7 2 Vol. 7 No. 2 2016 2 Journal of Food Safety and Quality Feb., 2016 蔡 1, 军 2# 1,, 李慧 2#* 1,, 胡梦龙 2 1,, 傅洋 2, 汪磊 3, 王璐 4 (1., 102209; 2., 102209; 3. ( ), 433000; 4., 100020) 摘要 :,,,,,,, PCR PCR,,, 关键词 : ; ; Research progress in techniques for the detection and analysis of genetically modified ingredients CAI Jun 1, 2#, LI Hui 1, 2#*, HU Meng-Long 1, 2, FU Yang 1, 2, WANG Lei 3, WANG Lu 4 (1. Nutrition & Health Research Institute, COFCO Corporation, Beijing 102209, China; 2. Beijing Key Laboratory of Nutrition, Health & Food Safety, Beijing 102209, China; 3. Quality & Safety Department, COFCO Rice (Xiantao) Co. Ltd., Xiantao 433000, China; 4. Quality & Safety Management Department, COFCO Rice Division, Beijing 102209, China) ABSTRACT: With the rapid application and development of transgenic technology in agriculture, the categories and quantities of transgenic products are progressively increased, the safety of genetically modified organisms (GMOs) has attracted more and more extensive concern of the public, the genetically modified productions labeling systems all over the world has been gradually established and improved, and some countries have provided a minimum threshold of genetically modified ingredients (GMIs) content. Accordingly, it is very urgently needed constant innovation and development for the transgenic detection technique systems. The article briefly reviewed the common detection technology and method of GMIs both in China and abroad, the existing primary detection technology and method of GMIs mainly based on detection of exogenous protein and nucleic acid, such as ELISA, PCR, isothermal amplification technology, gene chip technology, digital PCR and so on. It also summarized the application progress of the corresponding in the detection of GMIs, meanwhile summarized the advantages and disadvantages of each corresponding methods, in order to be aware clearly and completely in public about the current situation of global # # CAI Jun and LI Hui are co-first authors. * 通讯作者 :,,, E-mail: lhui@cofco.com *Corresponding author: LI Hui, Ph.D., Senior Engineer, Nutrition & Health Research Institute, COFCO Corporation, Beijing 102209, China. E-mail: lhui@cofco.com
2, : 707 detection technologies of GMO and their future development trend. KEY WORDS: genetically modified organisms; common detection technique; research progress 1 引言 1994,,,, 2014 28, 1996 170 2014 1.81, 100 [1],, [2] [3] [4] MON863 [5] Mon810 [6] CryAb1 (Bt ) [7],,,,,,,, [8-10],, (enzyme-linked immunosorbent assay, ELISA) PCR (isothermal amplification) (gene chip) PCR (digital PCR, d PCR), 2 常见转基因检测技术 2.1 基于外源蛋白靶标的检测技术, (ELISA) (Western blot) (protein chip) 2.1.1 酶联免疫吸附技术, Kim [11] PAT NPTⅡ ; Kamle [12] Bt ; Tan [13] CpT1, 0.21 ng/ml 2.1.2 Western blot 检测技术 Western blot SDS-PAGE,,,,, [14] Western blot EPSPS [15] Western Blot, CP4-EPSPS, 0.5%, Western blot (TSM) EPSPS Wang [16] Western blot EPSPS 2.1.3 免疫层析试纸条技术, [17] CP4 EPSPS,, [18] Bt Bt-Cry1Ab/Ac, Cry1Ab/Cry1Ac [19] 2.1.4 蛋白质芯片技术,, [20] 3 BT Cry1Ac
708 7 BT Cry1Ah, [21] 2.2 基于外源核酸成分的检测技术, DNA, DNA,,,,, DNA DNA,, [22] PCR PCR PCR 2.2.1 定性 PCR 检测技术 PCR PCR DNA, PCR PCR PCR PCR(nested and semi-nested PCR) PCR (multiplex PCR) PCR (multiplex real-time PCR) (1) PCR PCR,, [23,24], DNA PCR, DNA, PCR, (2) PCR PCR PCR PCR,,, SYBR Green SYTO9 LC Green Evagreen, Taqman Dual Probes, Taqman Taqman PCR PCR, PCR, [25] Dinon [26] cry1a.105 cry2ab2 PCR, 0.01 0.05 ng; Moor [27] FMV DNA PCR, FMV 34S ; Grohmann [28] bar ctp2-cp4epsps PCR, 0.02% PCR,, (3) PCR DNA, PCR PCR PCR PCR,,,, PCR PCR,,, [29] PCR, 0.001% 0.01%, [30] PCR, [31] Ca MV35S Nos PCR, 0.01% 0.05% (4) PCR PCR, PCR, PCR 2, PCR, PCR,, Yoke-Kqueen [32] EPSPS Cry1Ab PCR, [33] 7 PCR, [34] PCR, 0.2 ng, [35] 3 PCR, 0.9%(V/V) (5) PCR PCR
2, : 709, PCR, PCR PCR PCR PCR, Bahrdt [36] 6 PCR, 1000 ; Hans-Henno [37] CaMV35S NOS FMV35S bar PCR, 10 ; Cottenet [38] 47 24 PCR, 1~16 ; Chaouachi [39] 4 (Bt11, Bt176, Mon810 T25) PCR ; Köppel [40] PCR, 0.1% 2.2.2 定量 PCR 检测技术,, 0.9%( ) 0.5%( ) [41], 1% [42], ( 5 ) 3% [43], ( 3 ) 5% [44], 5% [45], PCR,,, PCR 3 : PCR PCR (quantitative competitive PCR, QC-PCR) PCR (real-time fluorescence quantitative PCR) (1) PCR PCR PCR-ELISA,, PCR,, PCR-ELISA PCR-ELISA PCR ELISA,, [46] PCR-ELISA, [47] PCR-ELISA, [48] PCR-ELISA PCR-ELISA PCR,, DNA, (2) PCR PCR PCR, DNA, DNA DNA, DNA, DNA Eri [49] PCR Anastasia [50] PCR PCR, - DNA [51] (3) PCR PCR PCR PCR,, PCR, [52] 59122 PCR, [53] GHB119 TaqMan PCR, 10, [54] PCR PCR,, PCR, 2.2.3 等温扩增技术 PCR, (LAMP) (cross priming amplification, CPA) (strand displacement amplification, SDA) (rolling circle amplification, RCA) DNA ( helicase-dependent isothermal DNA amplification, HDA) (nucleic acid sequence-based amplification, NASBA) (rapid isothermaldetection and amplification, RIDA) (nicking endonuclease mediated amplification, NEMA) (transcription mediated amplification) (isothermal multiple self-matching-initiated amplification, IMSA) (LAMP) (CPA)
710 7 (1) (LAMP) 2000 Notomi [55], 6 4, Bst DNA,,, PCR,, [56],, [57] BT 11, 0.5%; [58] CaMV35S, 200 /μl; [59] LY038, 0.01 ; [60] A2704-12, 0.1%; [61] J163, 16 pg,, (2) (CPA),, Bst DNA,,, PCR,, [62] Bt63, 0.1%; [63] CaMV35S, 0.05%; [64] NOS, 0.05%, (3), [65] ; [66] 6 RPA, 500 ; [67] Cry1Ab/c RPA, 0.1% 2.2.4 基因芯片 (gene chip) 技术 DNA DNA (DNA microarray), 20 80 DNA DNA,,,, DNA,, Turkec [68] 12, 10 1%; Seong-Hun [69], 0.5%; [70,71], 0.1%,,, 2.2.5 数字 PCR(digital PCR, dpcr) 技术 PCR PCR,, PCR PCR,, ; PCR,,, Fluidigm Life Technologies Bio-Rad RainDance PCR, microrna NGS [72] PCR TT51-1 ; Dany [73] PCR ; [74] PCR
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