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