1.1 本规程用于量化液体或固体去污剂对 芽孢杆菌 孢子在多孔和非多孔材料制成的试样表面干燥。这种做法可以区分去污混合物中的杀菌和抑菌化学品。这一点很重要，因为许多去污剂同时含有活性化合物和高浓度的抑菌表面活性剂。所有测试样品在测试当天直接与预中和对照品、未接种的负生长对照品和溶液对照品进行比较，以增加灭活数据的实际可信度。 1.2 该程序只能由受过微生物学技术培训、熟悉抗菌剂（杀孢剂）和抗菌产品使用说明书的人员执行。 1.3 以国际单位制表示的数值应视为标准值。本标准不包括其他计量单位。 1.4 本标准可能涉及危险材料、操作和设备。本标准并非旨在解决与其使用相关的所有安全问题（如有）。本标准的用户有责任在使用前制定适当的安全、健康和环境实践，并确定监管限制的适用性。 1.5 本国际标准是根据世界贸易组织技术性贸易壁垒（TBT）委员会发布的《关于制定国际标准、指南和建议的原则的决定》中确立的国际公认标准化原则制定的。 ====意义和用途====== 5.1 本规程可用于评估任何成分的试片材料，只要试片足够小，可以安装在中提到的过滤器单元内 4.1 . 5.2 该实践定义了定量、可扩展、快速、敏感和安全的程序，同时最大限度地减少劳动力并解决统计置信度问题 ( 1. , 2. ) . 5.2.1 定量- 通过稀释平板法确定每个试片的孢子总数，并在提取管中对试片上剩余的所有孢子进行活性分析，以确保所有孢子均已被计算在内。 5.2.2 统计置信度- 使用五种独立的孢子接种剂进行统计分析 n 第5页。 5.2.3 灵敏度- 允许完全检测接种在试片上的所有可培养孢子，包括仍然附着在试片上的孢子。 5.2.3.1 检测极限取决于完全成熟孢子在提取介质（1%胰蛋白酶大豆肉汤、100 mM L-丙氨酸、1 mM肌苷、0.05%吐温80）和/或胰蛋白酶大豆琼脂存在下萌发、生长和分裂的培养能力。 5.2.3.2 结果见参考文献 ( 1. , 3. ) （以及目前尚未公布的结果）表明，这些培养基，结合本文所述的测试温度和条件，将产生具有高水平实际置信度的结果，用于检测可培养物 芽孢杆菌 孢子。 5.2.4 安全- 接种的试片包含在过滤单元内。 5.2.5 测试的简单性- 测试和提取在同一过滤装置中进行，以尽量减少试样处理步骤。 5.2.6 可扩展且快速- 两名技术人员最多可在1小时内处理36个样品；六名技术人员在5小时内总共处理了300个样品 ( 1. , 2. ) . 5.2.7 广泛应用于众多领域 芽孢杆菌 物种和菌株。该方法也已被修改，并用于植物细菌和病毒 ( 1. , 2. ) .

1.1 This practice is used to quantify the efficacy of liquid or solid decontaminants on Bacillus spores dried on the surface of coupons made from porous and non-porous materials. This practice can distinguish between bactericidal and bacteriostatic chemicals within decontamination mixtures. This is important because many decontaminants contain both reactive compounds and high concentrations of bacteriostatic surfactants. All test samples are directly compared to pre-neutralized controls, un-inoculated negative growth controls, and solution controls on the same day as the test in order to increase practical confidence in the inactivation data. 1.2 This procedure should be performed only by those trained in microbiological techniques, are familiar with antimicrobial (sporicidal) agents and the application instructions of the antimicrobial products. 1.3 The values stated in SI units are to be regarded as standard. No other units of measurement are included in this standard. 1.4 This standard may involve hazardous materials, operations, and equipment. 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, health, and environmental practices and determine the applicability of regulatory limitations prior to use. 1.5 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. ====== Significance And Use ====== 5.1 The practice can be used to evaluate coupon materials of any composition, insofar as the coupon can be small enough to fit inside filter units mentioned in 4.1 . 5.2 This practice defines procedures that are quantitative, scalable, rapid, sensitive, and safe, while minimizing labor and addressing statistical confidence ( 1 , 2 ) . 5.2.1 Quantitative— The total number of spores per coupon is determined by dilution-plating, and all spores remaining on the coupon are assayed for activity in the extraction tube to provide confidence that all the spores were accounted for. 5.2.2 Statistical Confidence— The use of five independent preparations of spore inocula for a statistical n of 5. 5.2.3 Sensitivity— Allows for complete detection of all culturable spores inoculated on a coupon, including the spores that remain attached to the coupon. 5.2.3.1 The limit of detection is dependent on the culturability of fully matured spores to germinate, outgrow and divide in the presence of the extraction medium (1% tryptic soy broth, 100 mM L-Alanine, 1 mM inosine, 0.05% Tween 80) and/or on tryptic soy agar. 5.2.3.2 Results presented in Refs ( 1 , 3 ) (and currently unpublished results) indicate that these media, combined with the test temperatures and conditions described herein will generate results with a high level of practical confidence for detecting culturable Bacillus spores. 5.2.4 Safety— Inoculated coupons are contained within filter units. 5.2.5 Simplicity of Testing— Tests and extractions are performed in the same filter unit to minimize coupon handling steps. 5.2.6 Scalable and Rapid— A maximum of 36 samples can be processed in 1 h by two technicians; a total of 300 samples have been processed by six technicians in 5 h ( 1 , 2 ) . 5.2.7 Wide application for numerous Bacillus species and strains. The method has also been modified and used for vegetative bacteria and viruses as well ( 1 , 2 ) .