019TSCYCX-24).

An analytical method based on amplification refractory mutation system-real time fluorescence-quantitative polymerase chain reaction (ARMS-qPCR) was developed for detection of gene mutation PTEN^R130* mutation in thyroid cancer. Allele-specific and reference primers for the PTEN^R130* mutation were designed for allele-specific and reference amplification of each sample at the same time, and the ΔCt value was obtained by Ct-allele-specific value minus Ct-reference value to analyze the variant allele frequency (VAF). The ARMS-qPCR method could detect VAF as low as 0.01%, with a linear range of 0.1% to 90% VAF, and could identify PTEN mutations in genomic DNA as low as 4 copies. This method successfully detected 5 cases of mutation with VAF>0.1% in 24 thyroid tumor samples. In addition, the ARMS-qPCR method also showed good detection performance for PTEN^R130* mutant spiked plasma samples under the electrochemical enrichment, indicating clinical practicability and the potential to detect trace PTEN^R130* mutation in complex samples.
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沈阳化工大学材料科学与工程学院沈阳 110142

Highly Sensitive Detection of PTENR130* Mutation in Thyroid Cancer Based on Amplification Refractory Mutation System-Real Time Fluorescence-Quantitative Polymerase Chain Reaction

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通讯作者: 陈斌, bchen63@163.com

Highly Sensitive Detection of PTEN^R130* Mutation in Thyroid Cancer Based on Amplification Refractory Mutation System-Real Time Fluorescence-Quantitative Polymerase Chain Reaction