1.1 本实践侧重于两个基本测量来表征 芽孢杆菌 参考材料，使用营养培养基上菌落的生长和相衬显微镜测定孢子质量和同质性来计数孢子。有关高级表征方法的更多信息，请参阅 附录X1 . 1.2 本文件将为用户提供测量方法的建议，以及应采用的细节和条件，以确保可靠和高可靠性- 获得了质量数据。该实践将有助于确保以统一的方式报告从表征中获得的结果。这将允许其他人复制测量结果，并便于比较不同批次的数据 芽孢杆菌 用作参考材料的孢子悬浮液。需要注意的是 芽孢杆菌 物种是一个异质群体，其生长和产孢的具体要求可能不同。鼓励本规程的使用者查阅文献，以获取有关该物种的具体信息 芽孢杆菌 他们使用的细菌 ( 1. ) . 2. 1.3 本标准规程不提供未知细菌种类的鉴定指南。识别 芽孢杆菌 传统上，物种是基于菌落形态、选择性培养基上的生长和生化测试，但最近的核酸技术已经能够基于16S DNA序列相似性对该群体进行系统发育分析 ( 1. ) . 1.4 一些 芽孢杆菌 spp.对人类和动物具有致病性，建议用户遵守安全的实验室程序和操作规程，以处理来自这些物种的孢子 ( 2. ) . 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和健康实践，并确定监管限制的适用性 ( 2. ) . 1.5 本实践假设具备微生物学和分子生物学的基本知识，并可访问引用的参考文献。 1.6 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。 1.7 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 用作参考材料的孢子特性的标准实践对于确保检测设备和实验室仪器性能测试的统一基础非常重要。 芽孢杆菌 孢子悬浮液可用于多种用途，包括测试环境采样技术、灭活方法、去污方法和基础研究。 5.2 本规程适用于以下设备的制造商和最终用户: 芽孢杆菌 孢子悬浮液。特性测量的结果在分析报告（ROA）中给出。ROA应提供有关测量技术的足够详细信息，以使客户能够复制测量结果，使他们能够确定孢子悬浮液的特性在运输和储存期间是否发生变化。 5.3 活孢子的计数和通过显微镜分析确定均匀性是标准物质最小表征所需的两种基本测量。相衬显微镜不需要染色来区分“相亮”休眠孢子与相暗孢子、深色营养细胞和团块。当孢子萌发时，在相位对比成像下，它们呈现相位暗 ( 5. ) . 由于缺乏营养，参考样品中萌发的孢子很快就会死亡。 保存样品时，防止孢子过早萌发很重要。本标准实施规程包括这些测量的重要步骤，并包括高级测量的指南。为表征孢子悬浮液的先进技术提供了额外的指导，这些技术可用于提供更高水平的表征 芽孢杆菌 孢子参考样品。 5.4 除了本规程中涵盖的基本测量外，还应确定用于其预期应用的孢子的特定特性，例如对消毒剂过程的敏感性。 有关孢子特性测量的更多信息，请参阅附录。

1.1 This practice is focused on two basic measurements to characterize Bacillus reference materials, the enumeration of spores using growth of colonies on nutrient media and using phase contrast microscopy to determine spore quality and homogeneity. Additional information on advanced methods for characterization is provided in Appendix X1 . 1.2 This document will provide the user with recommendations for measurement methods, and the details and conditions that should be employed to ensure reliable and high-quality data are obtained. The practice will help ensure that results obtained from the characterization are reported in a uniform manner. This will allow others to replicate the measurements and facilitate the comparison of different lots of Bacillus spore suspensions used as reference materials. It is important to note that the Bacillus species are a heterogeneous group and their specific requirements for growth and sporulation may vary. Users of this practice are encouraged to consult the literature for specific information on the species of Bacillus bacteria they are using ( 1 ) . 2 1.3 This standard practice does not provide guidance for the identification of unknown species of bacteria. The identification of Bacillus species has been traditionally based on colony morphology, growth on selective media, and biochemical tests, but more recently nucleic acid technologies have enabled the phylogenetic analysis of this group based on 16S DNA sequence similarities ( 1 ) . 1.4 Some Bacillus spp. are pathogenic to humans and animals and the user is advised to adhere to safe laboratory procedures and practices for handling spores from these species ( 2 ) . 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 ) . 1.5 This practice assumes a basic knowledge of microbiology and molecular biology and access to the cited references. 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 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 Standard practices for the characterization of spores used as reference materials are important to ensure a uniform basis for testing the performance of detection devices and laboratory instruments. Bacillus spore suspensions can be used for a large variety of purposes including testing environmental sampling techniques, inactivation methods, decontamination methods and basic research. 5.2 The practice is intended for both manufacturers and end users of Bacillus spore suspensions. The results of the characterization measurements are presented in a report of analysis (ROA). The ROA should provide sufficient detail about the measurement technique to enable the customers to replicate the measurements, allowing them to determine if the properties of the spore suspension changed during shipping and storage. 5.3 The enumeration of the viable spores and determination of homogeneity by microscopic analysis are two basic measurements required for the minimal characterization of reference materials. Phase contrast microscopy does not require staining to distinguish the “phase bright” dormant spores from phase dark spores, dark vegetative cells and clumps. When spores germinate they appear phase dark under phase contrast imaging ( 5 ) . Germinated spores in a reference sample will soon die due to lack of nutrients. It is important in storing samples to prevent the premature germination of the spores. This standard practice includes the important steps for these measurements and includes guidance for advanced measurements. Additional guidance is given for advanced techniques to characterize spore suspensions that may be used to provide a higher level of characterized Bacillus spore reference samples. 5.4 The specific properties of the spores used for their intended application, such as susceptibility to disinfectant processes, should be determined in addition to the basic measurements covered in this practice. Additional information on the measurement of spore properties is located in the appendix.