PDA-确保微生物测试数据的完整性（一）
Microbiological Test Data – Assuring Data Integrity
微生物测试数据-确保数据的完整性
Edward Charles Tidswell and Tim Sandle
PDA Journal of Pharmaceutical Science and Technology 2017,
Access the most recent version at doi:10.5731/pdajpst.2017.008151
Microbiological Test Data – Assuring Data Integrity
微生物测试数据–确保数据的完整性
Abstract 摘要
Marketed drugs and devices possess specifications including critical microbiological quality attributes purposed to assure efficacy and patient safety.
已上市的药物和器械具有包括旨在确保疗效和患者安全的关键微生物质量属性在内的标准。
These attributes are legislated requirements intended to protect the recipient patient.
这些属性是立法规定的要求，旨在保护受体病人。
Sampling, microbiological testing, interpretation of data for final products, raw materials and intermediates all contribute to a cohesive assessment in the assurance of finished product quality.
抽样、微生物检测、最终产品、原材料和中间体数据的解释都有助于对最终产品质量的一致性评估。
Traditional culture-based microbiological methods possess inherent and unavoidable variability, recognized by the compendia and which might lead to erroneous conclusion pertaining to product quality.
传统的基于培养的微生物学方法具有固有的和不可避免的可变性，这一点已被药典所承认，并可能导致与产品质量有关的错误结论。
Such variability has been associated and intrinsically linked with data integrity issues; manufacturers have subsequently been encouraged by regulatory authorities to introduce multiple microbiologists or checks to prevent such issues.
这种可变性与数据完整性问题相关联，并在本质上联系在一起；随后，监管部门鼓励生产商引入多种微生物学家或检查方法，以防止此类问题的发生。
Understanding microbiological variability is essential such that genuine data integrity issues are identified.
了解微生物的可变性是至关重要的，这样才能确定真正的数据完整性问题。
Furthermore, a range of meaningful preventative strategies are feasible beyond increasing the capacity of the quality control microbiological laboratory.
此外，除了增加质量控制微生物实验室的能力之外，一系列有意义的预防策略是可行的。
This short review describes the legislative requirements, inherent microbiological variability and realistic actions and activities that genuinely assure patient safety.
本篇简短的综述描述了立法要求、内在的微生物多样性以及真正确保患者安全的现实行动和活动。
Key Words关键词
Microbiological tests, data integrity, microbiological quality attributes, sterility test, bioburden test
微生物测试，数据完整性，微生物质量属性，无菌测试，生物负载测试
Introduction 简介
The assessment of the microbiological quality of pharmaceutical finished drug products and devices is mandated through legislated requirements (21 CFR Parts 211 and 800 in the US and Volume 4 of Eudralex in the EU).
通过立法规定，对药品成品和器械的微生物质量进行评估。
These laws are purposed to help assure the safety of the recipient patient population from potential microbiological adulteration of drugs and devices.
这些法律的目的是帮助确保接受治疗的患者群体的安全，避免潜在的微生物掺假药物和器械。
Such adulteration may be regarded as adventitious contamination manifested as the non- sterility of items purported to be sterile or the possession of an item with a certain quantity of microorganisms (bioburden) or type of microorganism (objectionable) occurring within items marketed as non-sterile.
这种掺假可被视为不定之症污染，表现为声称无菌的物品不无菌，或拥有在作为非无菌商品销售的物品中含有一定数量的微生物(生物负载)或微生物类型(特定微生物)。
The actual technical, microbiological criteria for sterile and non-sterile products are regarded as microbiological quality attributes and defined in the pharmaceutical compendia recognized by the relevant Health Authority.
无菌和非无菌产品的实际技术、微生物学标准被视为微生物质量属性，并在相关卫生部门认可的药典中进行了定义。
It is against these standards by which legislated requirements governing a finished product or device are assessed and determined to be fit for market.
这是违反这些标准的立法要求，管理的成品或器械是评估和确定适合市场。
Similarly the pharmaceutical compendia detail the reference or referee method or analytical test by which a drug or device are sampled, tested and assessed against the microbiological criteria (or offer routes through which alternative methods can be adopted).
类似地，药典详细说明了参照或仲裁方法或分析测试，通过这种方法，药物或器械根据微生物学标准进行取样、测试和评估(或提供可采用其他方法的途径)。
Assurance of the microbiological quality of an item is not solely based upon end product sampling, testing and assessment; rather a complete cohesive and coordinated plan of process inputs (raw materials, additives, processing aids), process intermediates (including process steps such as intermediate hold) and environmental sampling, testing and assessment.
确保一项产品的微生物质量并不仅仅基于最终产品的抽样、测试和评估;而是一个完整的、连贯的、协调的过程投入(原材料、添加剂、加工助剂)、过程中间体(包括中间货舱等过程步骤)和环境取样、测试和评估的计划。
It is the complete, integrated and scientifically principled management of sampling, testing and assessment that provides the entire microbiological data which permits the disposition of product meeting its licensed label claim.
是对抽样、测试和评估的完整、综合和科学原则的管理，提供了完整的微生物数据，使产品符合其许可标签要求的处置。
This complete end-to-end cohort of data for each batch of finished drug product or device is also legislated through the cGMPs (for example 21 CFR Parts 211.110, and 211.113).
每批成品或器械的完整端到端队列数据也通过cGMPs进行立法(例如21 CFR第211.110和211.113部分)。
It is the combination of legislated requirements, established microbiological attributes,
sampling, testing and analysis which are purposed to help assure patient safety.
它是立法要求、已确立的微生物属性、为确保病人安全而进行的抽样、测试和分析。
Patient safety is the fundamentally crucial guiding principle and ethic in the provision of pharmaceutical finished drug products and devices.
在提供药物成品和器械方面，患者安全是根本的关键指导原则和伦理。
Industry and regulators are entirely aligned in this, and what could be more unethical and paradoxical than the provision of harmful therapies supplied to treat patients.
行业和监管机构在这一点上完全一致，而且可能比为病人提供有害疗法更不道德、更自相矛盾。
Equally the provision of safe therapies and assurance of patient safety in terms of microbiological quality is benefited with recognition of the following:
同样，由于认识到下列情况，为病人提供安全的治疗和保证病人的微生物质量也大有裨益:
Microbiological data does not per se materially control or modulate the bioburden or the
sterility of non-sterile products or products purported to be sterile.
微生物学数据本身并不能在实质上控制或调节生物群落非无菌产品或声称无菌的产品的无菌性。
Historically legislation has primarily relied upon data acquired from an analytical test as the proof of safety for the patient.
从历史上看，立法主要依靠从分析测试中获得的数据作为患者安全的证明。
This is understandable given the evidence-based foundation of judicial systems supporting legislation.
鉴于司法系统支持立法的证据基础，这是可以理解的。
In actuality the assurance of the microbiological quality of products is better served with an emphasis upon design and control (as set out in International Conference on Harmonization guidance Q8 Pharmaceutical development; Q9 Quality risk management; Q10 pharmaceutical quality system; Q11 Development and manufacture of drug substances).
事实上，设计和控制是保证产品微生物质量的关键（如ICHQ8药物开发；Q9质量风险管理；Q10药品质量体系；Q11药物的开发和制造）
Microbiological data has value; however this is only one part of the overall design, controls, checks and balances, necessary for assuring microbiological product quality.
微生物数据有价值;然而，这只是整体设计、控制、检查和平衡的一部分，是保证微生物产品质量所必需的。
Acquisition of microbiological data is reliant upon a sample upon which a particular test is
performed.
微生物数据的获取依赖于特定测试所依赖的样本执行。
Meaningful microbiological data providing supporting evidence that a well- designed process is in control and that the end product meets the required microbiological attribute can only be gained if a representative sample is obtained.
只有获得具有代表性的样品，才能获得有意义的微生物数据，这些数据提供了有力的证据，证明精心设计的过程处于控制之中，最终产品满足所需的微生物属性。
Samples must not only be authentic of the material tested, that is free from contamination during the activity of sampling but removed at a location and time that serves the purpose of the test (process control or material meeting a microbiological attribute).
样品不仅必须是所测试材料的真实样品，在取样过程中不受污染，而且必须在符合测试目的的地点和时间(过程控制或符合微生物特性的材料)被移除。
Typically a ‘worst case’ time for sampling is at the end of any process hold period.
通常，采样的“最坏情况”是在任何过程保持期的末尾。
Additionally the sample must be representative of material and account for any lack of homogeneity.
此外，样品必须具有代表性的材料和解释任何缺乏同质性。
An end-to-end integrated sampling plan based upon a genuine understanding of microbiological risk and control is the means to achieve this (Tidswell, 2017).
基于真正理解微生物风险和控制的端到端集成采样计划是实现这一目标的手段(Tidswell, 2017)。
Each sample is subject to a particular test methodology that is reliant upon a suitable method
development, qualification, and accompanying controls.
每个样本都要遵循特定的测试方法，该方法依赖于合适的方法开发、确认和伴随的控制。
Execution of the test is highly dependent upon the adequacy of the associated standard operating procedures (SOP) and the acumen of the microbiologist performing the method.
测试的执行高度依赖于相关标准操作规程(SOP)的充分性和微生物学家执行方法的敏锐性。
Finally once a sample has been tested the test is inspected and microbiological data is interpreted in order to make the microbiological assessment.
最后，一旦样品被测试，测试是检查和微生物数据的解释，以便作出微生物评估。
Traditional microbiological test methods invariably rely upon the visual inspection of a liquid or solid medium and either the enumeration of colony forming units (CFUs), an assessment of the presence/absence of CFUs or the presence of turbidity in liquid media.
传统的微生物测试方法总是依赖于对液体或固体培养基的目视检查，以及菌落形成单位(CFUs)的计数，对CFUs的存在/不存在或液体培养基中是否存在浑浊度的评估。
The CFU is an arbitrary estimate at best since the only cells able to form colonies are those that can grow under the conditions of the test.
CFU充其量是一个武断的估计，因为唯一能够形成菌落的细胞是那些在测试条件下能够生长的细胞。
These test conditions include: incubation temperature; incubation time; type of culture media; oxygen conditions and so on.
这些测试条件包括:培养温度;培养时间;培养基类型;氧气条件等等。
A subsequent recorded number of colony forming units (CFU) are not raw data but rather the
CFU is the microbiologist’s interpretation of the number of colonies on the plate (Sutton, 2012).
随后记录的菌落形成单元数(CFU)不是原始数据，而是CFU是微生物学家对平板上菌落数量的解释(Sutton, 2012)。
This interpretation is affected by the number of colonies present, where there is too high a number leads to counting errors as a result of confluence or overcrowding across the surface of the plate; and where the number is too low error arises because the CFU’s follow Poisson distribution.
这种解释受到当前菌落数量的影响，菌落数量过高会导致由于板块表面的融合或过度拥挤而导致计数错误;由于CFU遵循泊松分布，所以误差很小。
Of late several competent regulatory authorities have made inspectional observations of firms that are applying the microbiological test in alignment to the legislated cGMPs.
最近，几个主管管理当局对正在按照立法的cGMPs进行微生物检验的公司进行了检查。
Those observations note that a single microbiologist is visually inspecting the test and making an interpretation of the test.
这些观察结果表明，一个微生物学家正在视觉上检查测试并对测试进行解释。
For example, there have been regulatory comments where colony forming units have been miscounted.
例如，有监管部门的评论指出，菌落形成单位的数量被错误计算。
This can arise due to inappropriate light or magnification; a failure to use a suitable counting device; the incorrect multiplication of a dilution and so on.
这可能是由于不适当的光或放大；未能使用适当的计数装置;稀释倍数的错误乘法，等等。
These observations have been categorized as data integrity issues.
这些观察结果被归类为数据完整性问题。
It has thus become popular to suggest that deployment of multiple microbiologists is an effective and expedient preventative action of microbiological data integrity issues.
因此，建议部署多个微生物学家是微生物数据完整性问题的有效和方便的预防措施，这已成为流行的观点。
At first glance this may appear the most appropriate rectification; however this in itself is not a ubiquitous solution, may not be necessary and may in actuality prevent realization of our common goal of reduced patient risk, and maximized patient safety.
乍一看，这似乎是最恰当的纠正，然而，这本身并不是一个普遍存在的解决方案，可能没有必要，而且实际上可能会阻碍我们实现降低患者风险和最大化患者安全的共同目标。
Our necessary history of evidence-based judicial systems and legislation founded upon indisputable and uncontestable data (laws of evidence or the rule of evidence) has helped to serve our industry well, however it does not match well with microbiological data with its well- recognized inherent variability (Sutton, 2011a).我们建立在无可争议的数据(证据法或证据规则)基础上的以证据为基础的司法系统和立法的必要历史有助于我们的行业更好地服务，然而，由于其固有的可变性，它与微生物数据并不匹配(Sutton, 2011a)。
Variability can be divided in to two categories (Singer and Sutton, 2011):
可变性可分为两类(Singer和Sutton, 2011):
a) “Avoidable” variability (variability due to poor practice (Sutton 2010))
“可避免的”变异性(由于实践不当造成的变异性(Sutton 2010))
b) Inherently unavoidable variability (variability due to limitations of the methods and the vagaries of dealing with biological samples (Jarvis, 1989).
固有的不可避免的变异性(由于方法的限制和处理生物样本的反复无常而产生的变异性)(Jarvis, 1989)。
Assurance of microbiological quality is not primarily achieved through test data.
微生物质量的保证主要不是通过测试数据来实现的。
It frequently appears confoundlingly difficult to reconcile the constraints of microbiological tests, and interpretative assessment of those tests with the precise details of design control, and sampling.
要调和微生物试验的限制条件，以及对这些试验的解释性评估与设计控制和抽样的精确细节之间的矛盾，常常显得非常困难。
There is a need to ensure the data collected and purposed to demonstrate process control and product achieving microbiological quality attributes is as accurate as feasible, however there are many alternative means to achieve this other than multiple microbiologists performing the inspection and assessment.
有必要确保所收集和用于演示过程控制和产品实现微生物质量属性的数据尽可能准确可行，然而，除了多个微生物学家进行检查和评估之外，还有许多其他方法可以实现这一目标。
This short review is purposed to illustrate the inherent variability of culture based microbiological test methods (which cannot be reduced with any significance) and provide the range of options to ensuring the avoidance of potential data integrity issues.
这篇简短的综述旨在说明基于培养的微生物测试方法的固有可变性(不能有任何意义地降低这种可变性)，并提供了确保避免潜在数据完整性问题的选项范围。
Microbial Quality Attributes
微生物质量属性
All finished drug products and medical devices must be fit for use; granted marketing licenses based upon appropriate specifications, in combination with well-designed and controlled manufacturing processes conforming to the legislated cGMPs.
所有成品药品和医疗器械必须适合使用;根据适当的规范，结合符合法规cGMPs的精心设计和控制的生产过程，授予销售许可证。
For finished drug products this means a therapy must have appropriate characteristics of safety, identity, strength, purity, and quality.
对于成品药物，这意味着治疗必须具有适当的安全性、同一性、强度、纯度和质量特征。
Microorganisms (viable and non-viable), their products, or their inherent biochemical and physiological activity on or within therapies may adversely impact safety, strength, purity and quality.
：微生物(活的和不活的)、它们的产品，或它们在治疗上或治疗内固有的生化和生理活性可能对安全性、强度、纯度和质量产生不利影响。
Unlike an inanimate chemical or physical impurities or contaminants, the animate and dynamic nature of microorganisms posse a far greater risk to fitness for use and ultimately patient safety.
与无生命的化学或物理杂质或污染物不同，微生物的生命和动态特性对适宜使用和最终患者的安全构成了更大的风险。
In a certain quantity certain species of microorganisms may cause infection when introduced through the vehicle of a therapy, metabolically active microorganisms may transform a drug products formulation resulting in chemical changes, or changes in efficacy.
在一定数量的情况下，某些种类的微生物在通过治疗载体引入时可能引起感染，代谢活性微生物可以改变药物的配方，导致化学变化，或功效的变化。
Lastly the vestiges of microorganisms or sloughed off cell well and cell membrane may cause pyrogenic or toxic effect.
最后，微生物的残留或脱落的细胞和细胞膜良好，可能导致发热或毒性作用。
The microbial requirements of finished product specifications are therefore arguably the most important and are often underappreciated in their potential to adversely affect the recipient patient population.
因此，成品规范中的微生物要求可以说是最重要的，但由于其潜在的对受体患者群体产生不利影响的可能性，这些要求常常被低估。
Concomitantly pharmacopoeial compendia are very clear in stating the necessary microbial specifications of finished drug products.
与此同时，药典在说明成品的必要微生物规范方面也非常清楚。
Sterility is required for all parenteral products are stipulated in by the United States Pharmacopeia (USP), (<71> Sterility tests), Pharmacopoeia Europa (EP), (2.6.1 Sterility), and Japanese Pharmacopoeia (JP), (4.06 Sterility test).
美国药典(USP)、(<71>无菌试验)、欧洲药典(EP)、(2.6.1无菌)、日本药典(JP)、(4.06无菌试验)均规定注射用药物产品必须无菌。
Non-sterile finished drug product are permitted a level of bioburden which is based upon the dosage form and route of administration and stipulated in USP (<1111> Microbiological examination of non-sterile products: acceptance criteria for pharmaceutical preparations and substances), EP (2.6.12 Microbiological examination of non-sterile products: total; viable aerobic count) and JP (4.05 Microbial limit test).
在USP（<1111>），EP（2.6.12），JP（4.05）中，非无菌成品按规定的剂型和给药途径允许有一定的生物负载。
The potential impact of microorganisms on or within therapies is undisputable and clearly represents a source of risk to patient safety; however it is imperative that we recognize the precise specifications for bioburden as described by the industry standard pharmacopeia.
微生物对治疗或治疗内的潜在影响是毋庸置疑的，并清楚地代表了对患者安全的风险来源;然而，我们必须认识到的准确规范的生物负载所描述的行业标准药典。
USP <1111>, EP 2.6.12 EP and JP 4.05 list out the maximum limits of bioburden defined as total aerobic microbial counts (TAMC) and total yeast and mold count (TYMC) within different dosage forms of drug products; categorized as 10 CFU, 100CFUCFU and 1000CFU.
USP <1111>， EP 2.6.12 EP和JP 4.05列出了不同剂型药物中生物负载的最大限值，生物负载定义为总需氧微生物计数(TAMC)和总酵母和霉菌计数(TYMC);分为10 CFU, 100CFU和1000CFU。
Each pharmaceutical compendia subsequently qualifies the interpretation of these maximum bioburden limits; the three harmonized compendia (USP, EP, JP) all stating:
每个药典随后都有确认解释这些最大生物负载限度;三个协调案(USP, EP, JP)均声明:
“When an acceptance criterion for microbiological quality is prescribed, it is interpreted as follows:
“规定微生物质量验收标准时，解释为:
101cfu: maximum acceptable count = 20;
102cfu: maximum acceptable count = 200;
103 cfu: maximum acceptable count = 2000;
and so forth.”
All compendia provide due and appropriate recognition to the inherent variability of microbiology and microorganisms; the expectation is that the expert microbiologist appropriately interprets the data.
所有的药典都对微生物学和微生物的固有变异性提供了适当的认识;期望微生物学家专家能恰当地解释这些数据。
A 200% leeway is permissible which is essentially the acceptance of a two-fold difference or variability in the enumeration of CFUs.
允许有200%的偏差，这本质上是接受CFUs计数中的双重差异或变异性。
This variability is necessary because of the inherent dynamic nature of microorganisms, microbial populations and the means by which samples are handled and tested by culture-based methods.
这种变化是必要的，因为微生物的固有动态性质、微生物种群以及用培养方法处理和测试样品的方法。
The same is true for process inputs (raw materials, additives, processing aids), process intermediates (including process steps such as intermediate hold) and environmental testing; enumeration to established microbial quality attributes is an important component of quality assurance however there is recognized variability.
工艺输入(原料、添加剂、助剂)、工艺中间体(包括中间货舱等工艺步骤)和环境试验也是如此;对已建立的微生物质量属性的枚举是质量保证的一个重要组成部分，但也存在公认的可变性。
In addition to microbial quality attributes the pharmacopoeial compendia detail the standardized methodology of testing a sample purposed to enumerate bioburden or establish a presence or absence.
除了微生物质量属性外，药典还详细说明了用于列举生物样本或确定存在或不存在的样本检测的标准化方法。
These methods are recognized as the referee or reference method which a firm needs to optimize and qualify for each of their sample formulations assuring the recovery of any resident culturable (using the media and conditions stated) microorganisms; the microbiology quality control (QC) laboratory tests.
这些方法被认为是裁判或参考方法，公司需要对其每一种样品配方进行优化和确认，以确保回收任何可培养的(使用所述的培养基和条件)微生物;微生物学质量控制(QC)实验室检测。
（未完待续）
下节预告：PDA-确保微生物测试数据完整性（二）
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