ASTM E1873-04

Standard Guide for Detection of Nucleic Acid Sequences by the Polymerase Chain Reaction Technique

SCOPE
1.1 This guide covers guidelines, recommendations, basic considerations, criteria, and principles to be employed when developing, utilizing, or assessing PCR procedures and specific protocols for the amplification and detection of nucleic acid sequences. This guide is not intended to be a standard procedure with a list of requirements for PCR detection of nucleic acids. This guide is intended to provide information that will assist the user in obtaining quality and reliable data.
1.2 Nucleic acid sequences that can be amplified by PCR include DNA, as well as RNA sequences; RNA sequences are suitable targets for PCR following reverse transcription of the RNA to complementary DNA (cDNA). This type of amplification technique is often called reverse transcription-PCR (RT-PCR).
1.3 This guide has been developed for use in any molecular biology/biotechnology laboratory. This includes, but is not limited to, laboratories that specialize in the diagnosis of human, animal, plant, or bacterial diseases.
1.4 This guide conveys the general procedural terminology of PCR technology used for the detection of nucleic acids.
1.5 This guide is a general one; it does not cover the additional guidance that would be needed for specific applications, for example, for the PCR detection of nucleic acid sequences of specific microorganisms.
1.6 This guide does not cover details of the various methods that can be utilized to identify PCR-amplified DNA sequences.
1.7 This guide does not cover specific variations of the basic PCR or RT-PCR technology (for example, quantitative PCR, real-time PCR, multiplex PCR, and in situ PCR), and it does not cover details of instrument calibration.
1.8 Warning—Laboratory work involving certain clinical specimens and microorganisms can be hazardous to personnel. Warning—Biosafety level 2 (or higher) facilities are recommended for biohazard work (8). Safety guidelines should be adhered to in accordance with NCCLS M29-A2 and other recommendations (8).

General Information

• Status: Historical
• Publication Date: 30-Jun-2004
• ICS: 07.100.99 - Other standards related to microbiology
• Technical Committee: E48 - Bioenergy and Industrial Chemicals from Biomass

Relations

• Replaces: ASTM E1873-97(2002) - Standard Guide for Detection of Nucleic Acid Sequences by the Polymerase Chain Reaction Technique
• Effective Date: 01-Jul-2004
• Replaced By: ASTM E1873-06 - Standard Guide for Detection of Nucleic Acid Sequences by the Polymerase Chain Reaction Technique (Withdrawn 2014)
• Effective Date: 01-Nov-2006
• Referred By: ASTM E2800-11(2017) - Standard Practice for Characterization of <emph type="ital">Bacillus</emph> Spore Suspensions for Reference Materials
• Effective Date: 01-Jul-2004

Standards Content (Sample)

NOTICE: This standard has either been superseded and replaced by a new version or withdrawn.
Contact ASTM International (www.astm.org) for the latest information
Designation:E1873–04
Standard Guide for
Detection of Nucleic Acid Sequences by the Polymerase
1
Chain Reaction Technique
This standard is issued under the fixed designation E1873; the number immediately following the designation indicates the year of
original adoption or, in the case of revision, the year of last revision.Anumber in parentheses indicates the year of last reapproval.A
superscript epsilon (e) indicates an editorial change since the last revision or reapproval.
INTRODUCTION
This guide applies to the detection of deoxyribonucleic acid (DNA) or ribonucleic acid (RNA)
sequences by the polymerase chain reaction (PCR) technique. The PCR is used as a tool in many
molecular biology laboratory settings and for diverse reasons, for example, for amplification and
detection of nucleic acid sequences and for DNAsequencing. There is an abundance of publications
addressing laboratory procedures and specific protocols for various applications. The field of PCR is
advancingsorapidlythatitisnecessarytofrequentlymodifyandupdatetheseproceduresandspecific
protocols. This guide consists of guidelines, recommendations, basic considerations, criteria, and
principles that should be employed when developing, utilizing, or assessing PCR procedures and
specific protocols for the amplification and detection of nucleic acid sequences.
This guide was developed by Subcommittee E48.02 on Characterization and Identification of
Biological Systems in collaboration with DIN (German Institute for Standardization) Committee E9
onSerodiagnosisofInfectiousDiseasesandDiseasesoftheImmuneSystem,DepartmentforMedical
Standards (NAMed).
This guide assumes a basic knowledge of molecular biology. It assumes the availability of basic
2
references in PCR for general procedures (see Refs 1-7) and the ability to search the literature for
target-specific protocols.
1. Scope 1.3 This guide has been developed for use in any molecular
biology/biotechnology laboratory. This includes, but is not
1.1 This guide covers guidelines, recommendations, basic
limited to, laboratories that specialize in the diagnosis of
considerations, criteria, and principles to be employed when
human, animal, plant, or bacterial diseases.
developing,utilizing,orassessingPCRproceduresandspecific
1.4 This guide conveys the general procedural terminology
protocols for the amplification and detection of nucleic acid
of PCR technology used for the detection of nucleic acids.
sequences. This guide is not intended to be a standard
1.5 This guide is a general one; it does not cover the
procedure with a list of requirements for PCR detection of
additional guidance that would be needed for specific applica-
nucleic acids. This guide is intended to provide information
tions, for example, for the PCR detection of nucleic acid
that will assist the user in obtaining quality and reliable data.
sequences of specific microorganisms.
1.2 Nucleic acid sequences that can be amplified by PCR
1.6 Thisguidedoesnotcoverdetailsofthevariousmethods
include DNA, as well as RNAsequences; RNAsequences are
thatcanbeutilizedtoidentifyPCR-amplifiedDNAsequences.
suitable targets for PCR following reverse transcription of the
1.7 Thisguidedoesnotcoverspecificvariationsofthebasic
RNA to complementary DNA (cDNA). This type of amplifi-
PCR or RT-PCR technology (for example, quantitative PCR,
cation technique is often called reverse transcription-PCR
real-time PCR, multiplex PCR, and in situ PCR), and it does
(RT-PCR).
not cover details of instrument calibration.
1.8 Warning—Laboratory work involving certain clinical
1
ThisguideisunderthejurisdictionofASTMCommitteeE48onBiotechnology
specimens and microorganisms can be hazardous to personnel.
and is the direct responsibility of Subcommittee E48.02 on Characterization and
Warning—Biosafety level 2 (or higher) facilities are recom-
Identification of Biological Systems.
CurrenteditionapprovedJuly1,2004.PublishedJuly2004.Originallyapproved mended for biohazard work (8). Safety guidelines should be
in 1997. Last previous edition approved in 2002 as E1873–97(2002).
adhered to in accordance with NCCLS M29-A2 and other
2
Theboldfacenumbersinparenthesesrefertothelistofreferencesattheendof
recommendations (8).
this standard.
Copyright © ASTM International, 100 Barr Harbor Drive, PO Box C700, West Conshohocken, PA 19428-2959, United States.
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E1873–04
2. Referenced Documents ficity, sensitivity, and precision of the amplification reaction.
There are several ways to achieve hot-start PCR. All methods
2.1 NCCLS Standards:
involve withholding a critical component (for example, poly-
C24-A2 Statistical Quality Control for Quantitative Mea-
++
merase or Mg ) during the reaction setup at room tempera-
surements: Principles and Definitions; Approved
3
ture.Reactiontubesarethenheatedto
...