生物负荷回收验证-生物负荷鉴定/筛选方法Ⅰ

Bioburden Characterization/Screening Methods
生物负荷鉴定/筛选方法

The level to which a manufacturer needs to characterize the bioburden population of a medical product or component is very closely related to the manufacturing process, sterilization process, and the end use of the product.
生产者需要表示中产品的生物负荷种群或成分的水平，这个水平与生产过程、灭菌过程以及产品的使用密切相关。

In general, the bioburden of a medical product should be sufficiently characterized to allow the manufacturer to analyze trends in the population level as well as the types of microorganisms which can be critical to the process. These organisms are of those varieties which produce spores or endotoxins or may multiply during the manufacturing process and can alter the product. For filter-sterilized products, the manufacturer must be concerned about controlling Gram negative bacteria, particularly Pseudomonas species, and the population level so that it does not exceed the capacity of the filter.
通常，医药产品的生物负荷应该允许生产者分析种群水平的趋势，和对过程至关重要的微生物类型。这些微生物是指那些能产生孢子、内毒素或能在生产期间繁殖并改变产品的种类。对于无菌过滤产品，生产者必须关注控制革兰氏阴性菌，尤其是假单胞菌，控制种群水平以便不会超过过滤能力。

The manufacturer may approach bioburden characterization in many different ways. The usual method, as discussed above, is through bioburden screening for microorganism population levels. Other methods include microorganism identification and testing for resistance to the sterilization process. A screening test may be more appropriate for certain items followed by more substantive testing based on the screen results. 
生产者可以用许多不同的方法处理生物负荷特征。如上所述的常用方法是通过生物负荷筛选微生物种群水平。其他方法包括对耐灭菌的微生物鉴定和测试。筛选检测可能更合适对某些种类，紧随其后的是更多以筛选结果为基础的实质性测试。

It may also be necessary to determine if the product or stages of the process promote changes in the population level. This requires knowledge of stages in the process where bioburden may be affected. During the testing for microbial levels, additional samples can be prepared from the dilutions or rinses for transfer to differential or selective media. It is also often practical to select isolates from the general purpose medium after enumeration for further characterization by staining or transfer to differential selective media.
决定是否产品或加工阶段能促进种群水平变化是必要的。这要求了解影响生物负荷的加工阶段。在测试微生物水平期间，从稀释液或者冲洗液中准备额外的样本，转移到不同的培养基或选择性培养基中。这对于从常用的目的培养基中挑选的分离菌，再通过染色和转入不同的选择培养基中进一步特征化，也是实用的。

Use of media selective for groups of Gram negative bacteria or Gram staining of a representative population can provide information on the potential for endotoxin presence. If significant numbers of Gram negative bacteria are present, various stages of the process/product should be evaluated for the presence of endotoxin and potential for microorganism proliferation.
对革兰氏阴性菌菌群或典型的种群的革兰氏染色使用选择培养基，能对内毒素出现的潜在性提供信息。如果出现大量革兰氏阴性菌，那么应该评估生产/加工的不同阶段中内毒素的出现和微生物增生的潜在性。

To detect anaerobic organisms additional samples may be incubated in an anaerobic environment. Colonies from these plates, however, should be evaluated through appropriate testing to determine if they are indeed strict anaerobes which have been recovered only from these incubation conditions, or if they are facultative organisms which have already been isolated during incubation in aerobic conditions.
为了检测厌氧微生物，在厌氧环境中培养附加样本。然而，这些平板中的克隆应该通过合适的检测方法评估，以决定它们是否是仅从这些培养条件下复原的严格厌氧微生物，或者它们是不是在需氧条件培养期间已经被分离的兼性微生物。

A procedure to detect and enumerate fungi separately may be desired. In this case, additional samples may be tested from each dilution using a growth medium specifically designed to support fungal growth. An example of this may be Soybean-Casein Digest agar, Sabouraud dextrose agar, or some other similar medium.
需要一个操作程序检测并列举真菌。在这种情况下，检测从每个稀释液中得到的附加样本，这些稀释液是使用了专门用于真菌生长的培养基。大豆消化酪素琼脂培养基、沙保氏葡萄糖琼脂、或者一些其他的类似培养基。

Incubation at elevated temperatures, such as 50-55℃ , may be used for the
enumeration of thermophiles. Other temperature ranges may be considered for other types of microorganisms.
嗜热菌需提高温度培养，例如50-55℃。对其他类型的微生物考虑用其他温度范围。

Incubation for extended times and at various temperatures should be done initially to determine if there are significant differences in recovery. This is particularly of concern where damaged cells may be present or if processing conditions may promote growth. A qualitative evaluation of the bioburden of a product or component to be sterilized can be accomplished by screening a sample of the item(s) to be sterilized for the resistance of their resident bioburden to the sterilization method. 
为了确定在复原中是否存在明显区别，开始就应该用不同的温度培养并延长培养时间。尤其应该注意损坏的细胞可能出现在哪或者是否处理条件能够促进生长。对一个已消毒的产品或者成分的生物负荷进行定性评估，是通过筛选一系列已消毒样本完成的，筛选这些已消毒样本的生物负荷对消毒方法的忍受力。

The screen is a shortened exposure to the sterilization cycle designed to eliminate those organisms in the indigenous bioburden which have no resistance to the sterilization and leave behind those organisms which have the potential to survive. The samples should be taken at apoi nt in the process immediately prior to the sterilization after completion of all manufacturing steps. 
对于消毒周期来说，筛选是缩短的暴露，所设计的消毒周期是为了去除那些固有生物负荷对消毒没有抵抗力的微生物，同时留下那些能够存活的微生物。应该在完成所有制造步骤以后，进行消毒步骤之前，在过程中的一个点取出样本。

Following exposure of the items to the abbreviated sterilization cycle, a recovery test in an appropriate medium (such as nutrient broth) is performed on a representative quantity of the sample(s). Any organisms growing out of the resistance screening are then tested for level of resistance to the sterilization treatment (i.e., D-value determination). 
简化的消毒周期之后，就可以在适当培养基（例如肉汤培养基）中进行大量的典型样本的复原测试。测定任何经抗性筛选生长的微生物的抗消毒水平（例如：D值法测定）。

The resistance of the isolates are then compared to the resistance of the biological indicator organisms used to validate the full sterilization cycle. Any organism found in the bioburden which is more resistant than the validation biological indicator represents a potential challenge to the process. At this point, quantitative analysis can be undertaken to evaluate whether the resistant organism is found in significant numbers.
隔离群的抗性与用于验证整个消毒周期的生物指示微生物对比。在生物负荷中发现的任何比验证指示生物更具抵抗力的微生物都对过程存在潜在挑战。根据这一点，进行定量分析以评估是否存在大量的抗性微生物。

       文章来源：药品微生物检测

       本网站刊载的所有内容，包括文字、图片、音频、视频、软件等，如非标注为“原创”，则相关版权归原作者所有，如原作者不愿意在本网站刊登相关内容，请及时通知本站，我们将第一时间予以删除。

曜分享

分享精彩资讯