BS PD CEN TS 16826-1-2015 Molecular in vitro diagnostic examinations Specifications for pre-examination processes for snap frozen tissue Isolated RNA《分子体外诊断检查 快速冷冻组织的预检测过程.pdf

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1、BSI Standards Publication PD CEN/TS 16826-1:2015 Molecular in vitro diagnostic examinations Specifications for pre-examination processes for snap frozen tissue Part 1: Isolated RNAPD CEN/TS 16826-1:2015 PUBLISHED DOCUMENT National foreword This Published Document is the UK implementation of CEN/TS 1

2、6826-1:2015. The UK participation in its preparation was entrusted to Technical Committee CH/212, IVDs. A list of organizations represented on this committee can be obtained on request to its secretary. This publication does not purport to include all the necessary provisions of a contract. Users ar

3、e responsible for its correct application. The British Standards Institution 2015. Published by BSI Standards Limited 2015 ISBN 978 0 580 85026 4 ICS 11.100.10 Compliance with a British Standard cannot confer immunity from legal obligations. This Published Document was published under the authority

4、of the Standards Policy and Strategy Committee on 31 August 2015. Amendments issued since publication Date Text affectedPD CEN/TS 16826-1:2015TECHNICAL SPECIFICATION SPCIFICATION TECHNIQUE TECHNISCHE SPEZIFIKATION CEN/TS 16826-1 August 2015 ICS 11.100.10 English Version Molecular in vitro diagnostic

5、 examinations - Specifications for pre-examination processes for snap frozen tissue - Part 1: Isolated RNA Tests de diagnostic molculaire in vitro - Spcifications relatives aux processus pranalytiques pour les tissus conglation rapide - Partie 1: ARN extrait Molekularanalytische in-vitro-diagnostisc

6、he Verfahren - Spezifikationen fr pranalytische Prozesse fr schockgefrorene Gewebeproben - Teil 1: Isolierte RNS This Technical Specification (CEN/TS) was approved by CEN on 6 July 2015 for provisional application. The period of validity of this CEN/TS is limited initially to three years. After two

7、years the members of CEN will be requested to submit their comments, particularly on the question whether the CEN/TS can be converted into a European Standard. CEN members are required to announce the existence of this CEN/TS in the same way as for an EN and to make the CEN/TS available promptly at

8、national level in an appropriate form. It is permissible to keep conflicting national standards in force (in parallel to the CEN/TS) until the final decision about the possible conversion of the CEN/TS into an EN is reached. CEN members are the national standards bodies of Austria, Belgium, Bulgaria

9、, Croatia, Cyprus, Czech Republic, Denmark, Estonia, Finland, Former Yugoslav Republic of Macedonia, France, Germany, Greece, Hungary, Iceland, Ireland, Italy, Latvia, Lithuania, Luxembourg, Malta, Netherlands, Norway, Poland, Portugal, Romania, Slovakia, Slovenia, Spain, Sweden, Switzerland, Turkey

10、 and United Kingdom. EUROPEAN COMMITTEE FOR STANDARDIZATION COMIT EUROPEN DE NORMALISATION EUROPISCHES KOMITEE FR NORMUNG CEN-CENELEC Management Centre: Avenue Marnix 17, B-1000 Brussels 2015 CEN All rights of exploitation in any form and by any means reserved worldwide for CEN national Members. Ref

11、. No. CEN/TS 16826-1:2015 EPD CEN/TS 16826-1:2015 CEN/TS 16826-1:2015 (E) 2 Contents Page European foreword .3 Introduction .4 1 Scope 5 2 Normative references 5 3 Terms and definitions .5 4 General considerations .6 5 Outside the laboratory 7 5.1 Primary tissue collection manual.7 5.1.1 Information

12、 about the primary sample donor .7 5.1.2 Information on the primary tissue sample 7 5.1.3 Information on the primary tissue sample processing 8 5.2 Transport requirements 8 6 Inside the laboratory .9 6.1 Information on the primary tissue sample receipt .9 6.2 Evaluation of the pathology of the speci

13、men and selection of the sample.9 6.3 Cryo-storage of the specimen 9 6.4 Storage requirements . 10 6.5 Isolation of the total RNA . 11 6.5.1 General information for RNA isolation procedures 11 6.5.2 Using commercial kits 11 6.5.3 Using the laboratories own protocols . 12 6.6 Quality assessment of is

14、olated RNA 12 6.7 Storage of isolated RNA . 12 Annex A (informative) Impact of preanalytical variables on RNA profiles obtained from frozen liver tissue samples collected during and after routine surgery . 13 A.1 Comparison of stable and unstable genes identified under ischemic conditions 13 A.2 Rec

15、ommendations based on the results . 15 Bibliography . 16 PD CEN/TS 16826-1:2015 CEN/TS 16826-1:2015 (E) 3 European foreword This document (CEN/TS 16826-1:2015) has been prepared by Technical Committee CEN/TC 140 “In vitro diagnostic medical devices”, the secretariat of which is held by DIN. Attentio

16、n is drawn to the possibility that some of the elements of this document may be the subject of patent rights. CEN and/or CENELEC shall not be held responsible for identifying any or all such patent rights. According to the CEN-CENELEC Internal Regulations, the national standards organizations of the

17、 following countries are bound to announce this Technical Specification: Austria, Belgium, Bulgaria, Croatia, Cyprus, Czech Republic, Denmark, Estonia, Finland, Former Yugoslav Republic of Macedonia, France, Germany, Greece, Hungary, Iceland, Ireland, Italy, Latvia, Lithuania, Luxembourg, Malta, Net

18、herlands, Norway, Poland, Portugal, Romania, Slovakia, Slovenia, Spain, Sweden, Switzerland, Turkey and the United Kingdom. PD CEN/TS 16826-1:2015 CEN/TS 16826-1:2015 (E) 4 Introduction Molecular in vitro diagnostics has enabled a significant progress in medicine. Further progress is expected by new

19、 technologies analysing signatures of nucleic acids, proteins, and metabolites in human tissues and body fluids. However, the profiles and/or integrity of these molecules can change drastically during primary sample collection, transport, storage, and processing thus making the outcome from diagnost

20、ics or research unreliable or even impossible because the subsequent analytical assay will not determine the situation in the patient but an artificial profile generated during the pre-examination process. Therefore, a standardization of the entire process from primary sample collection to RNA analy

21、sis is needed. Studies have been undertaken to determine the important influencing factors. This Technical Specification draws upon such work to codify and standardize the steps for frozen tissue with regard to RNA analysis in what is referred to as the preanalytical phase. PD CEN/TS 16826-1:2015 CE

22、N/TS 16826-1:2015 (E) 5 1 Scope This Technical Specification gives recommendations for the handling, documentation and processing of frozen tissue specimens intended for RNA analysis during the preanalytical phase before a molecular assay is performed. This Technical Specification is applicable to m

23、olecular in vitro diagnostic examinations (e.g., in vitro diagnostic laboratories, laboratory customers, developers and manufacturers of in vitro diagnostics, institutions and commercial organisations performing biomedical research, biobanks, and regulatory authorities). RNA profiles in tissues can

24、change significantly before and after collection and can change differently in tissues from different donors / patients. Therefore, it is essential to take special measures to minimize the described profile changes and modifications within the tissue for subsequent RNA analysis. Tissues that have un

25、dergone chemical stabilisation pre-treatment before freezing are not covered in this document. 2 Normative references The following documents, in whole or in part, are normatively referenced in this document and are indispensable for its application. For dated references, only the edition cited appl

26、ies. For undated references, the latest edition of the referenced document (including any amendments) applies. EN ISO 15189:2012, Medical laboratories Requirements for quality and competence (ISO 15189:2012, Corrected version 2014-08-15) ISO 15190, Medical laboratories Requirements for safety 3 Term

27、s and definitions For the purposes of this document, the terms and definitions given in EN ISO 15189:2012 and the following apply. 3.1 ambient temperature unregulated temperature of the surrounding air 3.2 analytical phase processes that start with the isolated analyte and include all kinds of param

28、eter testing or chemical manipulation for quantitative or qualitative analysis 3.3 cold ischemia condition after removal of the tissue from the body until its stabilization or fixation 3.4 pre-examination processes preanalytical phase preanalytical workflow processes that start, in chronological ord

29、er, from the clinicians request and include the examination request, preparation and identification of the patient, surgical procedure, collection of the primary sample(s), temporary storage, transportation to and within the analytical laboratory, aliquoting, retrieval, isolation of analytes, and en

30、d when the analytical examination begins PD CEN/TS 16826-1:2015 CEN/TS 16826-1:2015 (E) 6 SOURCE: EN ISO 15189:2012, definition 3.15, modified An additional term was added and more details were included. Note 1 to entry: The preanalytical phase may include preparative processes that may influence th

31、e outcome of the intended examination. 3.5 primary sample specimen discrete portion of a body fluid, breath, hair or tissue taken for examination, study or analysis of one or more quantities or properties assumed to apply for the whole SOURCE: EN ISO 15189:2012, 3.16, modified The term and definitio

32、n is used here without the original notes. 3.6 quantitative RNA profile RNA profile amounts of the individual RNA molecules that are present in a sample and that can be measured in the absence of any losses, inhibition and interference 3.7 RNA ribonucleic acid polymer of ribonucleotides occurring in

33、 a double-stranded or single-stranded form SOURCE: EN ISO 22174:2005, 3.1.3 3.8 room temperature temperature which is defined as 18 C to 25 C for the purposes of this document 3.9 sample one or more parts taken from a primary sample SOURCE: EN ISO 15189:2012, 3.24, modified The example was not taken

34、 over. 3.10 stability ability of a sample material, when stored under specified conditions, to maintain a stated property value within specified limits for a specified period of time SOURCE: ISO Guide 30:1992, 2.7 Note 1 to entry: The measured constituent for the purpose of this document is RNA. 3.1

35、1 warm ischemia warm Ischemia is the condition where the tissue is deprived of its normal blood supply containing oxygen and nutrients while the tissue is at body temperature 4 General considerations For general statements on primary sample collection and handling (including avoidance of cross conta

36、minations) see EN ISO 15189:2012, 5.4.4, 5.2.6. Consumables including kits shall be verified before use in examination (see EN ISO 15189:2012, 5.3.2.3); EN ISO 15189:2012, 5.5.1.2 and 5.5.1.3 can also apply. PD CEN/TS 16826-1:2015 CEN/TS 16826-1:2015 (E) 7 As all steps of a diagnostic workflow can i

37、nfluence the final analytical performance, the entire workflow comprising the preanalytical steps, including information on biomolecule stability and storage conditions, and analytical steps should be verified and validated (see EN ISO 15189). The stability of the specific quantitative RNA profile(s

38、) of interest should be investigated throughout the entire preanalytical workflow prior to the development and implementation of an analytical test. Before tissues stabilized by freezing, quantitative RNA profile can change e.g., by gene induction, gene down regulation and RNA degradation. These eff

39、ects depend on the duration of warm and cold ischemia and the ambient temperature before freezing. In addition, the described effects can vary in tissues from different donors / patients. Generally, the longer the warm and cold ischemia times and the higher the ambient temperature before freezing th

40、e tissue specimen, the higher is the risk that changes in the RNA profile can occur. NOTE Intraoperative warm ischemia can result in more pronounced changes of RNA profiles than during postoperative cold ischemia. RNA profiles can also vary, depending on the origin and type of tissue, the underlying

41、 disease, the surgical procedure, the drug regime, and drugs administered for anaesthesia or treatment of concomitant disease and on the different environmental conditions after the tissue removal from the body. As warm ischemia cannot be easily standardized, its time and duration should be document

42、ed. When it is not possible to avoid cold ischemia, its time of onset, duration shall be documented and temperatures of the specimen transport containers surroundings should be documented. Where the specimen is transported to another facility for freezing, the transport duration shall be documented

43、and the ambient conditions should also be documented. Safety regulations on transport and handling shall be considered (see EN ISO 15189:2012, 5.2.3 and 5.4.5 and ISO 15190). During the whole preanalytical workflow precautions shall be taken to avoid cross contamination between different samples. If

44、 a commercial product is not used in accordance with the manufacturers instructions, responsibility for its use and performance lies with the user. 5 Outside the laboratory 5.1 Primary tissue collection manual 5.1.1 Information about the primary sample donor The documentation should include, but is

45、not limited to: a) the primary donor / patient ID, which can be in the form of a code; b) the health status of the primary sample donor (e.g., healthy, disease type, concomitant disease); c) the information about routine medical treatment and special treatment prior to tissue collection (e.g., anaes

46、thetics, medications, surgical or diagnostic procedures (e.g., biopsy device used for the collection); d) the start of ischemia within the body (warm ischemia) by documenting the ischemia-relevant vessel ligation/clamping time point (usually arterial clamping time). 5.1.2 Information on the primary

47、tissue sample The documentation shall include, but is not limited to: PD CEN/TS 16826-1:2015 CEN/TS 16826-1:2015 (E) 8 a) the time point when tissue is removed from the body; b) the description of tissue type, tissue condition (e.g., diseased, unaffected by the disease) and organ tissue of origin, i

48、ncluding references to any marking applied in the operating theatre made by surgeon, radiologist or pathologist; c) the documentation steps described under 6.3, if freezing is performed outside the laboratory. 5.1.3 Information on the primary tissue sample processing The following steps shall be per

49、formed: 1. the documentation of any additions or modifications to the primary sample after removal from the body (e.g., labelling for the orientation of the specimen (e.g., ink-marking, stitches), incision(s); 2. the selection and use of transport containers and packages (e.g., cooling box, box for storing and transportation, vacuum packaging); 3. the selection and use of stabilisation procedures (e.g., cooling methods) for transport; NOTE 1 Accidentally freezing and thawing the tissue (e.g., by using cool packs in a wrong manner) will lead to