Rapid and cost-effective method for microorganism enumeration based on miniaturization of the conventional plate-counting technique
Rapid and cost-effective method for microorganism enumeration based on miniaturization of the conventional plate-counting technique
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DOI：
10.1051/lait:2006005
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发表时间：
2006-05-01
期刊：
影响因子：
--
通讯作者：
Gautier, M
中科院分区：
文献类型：
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作者：
Baron, F;Cochet, MF;Gautier, M
传统的平板计数技术虽然仍然在需要微生物筛查或计数的实验室中常规使用，但在分析大量样品时对于操作员来说仍然耗时且令人厌烦。我们开发了一种小型化方法，该方法基于在 96 孔微孔板中连续十倍稀释样品，然后通过琼脂覆盖程序多次倒入 24 孔微孔板中，从而提供轻松的菌落计数和解释。使用传统的平板计数技术作为参考，对亚致死热应激下的大肠杆菌细胞活力测量、牛奶发酵过程中的乳酸菌计数以及变质食品中的总需氧菌群、大肠菌群和金黄色葡萄球菌含量进行分析，测试了这种小型化方法的准确性、灵敏度和重现性。无论使用何种细菌和培养基，用我们的小型化方法和传统方法获得的结果都没有发现显着差异。这种可靠的小型化方法可以降低总体成本和处理时间，对于涉及大范围细胞浓度以及大量样品和重复的实验很有用。
The conventional plate-counting technique, while still routinely used in laboratories requiring micro-organism screening or enumeration, remains time-consuming and tiresome for the operator when large numbers of samples are assayed. We developed a miniaturized method based on serial tenfold dilution of samples in 96-well micro-plates followed by multiple pour-plating into 24-well micro-plates with an agar overlay procedure, providing easy colony counting and interpretation. The accuracy, sensitivity and reproducibility of this miniaturized method were tested using the conventional plate-counting technique as reference for viability measurements of Escherichia coli cells submitted to a sub-lethal thermic stress, enumeration of lactic bacteria during milk fermentation, and analysis of the total aerobic flora, coliform and Staphylococcus aureus content of spoiled food. No significant differences were found between the results obtained with our miniaturized method and the conventional one, regardless of the bacteria and media used. Providing a reduction in overall cost and handling time, this reliable miniaturized method should be useful for experiments involving large ranges of cell concentration as well as great numbers of samples and replicates.