KS J ISO 21572-2009 Foodstuffs－Methods for the detection of genetically modified organisms and derived products－Protein based methods《食品 基因改良有机物及衍生产品的检测方法 基于蛋白质的方法》.pdf

1、 KSKSKSKS KSKSKSK KSKSKS KSKSK KSKS KSK KS KS J ISO 21572 KS J ISO 21572 :2009 2009 12 10 http:/www.kats.go.krKS J ISO 21572:2009 : ( ) ( ) () () ( ) : (http:/www.standard.go.kr) ： ： 2004 11 30 ： 2009 12 10 2009-0785 ： ： ( 02-509-7266) (http:/www.kats.go.kr). 10 5 , . KS J ISO 21572:2009 i .1 1 1 2

2、1 3 .1 4 6 5 6 6 .6 7 6 8 6 8.1 .6 8.2 .6 8.3 .7 8.4 (calibration curve) 7 8.5 7 9 7 9.1 .7 9.2 8 9.3 8 10 .8 10.1 8 10.2 .8 11 .9 12 .10 A() (Roundup Ready 1) 11 22 KS J ISO 21572:2009 FoodstuffsMethods for the detection of genetically modified organisms and derived productsProtein based methods 20

3、04 1(2005 ) ISO 21572, FoodstuffsMethods for the detection of genetically modified organisms and derived productsProtein based methods , . 1 / . . A . . 2 . . ( ) . prEN ISO 21568, FoodstuffsMethods of analysis for the detection of genetically modified organisms and derived productsSampling 3 . 3.1

4、3.1.1 (sample) KS J ISO 21572:2009 2 KS Q ISO 35341：2009 3.1.2 (laboratory sample) 3.1.3 ( ) test sample(test portion) KS Q ISO 35341 3.1.4 ( , matrix) . . 3.1.5 (denaturation of proteins) / ( ) . , , . 3.2 3.2.1 (antibody) B ( ). . 3.2.2 (antigen) 3.2.3 (clone) 3.2.4 (cross-reactivity) . 3.2.5 (mon

5、oclonal antibody) (hybridoma) (antigen determinant) 3.2.6 (polyclonal antibody) KS J ISO 21572:2009 3 3.2.7 (specificity of an antibody) 3.3 3.3.1 (conjugate) (： ), . 3.3.2 (Western blotting) (： ) . . 3.3.3 ELISA ( ：enzyme linked immunosorbent assay) . . 3.3.4 (test kit) , 3.3.5 (dip stick format) l

6、ateral flow 3.4 3.4.1 (reference material) , KS A ISO Guide 30 3.4.2 (standard) , , , , 3.5 KS J ISO 21572:2009 4 3.5.1 (accuracy) (bias) . KS Q ISO 35341：2009 3.5.2 (precision) 1 . 2 . . 3 . . . KS Q ISO 35341：2009. 3.5.3 (bias) () 3.5.4 (sensitivity) . . 3.5.5 (specificity) Codex 3.5.6 (limit of d

7、etection：LOD) 1 . 1, 2 3.5.7 (limit of quantitation：LOQ) KS A ISO 5725, 2 3 1 . 3.5.8 ( / ) applicability range(range of quantification/linearity/ dynamic range) KS J ISO 21572:2009 5 ( ) 3.5.9 (repeatabilityreproducibility limit) 95 % rR KS Q ISO 35341 , rR2.8 s r s R . 3.5.10 (reproducibility) KS

8、Q ISO 35341：2009 3.5.11 (reproducibility conditions) KS Q ISO 35341：2009 3.5.12 (repeatability) KS Q ISO 35341：2009 3.5.13 (repeatability conditions) KS Q ISO 35341：2009 3.5.14 (repeatabilityreproducibility standard deviation) . “ (variance)” “ ” . KS J ISO 21572:2009 6 3.5.15 (recovery) (fortified)

9、 4 , . 5 , , . , , , . , , , . 6 A.5 . 7 prEN ISO 21568 . 8 8.1 , , . , , (blanks), , (calibration curve) . . ( ) . 8.2 A . . / . KS J ISO 21572:2009 7 . , A . ( ) . . 8.3 . /, A . , ( ). 8.4 (calibration curve) . 8.5 A , , / (duplicate) . . , . (stability) . . 9 9.1 , . , 15 % . 20 % . % , . 3 (wel

10、l) . GMO . “ ” “ ” . “ ” . KS J ISO 21572:2009 8 9.2 , , , % ( ) . . . 9.3 . . 10 10.1 A , . . . ( % ) , . . . . 10.2 10.2.1 () . . non-GMO non-GMO GMO . . A . 10.2.2 KS J ISO 21572:2009 9 . . . . , (calibration) . 10.2.3 . . . , () . . . . . . 10.2.4 / , . (calibration points) ( ). “ ” , . 10.2.5 .

11、 , . 10.2.6 (calibrants) . . . 11 ( ) , HPLC . / . . KS J ISO 21572:2009 10 12 . , , ( ) KS J ISO 21572:2009 11 A () (Roundup Ready 1) ) A.1 (Glycine max) 2) . cp4 epsps . (glyphosate) Agrobacterium sp. strain CP4 . CP4 EPSPS Roundup 1) . CP4 EPSPS . A.2 Roundup Ready 1) GTS 4032 CP4 EPSPS ELISA . C

12、P4 EPSPS . , . , . (IRMM ) CP4 EPSPS (0.52) %(w/w ) 3) . 4) . 3, 4 . A.3 CP4 EPSPS A.1A.4 ELISA . 1) Roundup Ready Roundup (Monsanto Company) . CEN(European Committee for Standardzation) . 2) , , , . 3) European Commissions Joint Research Center, Institute for Reference Materials and Measurements(IR

13、MM) . . 4) Strategic Diagnostics, Inc. Hampshire, England. . KS J ISO 21572:2009 12 1 2 A.1 . 1 2 A.2 . . 1 HRP 2 3 A.3 CP4 EPSPS HRP(horseradish peroxidase) . 1 A.4 HRP(horseradish peroxidase) TMB(tetramethylbenzidine) . HRP . . 450 nm . KS J ISO 21572:2009 13 A.4 A.4.1 . . . . (28) . . (28) 9 . (A

14、.6.6.1) (A.6.6.2) (28) . (28) . A.4.2 A.4.2.1 , pH 7.5 A.4.2.2 PBS, Tween 20, bovine serum albumin, pH 7.4 A.4.2.3 (well) 8 (well) 12 A.4.2.4 horseradish peroxidase(HRP) rabbit anti-CP4 EPSPS A.4.2.5 10 % A.4.2.6 K-Blue TM5) tetramethylbenzidine(TMB), , 5 % dimethylformamide A.4.2.7 0.5 % A.4.2.8 10 PBS, Tween 20, pH 7.1 A.4.2.9 0.1 %, 0.5 %, 1 %, 2 % 5 % w/w A.4.3 A.4.3.1 70 % 95 % A.4.3.2 A.5 5) K-Blue Neogen Corp, Lansing, Michigan, USA . CEN . , . KS J ISO 21572:2009 14 A.5.1 A.5.2 4 A.5.3 (conical) , 15 mL A.5.4 A.5.5