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化学进展 2019, Vol. 31 Issue (6): 894-905 DOI: 10.7536/PC181008 前一篇   后一篇

所属专题： 电化学有机合成

• •

血液肿瘤相关生物标志物的电化学传感检测

宫苗1, 王晓英1,**(), 王晓宁2   

  1. 1.东南大学公共卫生学院 环境医学工程教育部重点实验室 南京 210009
    2.西安交通大学医学院第一附属医院血液内科 西安 710061
  • 收稿日期:2018-10-08 出版日期:2019-06-15 发布日期:2019-04-26
  • 通讯作者: 王晓英
  • 基金资助:
    国家自然科学基金项目(81302472); 国家自然科学基金项目(81600179); 江苏高校“青蓝工程”及江苏省研究生实践创新计划项目(SJCX18-0079)
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Electrochemical Sensing Detection of Biomarkers in Hematological Malignancies

Miao Gong1, Xiaoying Wang1,**(), Xiaoning Wang2   

  1. 1.Key Laboratory of Environmental Medicine Engineering of Ministry of Education, School of Public Health, Southeast University, Nanjing 210009, China
    2.Department of Hematology, the First Affiliated Hospital of Xi'an Jiaotong University, Xi'an 710061, China
  • Received:2018-10-08 Online:2019-06-15 Published:2019-04-26
  • Contact: Xiaoying Wang
  • About author:
    ** E-mail:
  • Supported by:
    National Natural Science Foundation of China(81302472); National Natural Science Foundation of China(81600179); the “Blue Project” Funded by Universities in Jiangsu, and the Postgraduate Practice Innovation Program of Jiangsu Province(SJCX18-0079)

血液肿瘤是一类严重危害人类健康的血液系统恶性疾病,主要累及骨髓、血液及淋巴组织。血液肿瘤相关生物标志物的定量检测是实现疾病精细化病理分型以实施靶向治疗进而改善预后的关键。本文对血液肿瘤相关生物标志物的种类、目前国内外常用的检测方法进行概述,重点阐述电化学生物传感器在检测血液肿瘤相关生物标志物方面的最新进展,并对血液肿瘤相关生物标志物电化学传感技术的未来发展方向提出展望,拟为其深入研究与应用提供参考。

关键词： 血液肿瘤, 生物标志物, 检测方法, 电化学生物传感器

Hematological malignancies(HM) is a kind of malignant disease of hematology system which seriously threatens human health, mainly involving bone marrow, blood and lymphatic tissue. The quantification of biomarkers in hematological malignancies is the key for fine stratification analysis, personalized targeted therapy and prognostic improvement. In this paper, the specific types and the commonly used detection methods at home and abroad of the hematological malignancies related biomarkers are summarized and compared. Specifically, the latest application of new electrochemical biosensor for the hematological malignancies related biomarkers is mainly described. Furthermore, the summary of its future directions and the potential applications is given, which provides reference for the further research and application of the biomarkers in hematological malignancies.

Key words: hematological malignancies, biomarker, detection methods, electrochemical biosensor
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表1 血液肿瘤中常见生物标志物[1,12~14]
Table 1 Common biomarkers in hematological malignancies
Hematological malignancies Main types Types of biomarker
DNA miRNA Protein Cell
Lymphoma NHL Proto-oncogene C-myc,Bcl-2, Bcl-6, Bcl-11 miR-21,
miR-155		 C-myc, Bax,
Bcl2,CD3/4/7/8/20/22/45		 Ramos
Tumor suppressor gene P53, P16
HL
Mutant gene B2M, PTPN1, TNFAIP3 - CD30/68/80 R-S
Leukemia AML Fusion gene PML/RARα,AML1-ETO,
CBFβ-MYH11		 miR-181a,
miR-15		 CD33 HL-60 /K562
ALL Mutant gene FLT3, PRAME, NPM1,
C-KIT, CEBPA		 miR-128/181/204/ 218/221/331 CD10,CD19 CCRF-CEM
CML Fusion gene P185BCR/ABL,TEL/AML1,
E2A/PBX1, MLL/AF4		 miR-217/221/222,
miR-17-92		 ABL K562
CLL Mutant gene PRAME miR-15/16/29/181 CD52 CLL
gene BCR/ABL, ASS
gene TP53, IGHV, ZAP70,TCL1
Multiple myeloma — MARK pathway KRAS, NRAS, BRAF miR-21/32/93/133
miR-206/221/222,
miR-17-92,		 CD30/38,
M protein,
CRP, LDH		 MM
NF-ΚB pathway TRAF3, CYLD, LTB
DNA repair pathway TP53, ATM, ATR
表2 血液肿瘤相关生物标志物常规检测方法
Table 2 Commonly used detection methods of the biomarkers in hematological malignancies
Method Detection Target Advantage Disadvantage ref
DNA FISH Translocation gene deletion/ amplification/rearrangement Bcl-2/6, IGH, C-myc, High specificity,
Less sample size		 Time-consuming,
Observed differences		 15
PCR Gene duplications/
deletions/mutations		 TP53, C-myc,
CDKN2A,
Bcl-2, REL		 High sensitivity,
Automation		 Complex operation procedure 16
DNA sequence Multiplegenes mutation,
copy-number changes,		 DNA High accuracy High cost,
Time-consuming		 17
miRNA RT-qPCR Single gene fusion,
amplifications/deletions		 BCR/ABL High sensitivity, Automation Sophisticated instrumentation 18
RNA sequence Multiple genesmutation,
structural variations		 RNA High accuracy High cost,
Time-consuming		 17
Protein IHC Positioning detection markers Bcl-2,CD3,
CD20,CD45		 High sensitivity Specialized technicians 19
ELISA Target antigen/antibody C-myc High selective low sensitivity, 20
Cell FCM Individual cell size,
theparticles within cells,		 BCR/ABL, P53,
CD19, CD20		 Microscale detection, consistency, repeatability Sophisticated instrumentation, qualifiedpersonnel 21
图1 标记型电化学和ECL检测BCR/ABL[28](A)和P16基因[33](B)
Fig. 1 Labeled electrochemical and ECL sensor for BCR/ABL fusion gene[28](A) and P16 gene[33](B)
表3 电化学方法检测血液肿瘤中DNA类生物标志物
Table 3 Electrochemical sensor for DNA biomarkers in hematological malignancies
Electrode Method Target Sensing strategy Linear range LOD ref
GCE DPV BCR/ABL AuNPs/PANI/CS-GS 10~1000 pmol·L-1 2.11 pmol·L-1 28
GCE DPV BCR/ABL CHA/GS/PANI/AuNPs 10 pmol·L-1~20 nmol·L-1 1.05 pmol·L-1 29
GE ASV BCR/ABL CSRP/QDs 10-14~10-10 mol·L-1 2 fmol·L-1 30
GCE ECL p16INK4a NPTh/RuAg@AuNPs 0.1 pmol·L-1~0.1 nmol·L-1 0.05 pmol·L-1 31
SPCE ECL p16INK4a PG/GR/CS/PPy/RuAg@AuNPs 0.1 pmol·L-1 ~0.1 nmol·L-1 0.05 pmol·L-1 32
GCE ECL CdkN2A/p16 PA6-MWCNTs-SiO2/RuAg@AuNPs 10-15~10-12 mol·L-1 0.5 fmol·L-1 33
GCE DPV BCR/ABL ISO bound to DNA 10 pmol·L-1~500 nmol·L-1 3 pmol·L-1 34
GCE DPV BCR/ABL Bio AuNPs/Gr/Pol 100 μmol·L-1 ~10 pmol·L-1 1.0 pmol·L-1 35
GE CA PML/RARα PCR/LNA 0.5~10 nmol·L-1 0.079 fmol·uL-1 36
SPCE SWV p185 BCR/ABL HRP/CNTs 8.3×10-14~1.7×109 mol·L-1 8.3 fmol·L-1 37
ITO CV BCR/ABL pDNA/CdTe/AS 10-12~10-6 mol·L-1 1.0 pmol·L-1 38
GE SWV FLT3 AMWNT/AuNPs 0.1 pmol·L-1 ~1 nmol·L-1 0.1 pmol·L-1 39
图2 标记型电化学方法检测miRNA-155[41](A)和C-myc蛋白[46](B)
Fig. 2 Labeled electrochemical sensor for miRNA-155[41] (A) and C-myc protein[46] (B)
表4 电化学方法检测血液肿瘤中RNA类生物标志物
Table 4 Electrochemical sensor for RNA biomarkers in hematological malignancies
Electrode Method Target Sensing strategy Linear range LOD ref
GCE SWV miR-21 Au-RGO/TiP-Cd2+/Ru(NH3)63+ 1.0 amol·L-1~10.0 pmol·L-1 0.76 amol·L-1 40
GCE CV miR-155 GO/Au/Cytc-p-TiO2@Pt/AOx@Pt 0.8 fmol·L-1~1 nmol·L-1 0.35 fmol·L-1 41
GE ASV miR-16 RCA/QDs 10-17~10-11 mol·L-1 0.32 amol·L-1 42
SPCE SWV miR-122 4 J-SENS 10 amol·L-1~1 fmol·L-1 2 amol·L-1 43
SPCE SWV miR-155 NCA/AuNPs - 3.57 fmol·L-1 44
图3 基于抗原和[48](A)和适配体识别的标记型电化学细胞传感器[50](B)
Fig. 3 Labeled electrochemical sensor based on antigen[48](A) and aptamer[50](B)
表5 标记型电化学方法检测血液肿瘤细胞
Table 5 Labeled electrochemical sensor for hematological malignancies cells
Electrode Method Target Identify element Linear range LOD ref
GCE DPV MCF-7 Bax antigen 2.5×103~1.6×105 cells·mL-1 800 cells·mL-1 48
GE DPV K562 T2-KK1B10 1.0×102~1.0×107 cells·mL-1 79 cells·mL-1 50
SPCE ECL K562 aptamer 1.0×102~2.0×107 cells·mL-1 46 cells·mL-1 51
GE DPV CCRF-CEM sgc8c 102~107 cells·mL-1 4 cells 52
SPE DPV CCRF-CEM sgc8c 10~106 cells·mL-1 10 cells·mL-1 53
GCE SWASV CCRF-CEM aptamer 1.0×102~1.0×106 cells·mL-1 10 cells 54
GE CV K562 aptamer 0~5.0×104 cells·mL-1 10 cells·mL-1 55
Au-PE DPV HL-60 aptamer 5.0×102~7.5×107 cells·mL-1 4 cells 56
GE LSV Ramos aptamer 10~106 cells 10 cells 57
GCE ECL Ramos TD05 1.0×102~1.0×104 cells·mL-1 55 cells·mL-1 58
GE ECL Ramos aptamer 20~500 cells 16 cells 59
GE ECL Ramos folic acid 150~9600 cells·mL-1 100 cells·mL-1 60
图4 无标记型电化学和ECL方法检测K562/ADM细胞 [61](A)和BCR/ABL基因[69](B)
Fig. 4 Label-free electrochemical and ECL sensor for K562/ADM cell[61] (A) and BCR/ABL gene[69] (B)
表6 无标记型电化学方法检测血液肿瘤中生物标志物
Table 6 Label-free electrochemical sensor for biomarkers in hematological malignancies
Electrode Method Target Sensing strategy Linear range LOD ref
GCE EIS K562/ADM AuNPs/PANI-NF/Ab 1.6×102~1.6×106 cells·mL-1 80 cells·mL-1 61
CPE EIS BCR/ABL PICA/ZnO/ssDNA 1.0×10-15~1.0×10-9 mol·L-1 2.2×10-16 mol·L-1 62
CPE EIS PML/RARA SPAN-GNO/pDNA 1.0×10-13~1.0×10-7 mol·L-1 3.2×10-14 mol·L-1 63
GE CV,EIS BCR/ABL AuNPs/PANI/ssDNA 10-8~10-15 mol·L-1 69.4 amol·L-1 64
GCE EIS MCF-7 PLL/AuNPs/Ab 5.0×102~1.6×106 cells·mL-1 170 cells·mL-1 65
GE ECL BCR/ABL Ru(phen)32+/N-HCR 10 fmol·L-1~1 nmol·L-1 5.49 fmol·L-1 69
CPE EIS BCR/ABL Fe3O4/CNTs/ssDNA 1.0×10-15~1.0×10-9 mol·L-1 2.1×10-16 mol·L-1 70
CPE EIS BCR/ABL FePt/GNO/ssDNA 1.0×10-14~1.0×10-9 mol·L-1 2.6×10-15 mol·L-1 71
GE EIS ZAP70 AuNPs/ssDNA 2.0×10-14~1.0×10-9 mol·L-1 4.0×10-15 mol·L-1 72
ITO EIS BCR/ABL Am-Si@ZnO/pDNA 1.0×10-6~1.0×10-16 mol·L-1 1×10-16 mol·L-1 73
NBR/PEDOT EIS NHL NBR/PEGDM/PEDOT fibre/PAA/ssDNA 1 amol·L-1~100 pmol·L-1 1×10-18 mol·L-1 74
GCE EIS K562 PLL/GO 1.0×102~1.0×107 cells·mL-1 30 cells·mL-1 75
图5 基于ECL和ASV分步检测靶DNA和癌细胞[67](A)和电化学同时检测Bcl-2和Bax蛋白[79](B)
Fig. 5 The detection of tDNA and cancer cells based on ECL and ASV[67](A)and simultaneous electrochemical detection of Bcl-2 and Bax proteins[79](B)
图6 基于纸基电极对肿瘤细胞的同时电化学传感检测[82]
Fig. 6 Paper-based electrodes for simultaneous electrochemical detection of multiple cancer cells[82]
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