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1 6 2 Vol. 6 No Journal of Food Safety and Quality Feb., 2015 陈小金, 王乃福, 黄 * 晨, 董志珍 (, ) 摘要 :, (gene expression profiler genetic analysis system, GeXP),,, GeXP, GeXP, 关键词 : ; PCR; ; Development of gene expression profiler genetic analysis system and its application CHEN Xiao-Jin, WANG Nai-FU, HUANG Chen, DONG Zhi-Zhen * (Animal, Plant and Foodstuffs Inspection Center, Tianjin Entry-Exit Inspection and Quarantine Bureau, Tianjin , China) ABSTRACT: Detection of nucleic acid is widely used in life science, microbiology and medical science. With the developments of molecular biology, new testing approaches are constantly emerging on the basis of classical technology. As the regeneration technology of PCR and chip, gene expression profiler genetic analysis system (GeXP) has a broad application prospect, with features of high specificity and sensitivity, rapid speed and easy operation. This system truly realized the detection of high throughput for samples. This article reviewed the basic principle of GeXP system and its applications in veterinarian and animal husbandry, detection of genetically modified (GM) food and microorganism and medical research, which could provide a reference for detection of foodstuffs and microbiology. KEY WORDS: gene expression profiler genetic analysis system; multiplex-pcr; detection of foodstuffs; detection of microbe 1 引言 30,, Mullis PCR, [1],,, [2] PCR,, 基金项目 : (2014IK250) Fund: Supported by the Scientific and Technological Project of the General Administration of Quality Supervision, Inspection and Quarantine of the People s Republic of China (2014IK250) * 通讯作者 :,, baobao152@126.com *Corresponding author: DONG Zhi-Zhen, Researcher, Animal, Plant and Foodstuffs Inspection Center, Tianjin Exit-Entry Inspection and Quarantine Bureau, No.158 Jingmen Road, Free Trade Zone, Tianjin Port, Tianjin , China. wangnf@tjciq.gov.cn

2 2, : 411 PCR,, DNA PCR(multiplex PCR) [3], DNA,, DNA, Chamberhan 1988 PCR,, PCR [4-6] PCR, PCR,,, PCR,, [7], PCR RT-PCR [8-12] PCR,, PCR [13-16],,, (enzyme linked immunosorbent assay, ELISA) PCR [17-20], ELISA PCR ELISA,, PCR [21,22],, PCR,,, (gene expression profiler genetic analysis system, GeXP) PCR,, 2 GeXP 技术简介 2.1 GeXP 技术原理 GeXP PCR,, RNA, PCR PCR,, RNA GeXP PCR, RNA, cdna ;, cdna PCR, ;,,, 1 [23] PCR, PCR (DNA size standard, DSS),, [24] 2.2 GeXP 技术优缺点 GeXP,, GeXP,, GeXP 0.5, (0.5, R 2 >0.99) GeXP 30, 5760, GeXP PCR, PCR GeXP 30, 192, GeXP DNA (single nucleotide polymorphism, SNP) (short tandem repeat, STR) (loss of heterozigosity, LOH) (multiplex ligation-dependent probe amplification, MLPA), 5~50 ng RNA,, GeXP GeXP, PCR,,,

3 412 6 Fig. 1 1 GeXP [23] The principle sketch map of GeXP 3 GeXP 技术的应用 GeXP, DNA, GeXP, 3.1 GeXP 技术在畜牧兽医研究中的应用,, ( ), [25] H5 H7 H9 9 GeXP,, 38, 100 copies/μl 9,, Qin [26] GeXP H1N1 H1N1 H3N2,, H1N1 10,, [27,28] PCR GeXP,,,, 3.2 GeXP 技术在转基因食品检测中的应用 [29], GeXP 6 (PAT NOS CaMV35S, FMV35S Bar EPSPS), PCR [30], 100 ng ( ), PCR 3.3 GeXP 技术在微生物检测中的应用 GeXP Nagel [31] RT-PCR GeXP,, 20, 5 RT-PCR 68 [32] PCR GeXP

4 2, : 413,, GeXP, GeXP,, [33] GeXP - (mrt-pcr), (Japanese encephalitis virus, JEV) 8, 10 2 copies/μl 8 ( 13 ) RNA, [34] GeXP RT-PCR 12, A B H1N1 1~3 A B [35] 9 GeXP, 71 (HEV71) A (CVA) B (CVB)1 3 5, 10 3 copies/μl 9 RNA [36] GenomeLab GeXP A GI GII II 7 [37] 11 HPV GeXP PCR, (HPV) 3.4 GeXP 技术在医学研究中的应用 [38] GeXP 6, rhil-24 Alex [39] GeXP 70, 35, GeXP [40] GeXP, β-actin,, GeXP, [41] β- (β-actin) 14(SPR14) 18S-rRNA 3 3 CAD β-actin, SPR14, 18S-rRNA, β-actin SPR14 CAD [42] BCL6 KLF5 NCL (acute lymphoblastic leukemia, ALL) GeXP BCL6 KLF5 NCL ALL [43] GeXP, 7, 4 结语, PCR,,,,,,,, [24] PCR GeXP (GeXP),, PCR GeXP, DNA PCR GeXP PCR,,,, GeXP,,, GeXP,,,, 参考文献 [1] Saiki RK, Scharf S, Faloona F, et al. Enzymatic amplification of beta-globin genomic sequences and restriction site analysis for diagnosis of sickle cell anemia [J]. Science, 1985, 230: [2] Brenner S, Lerner RA. Encoded combinatorial chemistry [J]. Proc Natl Acad Sci USA, 1992, 89: [3] Chamberlain JS, Gibbs RA, Ranier JE, et al. Deletion screening of the Duchenne muscular dystrophy locus via multiplex DNA amplification [J]. Nucleic Acids Res, 1988, 16: [4] Caudai C, Padula MG, Bettini V, et al. Detection of HCV and GBV-C/HGV infection by multiplex PCR in plasma samples of transfused subjects [J]. J Virol Methods, 1998, 70: [5] Heredia A, Soriano V, Weiss SH, et al. Development of a multiplex PCR assay for the simultaneous detection and discrimination of HIV-1, HIV-2,

5 414 6 HTLV-I and HTLV-II [J]. Clin Diagn Virol, 1996, 7: [6] Chamberlain JS, Gibbs RA, Ranier JE, et al. Deletion screening of the Duehenne muscular dystrophy locus via multiplex DNA amplification [J]. Nucleic Acids Res, 1988, 16(23): [7],,,. PCR [J]., 2011, 9(8): Hou RS, Wang L, Zhang Q, et al. Multiplex PCR technique and its application in pathogen detection [J]. Chin J Gen Pract, 2011, 9(8): [8] Flajshans M, Cossonc J, Redina M. The application of image cytometry to viability assessment in dual fluorescence-stained fish spermatozoa [J]. Cell Biol Int, 2004, 28(12): [9] Keer JT, Birch L. Molecular methods for the assessment of bacterial viability [J]. J Microbiol Meth, 2003, 53(2): [10] Vijay KS. Real-time reverse transcription-multiplex PCR for simultaneous and specific detection of rfbe and eae genes of Escherichia coli O157:H7 [J]. Mol Cell Probe, 2006, 20 (5): [11] Narjol GE, Axe F, Carlos AG, et al. 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Detection of pandemic influenza A H1N1 virus by multiplex reverse transcription-pcr with a GeXP analyzer [J]. J Virolo Meth, 2010, 168: [27]. [D]. :, Fang YB. Cloning and expression profile of two immune-related genes and studies on the applicability of eleven internal reference genes in Mud Crab, Scylla paramamosain [D]. Shanghai: Shanghai Ocean University, [28]. [D]. :, Sun MM. Cloning and expression profile of the response to stress genes from mud crab, Scylla Paramamosain and Eriocheir sinensis [D]. Shanghai: Shanghai Ocean University, [29]. GeXP-PCR HPV [D]. :, Yang MJ. Detection of genetically modified foods and genotyping HPV infection by a GeXP based Multiplex PCR Assay [D]. Guangzhou: Jinan University, [30] SN/T , PCR [S]. :, SN/T , Protocol of the real-time PCR for detecting genetically modified plants and their derived products [S]. Beijing: Standards Press of China, [31] Nagel M, Gilden D, Shade T, et al. 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6 2, : 415 [J]., 2012, 28(1): He B, Wang HY, Zhang C, et al. Development of a GeXP based multiplex RT-PCR assay for simultaneous detection of eight arboviruses related to encephalitis [J]. Chin J Virol, 2012, 28(1): [34],,,. GeXP PCR 12 [J]., 2011, 27(6): Li J, Mao NY, Qin M, et al. A GeXP Based Multiplex RT-PCR Assay for Simultaneous Detection of twelve human respiratory virus [J]. Chin J Virol, 2011, 27(06): [35],,,. GeXP [J]., 2011, 27(4): Hu XM, Zhang Y, Xu BL, et al. Development of a GeXP based multiplex RT-PCR assay for simultaneous differentiation of nine human hand food mouth disease pathogens [J]. Chin J Virol, 2011, 27(04): [36],,,. [J]., 2011, 27(3): Liu Y, Xu ZQ, Li JS, et al. A Novel method for multiplex detection of gastroenteritis-associated viruses [J]. Chin J Virol, 2011, 27(03): [37],,,. GeXP., 2013, 47(12): Yang MJ, Luo L, Nie K, et al. Detection and typing of human papillomavirus by a GeXP Assay [J]. Chin J Prev Med, 2013, 47(12): [38],,,. rhil-24 [J]., 2013, 29(12): Wang SH, Guo JJ, Tang YL, et al. The detection of apoptosis-associated genes in SKOV3, SKOV3 /ddp cells of the recombinant human interleukin-24 induced by GeXP analysis system [J]. Chin J Immunol, 2013, 29(12): [39] Alex JR, Rashmi MK, William G, et al. Analytical validation of the GeXP analyzer and design of a workflow for cancer-biomarker discovery using multiplexed gene-expression profiling [J]. Anal Bioanal Chem, 2009, 393: [40],,,. GeXP [J]., 2009, 16 (1): Dong J, Tian YP, Zhang YD, et al. Optimized detection of peripheral blood inflammatory related gene expression by GeXP multi gene expression system [J]. Label Immunoass Clin Med, 2009, 16 (1): [41],,,. [J]., 2010, 6 (17): Zhang YD, Wen XY, Dong J, et al. Comparison of housekeeping genes expression levels in analysis of coronary artery disease related genes [J]. Label Immunoass Clin Med, 2010, 6 (17): [42],,,. BCL6 KLF5 NCL [J]., 2011, 79(4): Zhu L, Zheng HY, Liu X, et al. Aberrant expression of BCL6, KLF5 and NCL in pediatric patients with acute lymphoblastic leukemia [J]. Chin J Cancer Biother. 2011, 79(04): [43]. GeXP [J]., 2014, 91(7): Li D. Using a GeXP system to detect genes responsible for the multidrug resistance of klebsiella pneumonia in patients with sepsis [J]. J Pathog Biol, 2014, 91(7): 作者简介 ( 责任编辑 : 张宏梁 ) 陈小金, 兽医师, 主要研究方向为动物传染病诊断与防治新技术 chenxj0@tjciq.gov.cn 董志珍, 研究员, 主要研究方向为动物疫病检测技术 baobao152@126.com

Microsoft Word 聂雪梅.doc

Microsoft Word 聂雪梅.doc 4 4 Vol. 4 No. 4 2013 8 Journal of Food Safety and Quality Aug., 2013 聂雪梅 *, 李立, 孙利, 高飞 (, 100123) 摘要 : 2011 12,, 2007~2011,,,, 关键词 : ; ; ; ; Trade and notification situation on food import and export