1、| | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | BRITISH STANDARD BS EN 12305:1998 The Euro
2、pean Standard EN 12305:1997 has the status of a British Standard ICS 07.080 NO COPYING WITHOUT BSI PERMISSION EXCEPT AS PERMITTED BY COPYRIGHT LAW Biotechnology Modified organisms for application in the environment Guidance for the sampling strategies for deliberate releases of genetically modified
3、plantsThis British Standard, having been prepared under the direction of the Sector Board for Materials and Chemicals, was published under the authority of the Standards Board and comes into effect on 15 March 1998 BSI 1998 ISBN 0 580 29263 0 BS EN 12305:1998 Amendments issued since publication Amd.
4、 No. Date Text affected National foreword This British Standard is the English language version of EN 12305:1997. The UK participation in its preparation was entrusted to Technical Committee CII/58, Biotechnology, which has the responsibility to: aid enquirers to understand the text; present to the
5、responsible international/European committee any enquiries on the interpretation, or proposals for change, and keep the UK interests informed; monitor related international and European developments and promulgate them in the UK. A list of organizations represented on this committee can be obtained
6、on request to its secretary. Cross-references The British Standards which implement international or European publications referred to in this document may be found in the BSI Standards Catalogue under the section entitled “International Standards Correspondence Index”, or by using the “Find” facili
7、ty of the BSI Standards Electronic Catalogue. A British Standard does not purport to include all the necessary provisions of a contract. Users of British Standards are responsible for their correct application. Compliance with a British Standard does not of itself confer immunity from legal obligati
8、ons. Summary of pages This document comprises a front cover, an inside front cover, the EN title page, pages 2 to 7 and a back cover.CEN European Committee for Standardization Comite Europe en de Normalisation Europa isches Komitee fu r Normung Central Secretariat: rue de Stassart 36, B-1050 Brussel
9、s 1997 CEN All rights of exploitation in any form and by any means reserved worldwide for CEN national Members. Ref. No. EN 12305:1997 E EUROPEAN STANDARD EN 12305 NORME EUROPE ENNE EUROPA ISCHE NORM October 1997 ICS 07.080 Descriptors: Biotechnology, genetics, plants, botany, modified organisms, de
10、finitions, sampling, environmental protection English version Biotechnology Modified organisms for application in the environment Guidance for the sampling strategies for deliberate releases of genetically modified plants Biotechnologie Organismes modifie s disse mine s dans lenvironnement Guide des
11、 strate gies de chantillonnage pour les disse minations volontaires de plantes ge ne tiquement modifie es. Biotechnik Vera nderte Organismen zum Einsatz in der Umwelt Leitfaden fu r Probenahmestrategien bei der absichtlichen Freisetzung gentechnisch vera nderter Pflanzen This European Standard was a
12、pproved by CEN on 21 August 1997. CEN members are bound to comply with the CEN/CENELEC Internal Regulations which stipulate the conditions for giving this European Standard the status of a national standard without any alteration. Up-to-date lists and bibliographical references concerning such natio
13、nal standards may be obtained on application to the Central Secretariat or to any CEN member. This European Standard exists in three official versions (English, French, German). A version in any other language made by translation under the responsibility of a CEN member into its own language and not
14、ified to the Central Secretariat has the same status as the official versions. CEN members are the national standards bodies of Austria, Belgium, Czech Republic, Denmark, Finland, France, Germany, Greece, Iceland, Ireland, Italy, Luxembourg, Netherlands, Norway, Portugal, Spain, Sweden, Switzerland
15、and United Kingdom.Page 2 EN 12305:1997 BSI 1998 Foreword This European Standard has been prepared by Technical Committee CEN/TC 233, Biotechnology, the secretariat of which is held by AFNOR. This European Standard shall be given the status of a national standard, either by publication of an identic
16、al text or by endorsement, at the latest by April 1998, and conflicting national standards shall be withdrawn at the latest by April 1998. This European Standard has been prepared under a mandate given to CEN by the European Commission and the European Free Trade Association, and supports essential
17、requirements of EU Directive(s). According to the CEN/CENELEC Internal Regulations, the national standards organizations of the following countries are bound to implement this European Standard: Austria, Belgium, Czech Republic, Denmark, Finland, France, Germany, Greece, Iceland, Ireland, Italy, Lux
18、embourg, Netherlands, Norway, Portugal, Spain, Sweden, Switzerland and the United Kingdom. Contents Page Foreword 2 1 Scope 3 2 Normative references 3 3 Definitions 3 4 General consideration 4 5 Sampling strategy 4 Annex A (informative) Relationship between a valid sampling and monitoring strategy 6
19、 Annex B (informative) Bibliography 7Page 3 EN 12305:1997 BSI 1998 Introduction When the genetically modified plants are subject to experimental release into the environment, it is important to ensure the validity of sampling strategies used to monitor the release. The sampling strategy and the stat
20、istical analysis used in the deliberate release of genetically modified plants depend on the frequency of occurrence and spatial distribution of the relevant genetically modified plants and/or introduced gene in the experimental field studied. Since there are many different techniques available for
21、the detection and the identification of genetically modified plants, this European Standard is intended to give guidance to the experimenter on the design of a sampling strategy appropriate to the field trial, to the plants and to the particular traits being used. Not all of the points are necessari
22、ly relevant to a particular experiment. The principles can also be applied to sampling of plants grown in greenhouses. This European standard gives the experimenter a list of points that should be considered in determining the validity of a sampling strategy comprising valid design, review, executio
23、n and documentation of a sampling protocol. 1 Scope This European Standard gives guidance for setting up a valid sampling strategy to meet the objectives of a monitoring strategy for genetically modified plants. The sampling is to provide material to which subsequent analytical methods for monitorin
24、g genetically modified plants of interest can be applied (see prEN 12468). This European Standard deals only with providing statistically valid samples from the experimental field as identified in a field trial design. The mode of sampling is dependent on purpose. Sampling may be carried out to meet
25、 the objectives of the investigator and/or statutory requirements. Therefore this European Standard provides the experimenter with a list of guiding parameters that should be considered in determining the validity of the proposed strategy for sampling. 2 Normative references This European Standard i
26、ncorporates by dated or undated reference, provisions from other publications. These normative references are cited at the appropriate places in the text and the publications are listed hereafter. For dated references, subsequent amendments to or revisions of any of these publications apply to this
27、European Standard only when incorporated in it by amendment or revision. For undated references the latest edition of the publication referred to applies. prEN 12468, Biotechnology Modified organisms for application in the environment Guidance for the monitoring strategies for deliberate releases of
28、 genetically modified plants 3 Definitions For the purposes of this standard, the following definitions apply. 3.1 behaviour interaction of the organism(s) with abiotic and biotic environments, its (their) occurrence, persistence and spreading abilities 3.2 control sample or preparation of known cha
29、racteristics used to standardize the analysis 3.3 experimental field area within a release site which contains the plots necessary to standardize the analysis 3.4 field trial design specifications for the planting of a trial, i.e. number and layout of plots in the field NOTE The field trial design i
30、nfluences the type of statistical analysis that can be performed and vice versa. 3.5 genetically modified plant plant in which the genetic material has been altered in a way that does not occur naturally by mating and/or natural recombination NOTE Within the terms of this definition, genetic modific
31、ation occurs at least through the use of the techniques listed in the Directive 90/220/EEC or its appropriate annexes, see annex B 1. 3.6 monitoring regular or continuous supervision and checking of an organism, process or procedure NOTE In this standard, monitoring applies to the progress of a rele
32、ased genetically modified plant. 3.7 monitoring strategy procedure for designing, reviewing, executing and documenting a monitoring protocol 3.8 plot area where an individual experimental unit or treatment is carried out 3.9 release site defined area which contains one or more experimental field NOT
33、E Several different trials can occur within a release site. 3.10 sample materials collected for analysisPage 4 EN 12305:1997 BSI 1998 3.11 sampling protocol written document describing the manner in which samples are collected, transported, stored, and their pretreatment 3.12 sampling strategy proce
34、dure for designing, reviewing, executing and documenting a sampling protocol 3.13 trait observable and/or measurable characteristic 4 General considerations To ensure that the design of a sampling strategy is valid, the samples collected from a release should be representative of each experimental f
35、ield. The number of samples required for representative sampling depends on the field trial design and experimental aims. The field trial design also determines the types of statistical analysis that can be performed. The following types of field trials can be used. a) Trials to study the impact of
36、an introduced trait in a genetically modified crop or to obtain lines containing the introduced trait. Typically such trials require a relatively small number of samples, since the plants tested are supposed to be similar. b) Trials to study the transfer of a gene between plants. Typically such tria
37、ls require a larger number of samples for analysis to detect rare events. c) Trials to investigate the behaviour of an introduced trait in the genetically modified plants. In these trials, the sampling strategy depends on the trait studied. The stability of the trait is an important consideration. N
38、OTE 1 Guidance for points to consider for the analysis of the stability of an introduced trait is given in prEN 12683, see annex B 2. The presence of adequate control samples is essential for the validation of results. The design of most field trials with genetically modified plants will follow the
39、same principles as applied to field trials with plants which are not genetically modified. Statistical analysis required for quantitative evaluation influences the sampling strategy and hence the validity of the results. The purpose of the experiment should be defined in order to prepare an adequate
40、 protocol for sampling. Specific requirements of any subsequent detection and identification methods should be considered and will influence the utility of the results for monitoring purposes (see prEN 12468). The sampling strategy should be re-evaluated periodically to ensure its validity. NOTE 2 T
41、he development of a valid monitoring and sampling strategy for deliberate releases of genetically modified plants in the environment is summarized in Figure A.1. 5 Sampling strategy 5.1 General It is first necessary to determine the objectives of the sampling strategy. The main steps in the developm
42、ent of the sampling strategy are: a) statement of experimental objectives; b) design and review of sampling protocol; c) execution of the sampling protocol d) appropriate record keeping. Responsibilities should be assigned for each step to a specific person. The sampling strategy should be reviewed
43、regularly, in the light of field inspections, to ensure its continuing validity. 5.2 Design criteria The design of the sampling protocol should meet defined objectives as determined by the investigator and/or by statutory requirements. The design can vary widely, for example, requirements for trials
44、 of introduced traits see 4a), for studies of gene transfer see 4b) or gene behaviour see 4c) can differ. The protocol for sampling should be part of the preparation and design of the experiment, keeping the flexibility that could be required by unexpected observations, and should therefore be suppo
45、rted by a clear design of the experiment. The sampling protocol should be documented. Parameters such as when sampling will take place, the location(s) and/or site size, the number of genotypes, number of plots and means of plot identification and the number of treatment(s) are important in the deve
46、lopment of the design. There are many different techniques available for the detection and identification of genetically modified plants. The following should be considered in the initial phase of the design: will samples be used for qualitative or quantitative analysis? will sampling be for express
47、ed characteristics and/or target nucleic acids? which identification and/or detection methods will be applied to the samples? is the sampling destructive? is the protocol practical and feasible? The expression of characteristics in genetically modified plants is further dependent on the following fa
48、ctors: genetic background; developmental stage of the plant; plant part of relevance such as whole plant, root, leaf, flower, stem, pollen, seed; specific environmental or external constraints on the behaviour of the traits.Page 5 EN 12305:1997 BSI 1998 Therefore, the following should be considered
49、in the design of a representative and statistically valid sampling protocol: choice of appropriate statistical method; required level of significance; timing and frequency of sampling; part of the plant to be sampled; separate or pooled samples; stability of sample; number and size of samples; number of plots; number of plants; number of samples per plot; number of samples per plant; appropriate negative and/or positive controls. The labelling of the samples and the sam
