使用核酸模拟探针的流式荧光原位杂交技术检测细菌。
Flow-FISH Using Nucleic Acid Mimic Probes for the Detection of Bacteria.
机构信息
LEPABE - Laboratory for Process Engineering, Environment, Biotechnology and Energy, Department of Chemical Engineering, Faculty of Engineering, University of Porto, Porto, Portugal.
i3S - Instituto de Investigação e Inovação em Saúde, Universidade do Porto, Porto, Portugal.
出版信息
Methods Mol Biol. 2021;2246:263-277. doi: 10.1007/978-1-0716-1115-9_17.
Flow-Fluorescence in situ hybridization (Flow-FISH) enables multiparametric high-throughput detection of target nucleic acid sequences at the single cell-level, allowing an accurate quantification of different cell populations by using a combination of flow cytometry and fluorescent in situ hybridization (FISH). In this chapter, a flow-FISH protocol is described with labeled nucleic acid mimics (NAMs) (e.g. LNA/2'OMe and PNA) acting as the reporter molecules. This protocol allows for the specific detection of bacterial cells. Hence, this protocol can be carried out with minor adjustments, in order to simultaneously detect different species of bacteria in different types of clinical, food, or environmental samples.
流式荧光原位杂交（Flow-FISH）使我们能够在单细胞水平上高通量地检测目标核酸序列，通过流式细胞术和荧光原位杂交（FISH）的结合，实现对不同细胞群的准确定量。在本章中，我们描述了一种使用标记核酸模拟物（NAMs）（如 LNA/2'OMe 和 PNA）作为报告分子的 Flow-FISH 方案。该方案允许对细菌细胞进行特异性检测。因此，该方案可以进行微小调整，以便同时检测不同类型的临床、食品或环境样本中的不同细菌种类。
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