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ASTM E1531-00(2006)

(Practice)

Standard Practice for Detection of Mycoplasma Contamination of Cell Cultures by Growth on Agarose Medium (Withdrawn 2014)

Standard Practice for Detection of Mycoplasma Contamination of Cell Cultures by Growth on Agarose Medium (Withdrawn 2014)

ABSTRACT
This practice covers the procedures used for detection of mycoplasma contamination of cell cultures by growth on agarose medium. This practice does not cover identification of mycoplasma and indirect methods for detection of mycoplasma. This practice will not detect cultivar strains of Mycoplasma hyorhinis nor intended for use in detection of mycoplasma contamination in sera, culture media, vaccines, or other systems. The practice involves DM-1 solid medium preparation, quality control, and mycoplasma isolation.
SCOPE
1.1 This practice covers the procedures used for detection of mycoplasma contamination by direct microbiological culture.
1.2 This practice does not cover indirect methods for detection of mycoplasma such as DNA staining, biochemical detection, or genetic probes.
1.3 This practice does not cover methods for identification of mycoplasma organisms.
1.4 This practice will not detect cultivar strains (1) of Mycoplasma hyorhinis.
1.5 This practice is not intended for use in detection of mycoplasma contamination in sera, culture media, vaccines, or other systems.
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.
WITHDRAWN RATIONALE
This practice covers the procedures used for detection of mycoplasma contamination by direct microbiological culture.
Formerly under the jurisdiction of Committee E55 on Manufacture of Pharmaceutical Products, this practice was withdrawn in August 2014. This standard was withdrawn without replacement due to its limited use by the industry.

General Information

Status
Withdrawn
Publication Date
31-Oct-2006
Withdrawal Date
04-Aug-2014
ICS
07.100.01 - Microbiology in general
Technical Committee
E55 - Manufacture of Pharmaceutical and Biopharmaceutical Products
Drafting Committee
E55.04 - General Biopharmaceutical Standards
Current Stage
Ref Project

Relations

Replaces
ASTM E1531-00 - Standard Practice for Detection of Mycoplasma Contamination of Cell Cultures by Growth on Agarose Medium
Effective Date
01-Nov-2006
Referred By
ASTM E2363-23 - Standard Terminology Relating to Manufacturing of Pharmaceutical and Biopharmaceutical Products in the Pharmaceutical and Biopharmaceutical Industry
Effective Date
01-Nov-2006

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ASTM E1531-00(2006) - Standard Practice for Detection of Mycoplasma Contamination of Cell Cultures by Growth on Agarose Medium (Withdrawn 2014)ASTM E1531-00(2006) - Standard Practice for Detection of Mycoplasma Contamination of Cell Cultures by Growth on Agarose Medium (Withdrawn 2014)
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ASTM E1531-00(2006) - Standard Practice for Detection of Mycoplasma Contamination of Cell Cultures by Growth on Agarose Medium (Withdrawn 2014)
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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: E1531 − 00(Reapproved 2006)
Standard Practice for
Detection of Mycoplasma Contamination of Cell Cultures by
Growth on Agarose Medium
This standard is issued under the fixed designation E1531; 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 (´) indicates an editorial change since the last revision or reapproval.
1. Scope 3.1.2 indirect detection of mycoplasma, n—detection of
mycoplasma by DNA staining or any method other than
1.1 Thispracticecoverstheproceduresusedfordetectionof
cultivation.
mycoplasma contamination by direct microbiological culture.
3.1.3 mycoplasma (Mollicute), n—smallest prokaryotes ca-
1.2 This practice does not cover indirect methods for
pable of self replication.
detection of mycoplasma such as DNA staining, biochemical
detection, or genetic probes.
4. Significance and Use
1.3 This practice does not cover methods for identification
4.1 The demonstration of characteristic colonial growth on
of mycoplasma organisms.
axenic solid medium is a sensitive and specific method to
detect mycoplasma infection of cell cultures and it is the
1.4 This practice will not detect cultivar α strains (1) of
Mycoplasma hyorhinis. standard detection method (2).
4.2 When mycoplasmas contaminate cell cultures they usu-
1.5 This practice is not intended for use in detection of
mycoplasma contamination in sera, culture media, vaccines, or allygrowtohightiter(10 colonyformingunits/mL)andwhen
inoculatedontoagarmediumtheyproduceabundantandeasily
other systems.
detectable growth (3).
1.6 This standard does not purport to address all of the
safety concerns, if any, associated with its use. It is the 4.3 M. hyorhinis cultivar α strains do not grow on conven-
responsibility of the user of this standard to establish appro-
tional mycoplasma media (1) but require an indicator cell
priate safety and health practices and determine the applica- culture system to detect their presence (see Practice E1532).
bility of regulatory limitations prior to use.
Alternatively, a specialized axenic medium is suitable for
direct isolation of cultivar α from infected cell cultures (4).
2. Referenced Documents
4.4 Immunofluorescent procedures are used to identify my-
2.1 ASTM Standards:
coplasma isolates (5).
E1532 PracticeforDetectionofMycoplasmaContamination
5. DM-1 Solid Medium Preparation
of Cell Cultures by Use of Bisbenzamide DNA-Binding
Fluorochrome
5.1 Dissolve CMRL-1066 powder (CMRL-1066 powder
4,5
Formula No. 78–5156EF, packaged for 10L), in 5000 mL of
3. Terminology
distilled water. This is one-half the volume of water specified
5,6
3.1 Definitions:
on the package. Add 47.6 g HEPES, and 9.35 g NaCl.
3.1.1 direct mycoplasma detection, n—demonstration of
5.2 Adjust the pH to 7.3 and filter sterilize (450 nm). Store
characteristic colonial growth on axenic agar medium.
this 2X CMRL in the refrigerator in 500 mL amounts.
5,7 5,8
5.3 Dissolve 10.0 g of Myosate and 12 g of agarose in
400 mL of distilled water. Autoclave at 121°C for 15 minutes.
This practice is under the jurisdiction of ASTM Committee E55 on Manufac-
ture of Pharmaceutical Products and is the direct responsibility of Subcommittee
E55.04 on General Biopharmaceutical Standards.
Current edition approved Nov. 1, 2006. Published December 2006. Originally The sole source of supply of the apparatus known to the committee at this time
approved in 1993. Last previous edition approved in 2000 as E1531 – 00. DOI: is Life Technologies, Gaithersburg, MD.
10.1520/E1531-00R06. If you are aware of alternative suppliers, please provide this information to
The boldface numbers in parenthesis refer to the list of references at the end of ASTM International Headquarters. Your comments will receive careful consider-
this standard. ation at a meeting of the responsible technical committee, which you may attend.
3 6
For referenced ASTM standards, visit the ASTM website, www.astm.org, or The sole source of supply of the apparatus known to the committee at this time
contact ASTM Customer Service at service@astm.org. For Annual Book of ASTM is Research Organics, Cleveland, OH.
Standards volume information, refer to the standard’s Document Summary page on The sole source of supply of the apparatus known to the committee at this time
the ASTM website. is BBL Microbiology Systems, Cockeysville, MD.
Copyright © ASTM International, 100 Barr Harbor Drive, PO Box C700, West Conshohocken, P
...

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1.6 The values stated in SI units are to be regarded as standard. No other units of measurement are included in this standard.  
1.7 ASTM International takes no position respecting the validity of any patent rights asserted in connection with any item mentioned in this standard. Users of this standard are expressly advised that determination of the validity of any such patent rights, and the risk of infringement of such rights, are entirely their own responsibility.  
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5.1 Bacteria that exist in biofilms are phenotypically different from suspended cells of the same genotype. Research has shown that biofilm bacteria are more difficult to kill than suspended bacteria (5, 7). Laboratory biofilms are engineered in growth reactors designed to produce a specific biofilm type. Altering system parameters will correspondingly result in a change in the biofilm. For example, research has shown that biofilm grown under high shear is more difficult to kill than biofilm grown under low shear (5, 8). The purpose of this test method is to direct a user in the laboratory study of a Pseudomonas aeruginosa biofilm by clearly defining each system parameter. This test method will enable an investigator to grow, sample, and analyze a Pseudomonas aeruginosa biofilm grown under high shear. The biofilm generated in the CDC Biofilm Reactor is also suitable for efficacy testing. After the 48 h growth phase is complete, the user may add the treatment in situ or remove the coupons and treat them individually.
SCOPE
1.1 This test method specifies the operational parameters required to grow a reproducible (1)2 Pseudomonas aeruginosa ATCC 700888 biofilm under high shear. The resulting biofilm is representative of generalized situations where biofilm exists under high shear rather than being representative of one particular environment.  
1.2 This test method uses the Centers for Disease Control and Prevention (CDC) Biofilm Reactor. The CDC Biofilm Reactor is a continuously stirred tank reactor (CSTR) with high wall shear. Although it was originally designed to model a potable water system for the evaluation of Legionella pneumophila (2), the reactor is versatile and may also be used for growing and/or characterizing biofilm of varying species (3-5).  
1.3 This test method describes how to sample and analyze biofilm for viable cells. Biofilm population density is recorded as log10 colony forming units per surface area.  
1.4 Basic microbiology training is required to perform this test method.  
1.5 The values stated in SI units are to be regarded as standard. No other units of measurement are included in this standard.  
1.6 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.  
1.7 This international standard was developed in accordance with internationally recognized principles on standardization established in the Decision on Principles for the Development of International Standards, Guides and Recommendations issued by the World Trade Organization Technical Barriers to Trade (TBT) Committee.

  • Standard
    7 pages
    English language
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  • Standard
    7 pages
    English language
    sale 15% off