Abstract
In this article, a new kind of structure hairpin LNA probe was designed to detect BCR/ABL fusion gene in Chronic Myelocytic Leukemia. The LNA probe was immobilized on the gold electrode(AuE) through sulfur–Au interaction to construct specific electrochemical biosensor. The electrochemical response of the sensor to hybridization of the LNA probe with the target DNA was studied using [Cu(R)2]2+ as an electrochemical indicator. The optimal condition was discussed. The experimental results indicated that in pH 7.4 PBS buffer solution, this new method has excellent specificity for single base mismatch and complementary after hybridization. The relationship between the increased oxidation peak current of [Cu(R)2]2+ and the concentration of complementary strand was linear in the range of 1.0×10-8~1.0×10-6 mol/L. The detection limit was 2.0×10-9 mol/L.
Graphical Abstract
Keywords
hairpin LNA probe, electrochemical biosensor, BCR/ABL fusion gene, [Cu2(C7H5O2)4(C2H6O)2]
Publication Date
2011-05-28
Online Available Date
2011-05-06
Revised Date
2011-01-28
Received Date
2010-12-07
Recommended Citation
Hong-mei WANG, Li-qing LIN, Shao-huang WENG, Li-man WANG, Yin-huan LIU, Xin-hua LIN.
Hairpin Locked Nucleic Acids Probe Based Electrochemical Biosensor for Detection of BCR/ABL Fusion Gene[J]. Journal of Electrochemistry,
2011
,
17(2): 180-185.
DOI: 10.61558/2993-074X.2086
Available at:
https://jelectrochem.xmu.edu.cn/journal/vol17/iss2/6
References
[1] Zhang Li-Jun (张丽君), Zhang Rui(张蕊), Yu Jin-Xiang (于锦香)，et al. Analysis of BCR/ABL gene rearrangements and complex chromosomal aberrations in patients with chronic granulocytic leukemia [J]. Chinese Journal of Practical Internal Medicine(中国实用内科杂志), 2008, 28(2):130.
[2] WEI Na (魏娜), CHEN Jinghua (陈敬华), WANG Kun (王昆)，et al. Study on the Electrochemical Biosensor for Detection of PML /RARA Fusion Gene in Leukemia Based on Hairp in DNA Structure Probe [J]. Journal of Instrumental Analysis(分析测试学报), 2008, 27(9): 907.
[3] Zhong Zhi-Hong(钟志宏), Yuan Li-Ya (袁利亚), Chen Guo-An (陈国安). Effect of BCR-ABL Persistent Expression in Chronic Myelogenous Leukemia Blast Crisis [J].Journal of Oncology(肿瘤学杂志), 2006, 12(3): 245.
[4] Deng Meng-Feng (邓明凤), Wang Chang-Fu (王昌富), Zhang Wan-Sheng (张万胜),et al. Chromosome analysis and quantification of bcr-abl fusion gene transcripts in patients with chronic myeloid leukemia: Value of clinical application [J]. Chinese Journal of Clinical Laboratory Science（临床检验杂志）,2006, 24(3):179.
[5] Moravcová J, Rulcová J, Machová Poláková K, et al. Control genes in international standardization of real-time RT-PCR for BCR-AB [J]. Leukemia Research, 2009, 33(4):483.
[6] Xu Kai(许凯), Ye Zun-Zhong(叶尊忠), Ying Yi-Bin(应义斌). Electrochemical Deoxyribonucleic Acid Biosensor for Quantitative Detection of NOS Gene Sequence [J]. Chinese J Anal Chem（分析化学）, 2008, 36 (8):1113.
[7] Lin L Q, Chen J, Lin Q H, et al.. Electrochemical biosensor based on nanogold -modified poly-eriochrome black T film for BCR/ABL fusion gene assay by using hairpin LNA probe [J].Talanta, 2010, 80 (5): 2113.
[8] Wang Mao-Qing(王茂清), Du Xiao-Yan(杜晓燕), Liu Li-Yan(刘丽燕). Electrodeposited Pt-nanoparticles DNA biosensor for the detection of specific deoxyribonucleic acid in genetically modified soybean [J].Chinese J Anal Chem(分析化学), 2008, 36(7): 890.
[9] Obika S, Nanbu D, Hari Y, et al. Stability and structural features of the duplexes containing nucleoside analogues with a fixed N-type conformation, 2²-O,4²-C-methyleneribonucleosides [J]. Tetrahedron Lett., 1998, 39 (30): 5401.
[10] James Yang C Y, Wang L, Wu Y R, et al. Synthesis and investigation of deoxyribonucleic acid/locked nucleic acid chimeric molecular beacons [J]. Nucleic Acids Res, 2007, 35 (12): 4030.
[11] Koshkin A A, Rajwanshi V K, Wengel. Novel convenient syntheses of LNA [2.2.1]bicyclo nucleosides [J]. Tetrahedron Lett., 1998, 39(24): 4381.
[12] Braasch D A, Corey D R. Locked nucleic acid (LNA): fine-tuning the recognition of DNA and RNA [J]. Chem Biol, 2001, 8(1): 1.
[13] Wang J. DNA biosensors based on Peptide Nucleic Acid (PNA) recognition layers [J]. Biosens. Bioelectron, 1998, 13 (7/8 ): 757.
[14] Koshkin A A, Nielsen P, Meldgaard M, et al. LNA (Locked Nucleic Acid): An RNA Mimic Forming Exceedingly Stable LNA:LNA Duplexes [J]. J Am Chem Soc, 1998, 120 (50):13252.
[15] Fang S, Lee H J, Wark A W, et al. Attomole microarray detection of micro RNAs by nanoparticle-amplified SPR imaging measurements of surface polyadenylation reactions [J]. J Am Chem Soc, 2006, 128 (43): 14044.
[16] Lin L Q, Lin X H, Chen J H, et al. Electrochemical biosensor for detection of BCR/ABL fusion gene based on hairpin locked nucleic acids probe [J]. Electrochem Commun, 2009, 11 (8): 1650.
[17] Kang J, Huang X H, Xu J H, et al. Synthesis, crystal structure and antitumor activity in vitro of a new benzoate binuclear cupper(Ⅱ) Complex [J]. Chinese J Struct Chem, 2009, 28 (1) 15.
[18] Owczarzy R. Melting temperatures of nucleic acids: Discrepancies in analysis [J]. Biophys Chem,2005, 117 (3): 207.
[19] Dharuman V, Hahn J H. Effect of short chain alkane diluents on the label free electrochemical DNA hybridization discrimination at the HS-ssDNA/diluent binary mixed monolayer in presence of cationic intercalators [J]. Sens Actuators, B, 2007, 127 (2) 536.
