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用于分子生物学分析的水样三种浓缩方案的回收率比较:膜过滤、纱布垫过滤和离心。

Comparison between the recovery rate of three concentration protocols of water samples intended for analysis by Molecular Biology: Membrane filtration, filtration on gauze pad and centrifugation.

作者信息

El Boujnouni Hamoucha, Nait Balla Kaoutar, Belkadi Bouchra, Rahouti Mohamed

机构信息

Team of Botany and Valorization of Plant and Fungal Resources Plant and Microbial Biotechnologies, Biodiversity and Environment Research Center, Faculty of Sciences, Mohammed V University in Rabat, B.P. 1014 RP, Rabat, Morocco.

Team of Microbiology and Molecular Biology, Plant and Microbial Biotechnologies, Biodiversity and Environment Research Center, Faculty of Sciences, Mohammed V University in Rabat, B.P. 1014 RP, Rabat, Morocco.

出版信息

Saudi J Biol Sci. 2022 Mar;29(3):1592-1597. doi: 10.1016/j.sjbs.2021.11.004. Epub 2021 Nov 11.

Abstract

The presence of microorganisms (bacteria, protozoa, viruses, etc.) in water is a crucial indicator of its quality and safety. The detection of these microorganisms by conventional and classical techniques is widely used in water quality control laboratories; nevertheless these methods have limitations in terms of rapidity and precision of results. The use of Molecular Biology has been a great evolution in the techniques of water analysis. However, the choice of the concentration protocol allowing for the best rate of microorganism recovery in a suspension remains a real challenge. The objective of this experimental study is to compare the recovery rate of three different protocols of water concentration (membrane filtration, filtration on gauze pad and centrifugation) for samples intended for analysis by polymerase chain reaction PCR. Which can then serve as a reference protocol for water quality control laboratories. The experimental results have shown that the membrane filtration protocol yields the best recovery rate and concentration of microorganisms followed by filtration on gauze pad, while the centrifugation protocol (8000, 10 min, 22 °C) gives the lowest rate of recovery out of the three protocols. The experimental results obtained through this study allows us to contribute to the optimization and standardization of water samples concentration techniques intended for analysis by Molecular Biology.

摘要

水中微生物(细菌、原生动物、病毒等)的存在是其质量和安全性的关键指标。传统和经典技术对这些微生物的检测在水质控制实验室中广泛使用;然而,这些方法在结果的快速性和精确性方面存在局限性。分子生物学的应用是水分析技术的一大进步。然而,选择能在悬浮液中实现最佳微生物回收率的浓缩方案仍然是一个实际挑战。本实验研究的目的是比较三种不同的水样浓缩方案(膜过滤、纱布垫过滤和离心)对用于聚合酶链反应(PCR)分析的样品的回收率。进而可为水质控制实验室提供一个参考方案。实验结果表明,膜过滤方案产生的微生物回收率和浓度最佳,其次是纱布垫过滤,而离心方案(8000转/分钟,10分钟,22°C)在这三种方案中回收率最低。通过本研究获得的实验结果有助于优化和标准化用于分子生物学分析的水样浓缩技术。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c233/8913413/7e885efd2c4c/gr1.jpg

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