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题名: Isotachophoresis preconcentration integrated microfluidic chip for highly sensitive genotyping of the hepatitis B virus
作者: Liu, Dayu;  Shi, Ming;  Huang, Huaiqing;  Long, Zhicheng;  Zhou, Xiaomian;  Qin, Jianhua;  Lin, Bingcheng
通讯作者: 林炳承
关键词: microfluidic chip ;  sample preconcentration ;  isotachophoresis ;  hepatitis virus B ;  genotyping
刊名: JOURNAL OF CHROMATOGRAPHY B-ANALYTICAL TECHNOLOGIES IN THE BIOMEDICAL AND LIFE SCIENCES
发表日期: 2006-11-21
DOI: 10.1016/j.jchromb.2006.06.041
卷: 844, 期:1, 页:32-38
收录类别: SCI
文章类型: Article
部门归属: 18
项目归属: 1807
产权排名: 1;1
WOS标题词: Science & Technology ;  Life Sciences & Biomedicine ;  Physical Sciences
类目[WOS]: Biochemical Research Methods ;  Chemistry, Analytical
研究领域[WOS]: Biochemistry & Molecular Biology ;  Chemistry
英文摘要: The genotyping of hepatitis B virus (HBV) has become recently a valuable tool not only for epidemiological reasons but also for the clinical practice. Conventional methods for HBV genotyping typically include amplification of the target DNA sequences with a two-round nested PCR followed by separation of the amplified fragments by gel electrophoresis. A microfluidic chip that couples isotachophoresis (ITP) preconcentration and zone electrophoresis (ZE) separation may provide great advantages for sensitive, rapid and cost-effective clinical analysis. In this study, an HBV genotyping method with only one amplification round was developed by the application of the ITP-ZE chip. All the analysis steps of the ITP-ZE separation including sample injection, stacking and separation were performed continuously, controlled by sequential high-voltage switching. A 2.1 cm sample plug was preconcentrated between discontinuous buffers in ITP process, followed by ZE separation. Sensitivity enhancement was obtained through the increase of sample loading volume. The average LOD value of the ITP-ZE microfluidic chip was determined to be 0.0021 pg/mu L. In a large-scale HBV genotyping test, single round PCR products were analyzed by ITP-ZE microfluidic chip, and the results were compared with that of the conventional method. Among the 200 cases studied, the classification rate obtained with microfluidic chip was 93%, which was 6% higher than that obtained with the conventional method. Method with ITP-ZE chip analysis provides HBV genotyping information in reduced PCR amplification time with higher detection rate when compared with conventional method. This method holds great potential for extrapolation to the abundance of similar molecular biology-based techniques in clinical diagnosis. (c) 2006 Elsevier B.V. All rights reserved.
关键词[WOS]: CAPILLARY-ELECTROPHORESIS ;  SEPARATION ;  INFECTION ;  SAMPLE ;  PCR ;  FRAGMENTS ;  MICROCHIP ;  DEVICES ;  RISK
语种: 英语
原文出处: 查看原文
WOS记录号: WOS:000242328300005
Citation statistics: 
内容类型: 期刊论文
URI标识: http://cas-ir.dicp.ac.cn/handle/321008/98083
Appears in Collections:中国科学院大连化学物理研究所_期刊论文

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作者单位: 1.Chinese Acad Sci, Dalian Inst Chem Phys, Dept Biotechnol, Dalian 116023, Peoples R China
2.Dalian Univ Technol, Dept Biotechnol, Dalian 116023, Peoples R China
3.Guangzhou First Municipal Peoples Hosp, Dept Clin Lab, Guangzhou Med Coll, Guangzhou 510500, Peoples R China
4.Chinese Acad Sci, Grad Sch, Beijing 100000, Peoples R China

Recommended Citation:
Liu, Dayu,Shi, Ming,Huang, Huaiqing,et al. Isotachophoresis preconcentration integrated microfluidic chip for highly sensitive genotyping of the hepatitis B virus[J]. JOURNAL OF CHROMATOGRAPHY B-ANALYTICAL TECHNOLOGIES IN THE BIOMEDICAL AND LIFE SCIENCES,2006,844(1):32-38.
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