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.
1. [1]
  STAMBOLIC V, SUZUKI A, DE LA POMPA J L, BROTHERS G M, MIRTSOS C, SASAKI T, RULAND J, PENNINGER J M, SIDEROVSKI D P, MAK T W. Cell, 1998, 95(1):29-39.
2. [2]
  DELGOFFE G M, KOLE T P, ZHENG Y, ZAREK P E, MATTHEWS K L, XIAO B, WORLEY P F, KOZMA S C, POWELL J D. Immunity, 2009, 30(6):832-844.
3. [3]
  CALDERON L, SCHINDLER K, MALIN S G, SCHEBESTA A, SUN Q, SCHWICKERT T, ALBERTI C, FISCHER M, JARITZ M, TAGOH H, EBERT A, MINNICH M, LISTON A, COCHELLA L, BUSSLINGER M. Sci. Immunol., 2021, 6(61):e5003.
4. [4]
  YANG M, SCHIAPPARELLI P, NGUYEN H N, IGARASHI A, ZHANG Q, ABBADI S, AMZEL L M, SESAKI H, QUINONES-HINOJOSA A, IIJIMA M. Oncogene, 2017, 36(26):3673-3685.
5. [5]
  XU W, ZHANG M J, LI Y, WANG Y, WANG K, CHEN Q Y, ZHANG R J, SONG W W, HUANG Q Q, ZHAO W H, WU J Q. Cancer Cell Int., 2021, 21(1):30.
6. [6]
  LI L H, YUE P L, SONG Q Q, YEN T T, ASAKA S, WANG T L, BEAVIS A L, FADER A N, JIAO Y C, YUAN G W, SHIH I M, SONG Y. J. Pathol., 2021, 253(1):119-128.
7. [7]
  FAGIN J A, WELLS S A. N. Engl. J. Med., 2016, 375(11):1054-1067.
8. [8]
  DAHIA P L, MARSH D J, ZHENG Z, ZEDENIUS J, KOMMINOTH P, FRISK T, WALLIN G, PARSONS R, LONGY M, LARSSON C, ENG C. Cancer Res., 1997, 57(21):4710-4713.
9. [9]
  LANDA I, POZDEYEV N, KORCH C, MARLOW L A, SMALLRIDGE R C, COPLAND J A, HENDERSON Y C, LAI S Y, CLAYMAN G L, ONODA N, TAN A C, GARCIA-RENDUELES M E R, KNAUF J A, HAUGEN B R, FAGIN J A, SCHWEPPE R E. Clin. Cancer Res., 2019, 25(10):3141-3151.
10. [10]
  DAVIES L, HOANG J K. Lancet Diabetes Endocrinol., 2021, 9(1):11-12.
11. [11]
  MIRANDA-FILHO A, LORTET-TIEULENT J, BRAY F, CAO B, FRANCESCHI S, VACCARELLA S, DAL MASO L. Lancet Diabetes Endocrinol., 2021, 9(4):225-234.
12. [12]
  FAN D, MA J, BELL A C, GROEN A H, OLSEN K S, LOK B H, LEEMAN J E, ANDERSON E, RIAZ N, MCBRIDE S, GANLY I, SHAHA A R, SHERMAN E J, TSAI C J, KANG J J, LEE N Y. Cancer, 2020, 126(2):444-452.
13. [13]
  DURANTE C, GRANI G, LAMARTINA L, FILETTI S, MANDEL S J, COOPER D S. JAMA, J. Am. Med. Assoc., 2018, 319(9):914-924.
14. [14]
  LIU M, RUAN M, CHEN L. Med. Oncol., 2014, 31(6):973.
15. [15]
16. [16]
  HUBER F, LANG H P, GLATZ K, RIMOLDI D, MEYER E, GERBER C. Nano Lett., 2016, 16(9):5373-5377.
17. [17]
  LI P, HE H, WANG Z, FENG M, JIN H, WU Y, ZHANG L, ZHANG L, TANG X. Anal. Chem., 2016, 88(1):883-889.
18. [18]
  TU M, CHIA D, WEI F, WONG D. Analyst, 2016, 141(2):393-402.
19. [19]
  ZAPPE K, PIRKER C, MIEDL H, SCHREIBER M, HEFFETER P, PFEILER G, HACKER S, HASLIK W, SPIEGL-KREINECKER S, CICHNA-MARKL M. Int. J. Mol. Sci., 2021, 22(22):12527.
20. [20]
  DU Y, POTHUKUCHY A, GOLLIHAR J D, NOURANI A, LI B, ELLINGTON A D. Angew. Chem., Int. Ed., 2017, 56(4):992-996
21. [21]
  WANG Y, TIAN K, SHI R, GU A, PENNELLA M, ALBERTS L, GATES K S, LI G, FAN H, WANG M X, GU L Q. ACS Sens., 2017, 2(7):975-981
22. [22]
  CHEN D, QI W, ZHANG P, ZHANG Y, LIU Y, GUAN H, WANG L. Pathology, 2018, 214(2):303-307
23. [23]
  MUKHERJEE S, SATHANOORI M, MA Z, ANDREATTA M, LENNON P A, WHEELER S R, PRESCOTT J L, COLDREN C, CASEY T, RIETZ H, FASIG K, WOODFORD R, HARTLEY T, SPENCE D, DONNELAN W, BERDEJA J, FLINN I, KOZYR N, BOUZYK M, CORRELL M, HO H, KRAVTSOV V, TUNNEL D, CHANDRA P. Canc. Genet., 2017, 216:128-141
24. [24]
  EMAUS M N, ANDERSON J L. Anal. Chim. Acta, 2020, 1124:184-193.
25. [25]
  BOTEZATU I V, KONDRATOVA V N, SHELEPOV V P, MAZURENKO N N, TSYGANOVA I V, SUSOVA O Y, LICHTENSTEIN A V. Anal. Biochem., 2020, 590:113517.
26. [26]
  NAKAMURA Y, TANIGUCHI H, IKEDA M, BANDO H, KATO K, MORIZANE C, ESAKI T, KOMATSU Y, KAWAMOTO Y, TAKAHASHI N, UENO M, KAGAWA Y, NISHINA T, KATO T, YAMAMOTO Y, FURUSE J, DENDA T, KAWAKAMI H, OKI E, NAKAJIMA T, NISHIDA N, YAMAGUCHI K, YASUI H, GOTO M, MATSUHASHI N, OHTSUBO K, YAMAZAKI K, TSUJI A, OKAMOTO W, TSUCHIHARA K, YAMANAKA T, MIKI I, SAKAMOTO Y, ICHIKI H, HATA M, YAMASHITA R, OHTSU A, ODEGAARD J I, YOSHINO T. Nat. Med., 2020, 26(12):1859-1864.
27. [27]
  LEBOFSKY R, DECRAENE C, BERNARD V, KAMAL M, BLIN A, LEROY Q, RIO FRIO T, PIERRON G, CALLENS C, BIECHE I, SALIOU A, MADIC J, ROULEAU E, BIDARD F C, LANTZ O, STERN M H, LE TOURNEAU C, PIERGA J Y. Mol. Oncol., 2015, 9(4):783-790.
28. [28]
  NEWTON C R, GRAHAM A, HEPTINSTALL L E, POWELL S J, SUMMERS C, KALSHEKER N, SMITH J C, MARKHAM A F. Nucleic Acids Res., 1989, 17(7):2503-2516.
29. [29]
  YU P C, TAN L C, ZHU X L, SHI X, CHERNIKOV R, SEMENOV A, ZHANG L, MA B, WANG Y, ZHOU X Y, JI Q H, WEI W J, WANG Y L. Endocr. Pract., 2021, 27(7):698-705.
30. [30]
  XU S, DUAN Y, LOU L, TANG F, SHOU J, WANG G. Oncol. Lett., 2016, 12(5):4238-4244.
31. [31]
  WEI J, ZHAO Z, GAO J X, WANG Y Q, MA L N, MENG X Y, WANG Z X. ACS Omega, 2020, 5(10):5365-5371.
32. [32]
  KOMIJANI M, SHAHIN K, AZHAR E I, BAHRAM M. Methods Mol. Biol., 2022, 2392:93-99.
33. [33]
  BRADLEY K M, ELMORE J B, BREYER J P, YASPAN B L, JESSEN J R, KNAPIK E W, SMITH J R. Genome Biol., 2007, 8(4):R55.
34. [34]
  BUTCHER L M, MEABURN E, LIU L, FERNANDES C, HILL L, AL-CHALABI A, PLOMIN R, SCHALKWYK L, CRAIG I W. Behav. Genet., 2004, 34(5):549-555.
1. [1]
  Ying Zhang , Fang Ge , Zhimin Luo . AI-Driven Biochemical Teaching Research: Predicting the Functional Effects of Gene Mutations. University Chemistry, 2025, 40(3): 277-284. doi: 10.12461/PKU.DXHX202412104
2. [2]
  Lin LI , Jiaxue LI , Meixia YANG , Jiayu DING , Jiaqi JING , Ruiping ZHANG . Preparation of mitoxantrone self-assembled carrier-free nanodrugs regulated by sodium acetate for apoptosis induction of human breast carcinoma cells. Chinese Journal of Inorganic Chemistry, 2025, 41(12): 2536-2548. doi: 10.11862/CJIC.20250138
3. [3]
  Heng Zhang . Determination of All Rate Constants in the Enzyme Catalyzed Reactions Based on Michaelis-Menten Mechanism. University Chemistry, 2024, 39(4): 395-400. doi: 10.3866/PKU.DXHX202310047
4. [4]
  Zhongxin YU , Wei SONG , Yang LIU , Yuxue DING , Fanhao MENG , Shuju WANG , Lixin YOU . Fluorescence sensing on chlortetracycline of a Zn-coordination polymer based on mixed ligands. Chinese Journal of Inorganic Chemistry, 2024, 40(12): 2415-2421. doi: 10.11862/CJIC.20240304
5. [5]
  Mi Wen , Baoshuo Jia , Yongqi Chai , Tong Wang , Jianbo Liu , Hailong Wu . Improvement of Fluorescence Quantitative Analysis Experiment: Simultaneous Determination of Rhodamine 6G and Rhodamine 123 in Food Using Chemometrics-Assisted Three-Dimensional Fluorescence Method. University Chemistry, 2025, 40(4): 390-398. doi: 10.12461/PKU.DXHX202405147
6. [6]
  Yu Dai , Xueting Sun , Haoyu Wu , Naizhu Li , Guoe Cheng , Xiaojin Zhang , Fan Xia . Determination of the Michaelis Constant for Gold Nanozyme-Catalyzed Decomposition of Hydrogen Peroxide. University Chemistry, 2025, 40(5): 351-356. doi: 10.12461/PKU.DXHX202407052
7. [7]
  Xiao SANG , Qi LIU , Jianping LANG . Synthesis, structure, and fluorescence properties of Zn(Ⅱ) coordination polymers containing tetra-alkenylpyridine ligands. Chinese Journal of Inorganic Chemistry, 2024, 40(11): 2124-2132. doi: 10.11862/CJIC.20240158
8. [8]
  Xudong Lv , Tao Shao , Junyan Liu , Meng Ye , Shengwei Liu . Paired Electrochemical CO₂ Reduction and HCHO Oxidation for the Cost-Effective Production of Value-Added Chemicals. Acta Physico-Chimica Sinica, 2024, 40(5): 2305028-0. doi: 10.3866/PKU.WHXB202305028
9. [9]
  Zeyi Yan , Ruitao Liu , Xinyu Qi , Yuxiang Zhang , Lulu Sun , Xiangyuan Li , Anchao Feng . Exploration of Suspension Polymerization: Preparation and Fluorescence Stability of Perovskite Polystyrene Microbeads. University Chemistry, 2025, 40(4): 72-79. doi: 10.12461/PKU.DXHX202405110
10. [10]
  Fanpeng Meng , Fei Zhao , Jingkai Lin , Jinsheng Zhao , Huayang Zhang , Shaobin Wang . Optimizing interfacial electric fields in carbon nitride nanosheet/spherical conjugated polymer S-scheme heterojunction for hydrogen evolution. Acta Physico-Chimica Sinica, 2025, 41(8): 100095-0. doi: 10.1016/j.actphy.2025.100095
11. [11]
  Wen-Bing Hu . Systematic Introduction of Polymer Chain Structures. University Chemistry, 2025, 40(4): 15-19. doi: 10.3866/PKU.DXHX202401014
12. [12]
  Quanliang Chen , Zhaohui Zhou . Research on the Active Site of Nitrogenase over Fifty Years. University Chemistry, 2024, 39(7): 287-293. doi: 10.3866/PKU.DXHX202310133
13. [13]
  Min Gu , Huiwen Xiong , Liling Liu , Jilie Kong , Xueen Fang . Rapid Quantitative Detection of Procalcitonin by Microfluidics: An Instrumental Analytical Chemistry Experiment. University Chemistry, 2024, 39(4): 87-93. doi: 10.3866/PKU.DXHX202310120
14. [14]
  Xue-Peng Zhang , Yuchi Long , Yushu Pan , Jiding Wang , Baoyu Bai , Rui Ding . 定量构效关系方法学习探索：以钴卟啉活化氧气为例. University Chemistry, 2025, 40(8): 345-359. doi: 10.12461/PKU.DXHX202410107
15. [15]
  Tengjiao Wang , Tian Cheng , Rongjun Liu , Zeyi Wang , Yuxuan Qiao , An Wang , Peng Li . Conductive Hydrogel-based Flexible Electronic System: Innovative Experimental Design in Flexible Electronics. University Chemistry, 2024, 39(4): 286-295. doi: 10.3866/PKU.DXHX202309094
16. [16]
  Jian Li , Yu Zhang , Rongrong Yan , Kaiyuan Sun , Xiaoqing Liu , Zishang Liang , Yinan Jiao , Hui Bu , Xin Chen , Jinjin Zhao , Jianlin Shi . Highly Efficient, Targeted, and Traceable Perovskite Nanocrystals for Photoelectrocatalytic Oncotherapy. Acta Physico-Chimica Sinica, 2025, 41(5): 100042-0. doi: 10.1016/j.actphy.2024.100042
17. [17]
  Xiaoli Sun , Xiang Wu , Li Gan , Wenming Wan . Barbier Polymerization: A New Teaching Case for Step-Growth Polymerization. University Chemistry, 2025, 40(4): 113-118. doi: 10.12461/PKU.DXHX202406102
18. [18]
  Yerong Chen , Bingbin Yang , Xinglei He , Yuqi Lin , Keyin Ye . Enzyme-Directed Evolution Enables Bioconversion of Organosilicon Compounds. University Chemistry, 2025, 40(10): 121-129. doi: 10.12461/PKU.DXHX202411054
19. [19]
  Ling Bai , Limin Lu , Xiaoqiang Wang , Dongping Wu , Yansha Gao . Exploration and Practice of Teaching Reforms in “Quantitative Analytical Chemistry” under the Perspective of New Agricultural Science. University Chemistry, 2024, 39(3): 158-166. doi: 10.3866/PKU.DXHX202308101
20. [20]
  Di WU , Ruimeng SHI , Zhaoyang WANG , Yuehua SHI , Fan YANG , Leyong ZENG . Construction of pH/photothermal dual-responsive delivery nanosystem for combination therapy of drug-resistant bladder cancer cell. Chinese Journal of Inorganic Chemistry, 2024, 40(9): 1679-1688. doi: 10.11862/CJIC.20240135

Get Citation

PDF

XML

Metrics

PDF Downloads(7)
Abstract views(639)
HTML views(34)

通讯作者: 陈斌, bchen63@163.com

1.
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

Metrics

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