1.1 本指南的目的是帮助非培养微生物试验的用户和生产者确定试验在处理不同类型样品方面的适用性，并评估结果的准确性。培养试验程序，如异养（标准）平板计数、最可能数（MPN）法和扩散平板计数，广泛用于微生物计数。然而，这些方法有其局限性，例如性能时间。此外，任何给定的培养试验方法通常只能回收样本中存在的全部活菌的一部分。正是这些局限性最近导致了各种非培养程序、测试试剂盒和仪器的营销。 1.2 培养试验方法基于样品中微有机物在指定生长条件下在指定生长介质中或在指定生长介质上增殖的能力来估计微生物种群密度。非文化测试方法试图通过测量不同的参数来提供相同或互补的信息。本指南旨在帮助研究人员评估用于测定微生物种群密度或活性的非培养方法的准确性和精密度。 1.3 众所周知，异养平板计数（HPC）不能回收产品或系统中存在的所有微生物 ( 1. , 2. ) . 2. 当在微生物种群的表征过程中出现此问题时，可能需要替代标准枚举程序，例如硫酸盐- 减少细菌。在其他情况下，可能需要使用化学方法来测量代谢衍生物的出现率、污染产品成分的利用率或微生物种群的遗传特征。在评估非培养试验时，使用这些替代标准程序可能是建立相关性的唯一方法。在这种情况下，本指南可以作为这些考虑的参考。 1.4 由于有太多类型的测试可以被视为非文化测试，因此不可能推荐具有统计分析的特定测试方案来评估测试。相反，本指南应帮助确定应考虑何种类型的测试来验证效用并确定非常规测试的局限性。 1.5 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。 1.6 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 非培养试验的生产者和潜在生产者应使用本指南来确定试验的准确性、选择性、特异性和精密度，如实践中所定义 E691 . 此类研究的结果应确定局限性，并指出非- 用于不同类型样品的培养试验，或两者兼有。指导 E1488 建议使用其他统计工具来评估拟议新测试方法的适用性和适用性。 5.2 非文化测试用户和潜在用户应使用本指南来评估非文化测试的结果，并与他们目前的方法进行比较。实践 D5245 和 D5465 应根据所采用的微生物方法进行审查。如果没有使用培养方法监测系统，则包括获取微生物专业知识的指南。 5.3 使用非培养测试可以减少确定系统微生物状态和检测培养测试未检测到的微生物所需的时间。 因此，非培养试验有助于提高微生物污染状况监测和诊断工作以及杀微生物剂性能评估的整体运行效率。 5.4 检测超过预定控制上限的微生物污染水平表明需要添加抗菌剂或采取其他纠正性维护措施。通过在比培养方法更短的时间内准确测定这一点，与将治疗推迟到培养结果可用相比，使用抗菌剂治疗可能会避免更严重的问题。如果抗菌治疗计划依赖于不准确的非培养测试，则会存在不必要的产品损失以及与抗菌剂选择不当或剂量不当相关的问题。 5.5 由于考虑了许多基于完全不同的化学和微生物原理的方法，因此不可能建立独特的设计并推荐用于进行比较的特定统计分析方法。只有在进行实验时才能提供应遵循的指南。还建议统计学家参与这项研究。

1.1 The purpose of this guide is to assist users and producers of non-culture microbiological tests in determining the applicability of the test for processing different types of samples and evaluating the accuracy of the results. Culture test procedures such as the Heterotrophic (Standard) Plate Count, the Most Probable Number (MPN) method and the Spread Plate Count are widely cited and accepted for the enumeration of microorganisms. However, these methods have their limitations, such as performance time. Moreover, any given culture test method typically recovers only a portion of the total viable microbes present in a sample. It is these limitations that have recently led to the marketing of a variety of non-culture procedures, test kits and instruments. 1.2 Culture test methods estimate microbial population densities based on the ability of mircoorganisms in a sample to proliferate in or on a specified growth medium, under specified growth conditions. Non-culture test methods attempt to provide the same or complimentary information through the measurement of a different parameter. This guide is designed to assist investigators in assessing the accuracy and precision of non-culture methods intended for the determination of microbial population densities or activities. 1.3 It is recognized that the Heterotrophic Plate Count (HPC) does not recover all microorganisms present in a product or a system ( 1 , 2 ) . 2 When this problem occurs during the characterization of a microbiological population, alternative standard enumeration procedures may be necessary, as in the case of sulfate-reducing bacteria. At other times, chemical methods that measure the rates of appearance of metabolic derivatives, the utilization of contaminated product components or genetic profile of the microbial population might be indicated. In evaluating non-culture tests, it is possible that the use of these alternative standard procedures might be the only means available for establishing correlation. In such cases, this guide can serve as a reference for those considerations. 1.4 Because there are so many types of tests that could be considered non-culture based, it is impossible to recommend a specific test protocol with statistical analyses for evaluating the tests. Instead, this guide should assist in determining what types of tests should be considered to verify the utility and identify the limitations of the nonconventional test. 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 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 This guide should be used by producers and potential producers of non-culture tests to determine the accuracy, selectivity, specificity, and precision of the tests, as defined in Practice E691 . Results of such studies should identify the limitations and indicate the utility or applicability of the non-culture test, or both, for use on different types of samples. Guide E1488 recommends other statistical tools for evaluating the suitability and applicability of proposed new test methods. 5.2 Non-culture test users and potential users should employ this guide to evaluate results of the non-culture test as compared to their present methods. Practices D5245 and D5465 should be reviewed in regards to the microbiological methods employed. If culture methods have not been used for monitoring the systems, then guidelines are included for obtaining microbiological expertise. 5.3 Utilization of a non-culture test can reduce the time required to determine the microbiological status of the system and detect microbe that are not detected by culture testing. Consequently, non-culture tests can contribute to the improvement in the overall operating efficiency of microbial contamination condition monitoring and diagnostic efforts, and microbicide performance evaluations. 5.4 Detecting microbial contamination levels that exceed predetermined upper control limits indicates the need for an addition of an antimicrobial agent or other corrective maintenance action. By accurately determining this in a shorter time period than is possible than by culture methods, treatment with antimicrobial agents may circumvent more serious problems than if the treatment were postponed until culture results were available. If the antimicrobial treatment program relied on an inaccurate non-culture test, then unnecessary loss of product and problems associated with inappropriate selection or improper dosing with antimicrobial agents would exist. 5.5 Since many methods based on entirely different chemical and microbiological principles are considered, it is not possible to establish a unique design and recommend a specific method of statistical analyses for the comparisons to be made. It is only possible to present guides that should be followed while performing the experiments. It is also recommended that a statistician be involved in the study.