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[一种用于评估表面消毒程序中检测病原体回收的简单冲洗技术]

[A simple wash technic for the recovery of test pathogens in the evaluation of surface disinfection procedures].

作者信息

Koller W

出版信息

Zentralbl Bakteriol Mikrobiol Hyg B. 1984 May;179(2):112-24.

PMID:6377750
Abstract

Methods currently recommended for quantitative bacteriological sampling of surfaces usually suffer from either unreliable results (plain swabbing techniques and contact cultures) or high expenses in terms of material, equipment and/or labour (rinse techniques using sampling fluids). Therefore, a modified swab-rinse technique (SRT) was devised as a reasonably precise and simple alternative: A small amount of sampling fluid (1.5 ml), which can contain neutralisers, is transferred onto the flat surface under investigation; with this fluid and a pre-moistened small cotton-swab an area of 3 cm diameter is thoroughly washed for 15 s. Then 0.1 ml- and 0.5 ml-amounts of the washliquid are collected with automatic microliter pipettes and transferred and spread onto Casein-Soy-Agar for enumeration. In parallel experiments with contact cultures (rodac plates), the new SRT up to 3000 cfu per sample exhibited a linear answer to increasing inocula of E. coli on tiles (Fig. 1, rings). Rodac-plates proved to possess a rather limited span of reliable counts: above 100 colonies, increasing numbers of bacteria were prevented from forming distinguishable colonies (Fig. 1, dots and +es); thus, colony counts higher than 100 on rodac plates cannot be expected to be true estimates. In addition, in the higher count range the results of a common, simplified counting technique differed markedly from results of true counts (Fig. 2). Both methods were equally sensitive in detecting low counts of bacteria (Fig. 1, Fig. 3). Shake-rinse techniques using high volumes of sampling fluid provide lower sensitivity than contact cultures or the new SRT: The use of 100 ml of sampling fluid (15) and plating of 0.1 ml- or 1.0 ml-aliquots of sampling fluid keeps the threshold for detection of bacteria as high as 1000 and 100 per sample, respectively (Fig.3); nevertheless, sensitivity of shake-rinse techniques can be increased by filtration of the whole sample. Thus, the new swab-rinse technique combines several advantages: wide span of true estimates since washliquid can be diluted for enumeration of high counts; high sensitivity ( = ability to detect small numbers of testbacteria in sample) since about one half of the sample is plated; good recovery of testbacteria from both smooth and coarse surfaces; simplicity; the new swab-rinse technique requires basic laboratory equipment and ordinary media and no shaking- or filtration devices; option for immediate and strong neutralisation of disinfectant residues; the sampling fluid can contain any neutraliser; option for automated colony counts since any kind of petri-dishes can be used for culture.

摘要

目前推荐用于表面定量细菌学采样的方法通常存在以下问题

要么结果不可靠(普通擦拭技术和接触培养法),要么在材料、设备和/或人力方面成本高昂(使用采样液的冲洗技术)。因此,设计了一种改良的拭子 - 冲洗技术(SRT)作为一种合理精确且简单的替代方法:将少量(1.5毫升)可含有中和剂的采样液转移到被研究的平面上;用这种液体和预先湿润的小棉签对直径3厘米的区域彻底冲洗15秒。然后用自动微量移液器收集0.1毫升和0.5毫升的冲洗液,并转移到酪蛋白大豆琼脂上进行计数。在与接触培养法(罗达克平板)的平行实验中,新的SRT在每个样本高达3000 cfu的情况下,对瓷砖上大肠杆菌接种量的增加呈现出线性响应(图1,环形)。罗达克平板被证明可靠计数的范围相当有限:超过100个菌落时,越来越多的细菌无法形成可区分的菌落(图1,点和 + 号);因此,罗达克平板上高于100的菌落计数不能被视为真实估计值。此外,在较高计数范围内,一种常见的简化计数技术的结果与真实计数结果明显不同(图2)。两种方法在检测低细菌计数时同样敏感(图1,图3)。使用大量采样液的振荡冲洗技术比接触培养法或新的SRT灵敏度低:使用100毫升采样液(15)并接种0.1毫升或1.0毫升采样液等分试样,检测细菌的阈值分别高达每个样本1000和100(图3);然而,振荡冲洗技术的灵敏度可通过对整个样本进行过滤来提高。因此,新的拭子 - 冲洗技术具有多个优点:由于冲洗液可稀释用于高计数的计数,真实估计范围广;灵敏度高(=能够检测样本中的少量测试细菌),因为大约一半的样本用于接种;能从光滑和粗糙表面良好地回收测试细菌;操作简单;新的拭子 - 冲洗技术需要基本的实验室设备和普通培养基,无需振荡或过滤装置;可立即强烈中和消毒剂残留;采样液可包含任何中和剂;可进行自动菌落计数,因为任何类型的培养皿都可用于培养。

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