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学科主题: 临床医学
题名:
A fully sealed plastic chip for multiplex PCR and its application in bacteria identification
作者: Xu, Youchun1; Yan, He1; Zhang, Yan2,3; Jiang, Kewei4; Lu, Ying1,3; Ren, Yonghong5; Wang, Hui4; Wang, Shan4; Xing, Wanli1,2,3
刊名: LAB ON A CHIP
发表日期: 2015
DOI: 10.1039/c5lc00244c
卷: 15, 期:13, 页:2826-2834
收录类别: SCI
文章类型: Article
WOS标题词: Science & Technology
类目[WOS]: Biochemical Research Methods ; Chemistry, Multidisciplinary ; Nanoscience & Nanotechnology
研究领域[WOS]: Biochemistry & Molecular Biology ; Chemistry ; Science & Technology - Other Topics
关键词[WOS]: POINT-OF-CARE ; POLYMERASE-CHAIN-REACTION ; DNA AMPLIFICATION ; SURFACE PASSIVATION ; BETA-LACTAMASE ; SYSTEM ; MICROARRAY ; SILICON ; NUMBER ; LOCUS
英文摘要:

Multiplex PCR is an effective tool for simultaneous multiple target detection but is limited by the intrinsic interference and competition among primer pairs when it is performed in one reaction tube. Dividing a multiplex PCR into many single PCRs is a simple strategy to overcome this issue. Here, we constructed a plastic, easy-to-use, fully sealed multiplex PCR chip based on reversible centrifugation for the simultaneous detection of 63 target DNA sequences. The structure of the chip is quite simple, which contains sine-shaped infusing channels and a number of reaction chambers connecting to one side of these channels. Primer pairs for multiplex PCR were sequentially preloaded in the different reaction chambers, and the chip was enclosed with PCR-compatible adhesive tape. For usage, the PCR master mix containing a DNA template is pipetted into the infusing channels and centrifuged into the reaction chambers, leaving the infusing channels filled with air to avoid cross-contamination of the different chambers. Then, the chip is sealed and placed on a flat thermal cycler for PCR. Finally, amplification products can be detected in situ using a fluorescence scanner or recovered by reverse centrifugation for further analyses. Therefore, our chip possesses two functions: 1) it can be used for multi-target detection based on end-point in situ fluorescence detection; and 2) it can work as a sample preparation unit for analyses that need multiplex PCR such as hybridization and target sequencing. The performance of this chip was carefully examined and further illustrated in the identification of 8 pathogenic bacterial genomic DNA samples and 13 drug-resistance genes. Due to simplicity of its structure and operation, accuracy and generality, high-throughput capacity, and versatile functions (i.e., for in situ detection and sample preparation), our multiplex PCR chip has great potential in clinical diagnostics and nucleic acid-based point-of-care testing.

语种: 英语
所属项目编号: 2012AA020101 ; 2012 M510464 ; 81327005 ; 81471749 ; 2012EP001002
项目资助者: National Hi-Tech Program of China ; China Postdoctoral Science Foundation ; National Natural Science Foundation of China ; National Sci-Tech Support Plan ; Beijing lab foundation
WOS记录号: WOS:000356372400013
Citation statistics:
内容类型: 期刊论文
URI标识: http://ir.bjmu.edu.cn/handle/400002259/50357
Appears in Collections:北京大学第二临床医学院_期刊论文

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作者单位: 1.Tsinghua Univ, Sch Med, Beijing 100084, Peoples R China
2.Collaborat Innovat Ctr Diag & Treatment Infect Di, Hangzhou 310003, Zhejiang, Peoples R China
3.Natl Engn Res Ctr Beijing Biochip Technol, Beijing 102206, Peoples R China
4.Peking Univ, Peoples Hosp, Beijing 100044, Peoples R China
5.CapitalBio Technol, Beijing 101111, Peoples R China

Recommended Citation:
Xu, Youchun,Yan, He,Zhang, Yan,et al. A fully sealed plastic chip for multiplex PCR and its application in bacteria identification[J]. LAB ON A CHIP,2015,15(13):2826-2834.
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