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ASTM_E_1873_-_06.pdf

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1、Designation:E1873 06Standard Guide forDetection of Nucleic Acid Sequences by the PolymeraseChain Reaction Technique1This standard is issued under the fixed designation E1873;the number immediately following the designation indicates the year oforiginal adoption or,in the case of revision,the year of

2、 last revision.A number in parentheses indicates the year of last reapproval.Asuperscript epsilon()indicates an editorial change since the last revision or reapproval.INTRODUCTIONThis guide applies to the detection of deoxyribonucleic acid(DNA)or ribonucleic acid(RNA)sequences by the polymerase chai

3、n reaction(PCR)technique.The PCR is used as a tool in manymolecular biology laboratory settings and for diverse reasons,for example,for amplification anddetection of nucleic acid sequences.There is an abundance of publications addressing laboratoryprocedures and specific protocols for various applic

4、ations.The field of PCR is advancing so rapidlythat it is necessary to frequently modify and update these procedures and specific protocols.This guideconsists of guidelines,recommendations,basic considerations,criteria,and principles that should beemployed when developing,utilizing,or assessing PCR

5、procedures and specific protocols for theamplification and detection of nucleic acid sequences.This guide was developed by Subcommittee E48.02 on Characterization and Identification ofBiological Systems.The 1997 edition of this guide was developed in collaboration with DIN(GermanInstitute for Standa

6、rdization)Committee E9 on Serodiagnosis of Infectious Diseases and Diseases ofthe Immune System,Department for Medical Standards(NAMed).This guide assumes a basic knowledge of molecular biology.It assumes the availability of basicreferences in PCR for general procedures(see Refs 1-7)2and the ability

7、 to search the literature fortarget-specific protocols.1.Scope1.1 This guide covers guidelines,recommendations,basicconsiderations,criteria,and principles to be employed whendeveloping,utilizing,or assessing PCR procedures and specificprotocols for the amplification and detection of nucleic acidsequ

8、ences.This guide is not intended to be a standardprocedure with a list of requirements for PCR detection ofnucleic acids.This guide is intended to provide informationthat will assist the user in obtaining quality and reliable data.1.2 Nucleic acid targets for PCR include DNA,as well asRNA;RNA sequen

9、ces are suitable targets for PCR followingreverse transcription of the RNA to complementary DNA(cDNA).This type of amplification technique is known asreverse transcription-PCR(RT-PCR).1.3 This guide has been developed for use in any molecularbiology/biotechnology laboratory.This includes,but is notl

10、imited to,laboratories that specialize in the diagnosis ofhuman,animal,plant,or bacterial diseases.1.4 This guide conveys the general procedural terminologyof PCR technology used for the detection of nucleic acids.1.5 This guide is general;it does not cover the additionalguidance that would be neede

11、d for specific applications,forexample,for the PCR detection of nucleic acid sequences ofspecific microorganisms.1.6 This guide does not cover details of the various methodsthat can be utilized to identify PCR-amplified DNA sequences.1.7 This guide does not cover specific variations of the basicPCR

12、or RT-PCR technology(for example,quantitative PCR,real-time PCR,multiplex PCR,and in situ PCR),and it doesnot cover details of instrument calibration.1.8 WarningLaboratory work involving certain clinicalspecimens and microorganisms can be hazardous to personnel.WarningBiosafety level 2(or higher)fac

13、ilities are recom-mended for biohazard work(8).Safety guidelines should beadhered to in accordance with CLSI M29-A2 and otherrecommendations(8).1This guide is under the jurisdiction of ASTM Committee E55 on Manufactureof Pharmaceutical Products and is the direct responsibility of Subcommittee E55.04

14、on General Biopharmaceutical Standards.Current edition approved Nov.1,2006.Published November 2006.Originallyapproved in 1997.Last previous edition approved in 2004 as E1873 04.DOI:10.1520/E1873-06.2The boldface numbers in parentheses refer to the list of references at the end ofthis standard.1Copyr

15、ight ASTM International,100 Barr Harbor Drive,PO Box C700,West Conshohocken,PA 19428-2959,United States.Copyright ASTM International Provided by IHS under license with ASTM Licensee=Ohio State University/5967164005,User=ahmadi,rozitaNot for Resale,03/26/2012 04:38:14 MDTNo reproduction or networking

16、 permitted without license from IHS-,-,-2.Referenced Documents2.1CLSI Standards:3C24-A2 Statistical Quality Control for Quantitative Mea-surements:PrinciplesandDefinitions;ApprovedGuidelineSecond Edition(1999)M29-A2 Protection of Laboratory Workers from Occupa-tionally Acquired InfectionsSecond Edition;ApprovedGuideline(2001)GP5-A2 Clinical Laboratory Waste Management;ApprovedGuidelineSecond Edition(2002)MM3-A2 Molecular Diagnostic Methods for InfectiousDiseases;Approved GuidelineSecond Edition(

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