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Significance and Use
5.1 The quantity and quality of the spores produced by this method meet the current requirements of the U.S. Environmental Protection Agency (EPA) to assess environmental surface disinfectants for label claims of sporicidal activity (4). The method is applicable to standard as well as clinically isolated toxigenic and non-toxigenic strains of C. difficile.
1.1 This test method describes the production and semipurification of C. difficile spores (also called endospores) primarily for use in testing the sporicidal activities of environmental surface disinfectants (Test Methods E2111and E2197); such spores can also be used to study their structure, chemistry and germination.
1.2 While the test method described is based on the use of 500-mL volumes of the liquid culture medium in an anaerobic incubator, anaerobic jars with smaller volumes of the same medium can also be used.
1.3 It is the responsibility of the investigator to determine whether Good Laboratory Practice (GLP) regulations are required and to follow them when appropriate (40 CFR, Part 160 for EPA submissions and 21 CFR; Part 58 for FDA submissions).
1.4 Warning—This standard may involve hazardous materials, chemicals, and microorganisms and should be performed only by persons with formal training in microbiology.
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.
2. Referenced Documents (purchase separately) The documents listed below are referenced within the subject standard but are not provided as part of the standard.
D1129 Terminology Relating to Water
E2111 Quantitative Carrier Test Method to Evaluate the Bactericidal, Fungicidal, Mycobactericidal, and Sporicidal Potencies of Liquid Chemicals
E2197 Quantitative Disk Carrier Test Method for Determining Bactericidal, Virucidal, Fungicidal, Mycobactericidal, and Sporicidal Activities of Chemicals
E2756 Terminology Relating to Antimicrobial and Antiviral Agents
E2839 Test Method for Production of Clostridium difficile Spores for Use in Efficacy Evaluation of Antimicrobial Agents
Federal Standards21 CFR, Part 58 Good Laboratory Practice for Nonclinical Laboratory Studies 40 CFR, Part 160 Good Laboratory Practice Standards
ICS Number Code 07.100.99 (Other standards related to microbiology)
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ASTM E2895-13, Standard Test Method for Producing High Titers of Viable and Semi-Purified Spores of Clostridium difficile using a Liquid Medium, ASTM International, West Conshohocken, PA, 2013, www.astm.orgBack to Top