ASTM E1286-00
(Guide)Standard Guide for Identification of Herpes Simplex Virus or its DNA
Standard Guide for Identification of Herpes Simplex Virus or its DNA
SCOPE
1.1 This guide covers laboratory characterization procedures sufficient to identify purified specimens of HSV types 1 and 2 (HSV-1 and HSV-2) or HSV-1 DNA and HSV-2 DNA used in biotechnology. For cases in which identification of HSV DNA specimens is required, the characterization criteria of 6.2 and 6.3 of this guide are sufficient.
1.2 This guide does not cover the identification of HSV in HSV-infected host cells. To apply this guide to such a case, it would first be necessary to isolate the virus from such samples using standard techniques of HSV purification. This guide does not cover characterization of segments of HSV DNA or of vectors containing HSV DNA segments.
1.3 This guide does not cover the specific methodology used in the identification characterization. It does not address the question of degree of purity required for herpesvirus preparations: this would vary depending on the particular biotechnology use of the virus.
1.4 Warning--Laboratory work involving herpes simplex viruses can be hazardous to personnel. Precaution: Biosafety 2 level facilities are recommended (1). Safety guidelines shall be adhered to according to NCCLS M29-T2 and other recommendations (1).
1.5 This standard does not purport to address all of the safety concerns, if any, associated with its use. It is the responsibility of the user of this standard to establish appropriate safety and health practices and determine the applicability of regulatory limitations prior to use.
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Designation: E 1286 – 00
Standard Guide for
Identification of Herpes Simplex Virus or Its DNA
This standard is issued under the fixed designation E 1286; the number immediately following the designation indicates the year of
original adoption or, in the case of revision, the year of last revision. A number 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 covers the identification of herpes simplex virus (HSV) or its DNA and was developed
by Subcommittee E48.02 on Characterization and Identification of Biological Systems. The objective
is to describe laboratory characterization procedures that would be sufficient to verify that a biological
preparation believed to contain primarily HSV (or HSV DNA) for use in any step of a biotechnology
process actually does contain this virus (or its DNA).
This guide assumes a basic knowledge of virology and molecular biology.
1. Scope priate safety and health practices and determine the applica-
bility of regulatory limitations prior to use.
1.1 This guide covers laboratory characterization proce-
dures sufficient to identify purified specimens of HSV types 1
2. Referenced Documents
and 2 (HSV-1 and HSV-2) or HSV-1 DNA and HSV-2 DNA
2.1 ASTM Standards:
used in biotechnology. For cases in which identification of
E 1873 Guide for Detection of Nucleic Sequences by the
HSV DNA specimens is required, the characterization criteria
Polymerase Chain Reaction Technique
of 6.2 and 6.3 of this guide are sufficient.
2.2 NCCLS Standards:
1.2 This guide does not cover the identification of HSV in
M29–T2 Protection of Laboratory Workers from Infectious
HSV-infected host cells. To apply this guide to such a case, it
DiseaseTransmitted by Blood, Body Fluids, andTissue—
would first be necessary to isolate the virus from such samples
Second Edition; Tentative Guideline
usingstandardtechniquesofHSVpurification.Thisguidedoes
not cover characterization of segments of HSV DNA or of
3. Terminology
vectors containing HSV DNA segments.
3.1 Basic polymerase chain reaction (PCR) definitions ap-
1.3 Thisguidedoesnotcoverthespecificmethodologyused
ply according to the general PCR Guide E 1873 (Section 3).
in the identification characterization. It does not address the
3.2 Definitions of Terms Specific to This Standard:
question of degree of purity required for herpesvirus prepara-
3.2.1 capsomere—a structural subunit of the outer protein
tions: this would vary depending on the particular biotechnol-
shell (capsid) of a virus consisting of protein monomers.
ogy use of the virus.
3.2.2 envelope—a layer of cell membrane-derived lipopro-
1.4 Warning—Laboratory work involving herpes simplex
tein that surrounds the protein coat (capsid) of some viruses.
viruses can be hazardous to personnel. Precaution: Biosafety
2 3.2.3 genome (of a virus)—the genetic material consisting
2 level facilities are recommended (1). Safety guidelines shall
of nucleic acid (RNA or DNA).
be adhered to according to NCCLS M29–T2 and other recom-
3.2.4 nucleocapsid—the outer protein coat or shell (capsid)
mendations (1).
of a virus plus its inner core of nucleic acid and proteins.
1.5 This standard does not purport to address all of the
3.2.5 plaque—a round, clear area in a layer of host cells
safety concerns, if any, associated with its use. It is the
caused by virus growth and resultant killing or lysis of the
responsibility of the user of this standard to establish appro-
cells.
ThisguideisunderthejurisdictionofASTMCommitteeE48onBiotechnology
and is the direct responsibility of Subcommittee E48.02 on Characterization and For referenced ASTM standards, visit the ASTM website, www.astm.org, or
Identification of Biological Systems. contact ASTM Customer Service at service@astm.org. For Annual Book of ASTM
Current edition approved September 10, 2000. Published December 2000. Standards volume information, refer to the standard’s Document Summary page on
Originally published as E 1286 – 89. Last previous edition E 1286 – 89 (1994). the ASTM website.
2 4
The boldface numbers in parentheses refer to a list of references at the end of Available from the National Committee for Clinical Laboratory Standards, 940
this guide. West Valley Road, Suite 1400, Wayne PA 19087.
Copyright © ASTM International, 100 Barr Harbor Drive, PO Box C700, West Conshohocken, PA 19428-2959, United States.
E1286–00
3.2.6 restriction endonuclease—a bacterial enzyme that 5.5 There are many uses of HSV or its DNA in basic and
cuts double-stranded DNA at positions consisting of specific applied biotechnology. Examples of applied uses include the
short sequences of nucleotides. preparation of DNA probes and monoclonal antibodies for in
vitro diagnostic testing and utilization of the virus in in vitro
4. Significance and Use
testing of antiviral substances.
4.1 This guide is intended for use in a biotechnology
laboratory whenever the necessity arises for identifying a
6. Characterization Criteria for Identification
biological preparation believed to contain primarily HSVor its
6.1 Immunological Evidence—Immunological evidence
DNA. The characterization criteria used for the identification
shall be provided such as demonstrating HSV envelope glyco-
shall be performed by an individual trained in molecular
protein antigen in viral-infected host cells by immunofluores-
virology.
cent (IF) or immunoperoxidase staining, orWestern blotting of
4.2 This guide is not meant to be used in a clinical
sodium dodecyl sulfate polyacrylamide gel electrophoresis
laboratory for the identification of HSV isolated from patient
(SDSPAGE)gels(forexample,seeRefs (3, 5 and 6)).Enzyme
specimens.
immunoassay(EIAorELISA)orradioimmunoassay(RIA)can
also be used to demonstrate the presence of HSV envelope
5. Background Information About Herpes Simplex Virus
glycoprotein (3, 5 and 6). HSV-1 can be differentiated from
5.1 Herpes simplex virus is a common human virus that can
HSV-2, and different strains of the viruses can be distin-
cause primary and recurrent infections of the skin and mucous
guished, by the use of appropriate monoclonal antibodies for
membranes (2-4). It has been classified by the International
immunofluorescence or EIA. Any one of a number of pub-
CommitteeonTaxonomyofVirusesas(a)Family:Herpesvirus
lished protocols can be used. It should be pointed out that,
group (Herpesviridae) and (b) Subfamily: Herpes simplex
although different species of herpes viruses have distinct
virus group (Alphaherpesvirinae) (2). There are two main
envelope glycoproteins, there are some shared antigenic deter-
immunologic variants of HSV, types 1 and 2 (HSV-1 and
minants (3).
HSV-2). They are officially known as human (alpha) herpes-
6.2 DNA Gel Electrophoresis—Gel electrophoresis patterns
virus 1 and human (alpha) herpesvirus 2 (2). HSV-1 has been
of restriction endonuclease fragments of isolated HSV DNA
isolated primarily from the oral cavity, eye, and skin vesicles
are distinctive. They can be used
...
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