利用肽核酸的荧光原位杂交检测微生物。

Detection of Microorganisms by Fluorescence In Situ Hybridization Using Peptide Nucleic Acid.

机构信息

LEPABE - Laboratory for Process Engineering, Environment, Biotechnology and Energy, Faculty of Engineering, University of Porto, Porto, Portugal.

INIAV - National Institute for Agrarian and Veterinarian Research, Vairao, Portugal.

出版信息

Methods Mol Biol. 2020;2105:217-230. doi: 10.1007/978-1-0716-0243-0_13.

DOI:10.1007/978-1-0716-0243-0_13

PMID:32088873

Abstract

Fluorescence in situ hybridization (FISH) is a 30-year-old technology that has evolved continuously and is now one of the most well-established molecular biology techniques. Traditionally, DNA probes are used for in situ hybridization. However, synthetic molecules are emerging as very promising alternatives, providing better hybridization performance and making FISH procedures easier and more efficient. In this chapter, we describe a universal FISH protocol, using nucleic acid probes, for the detection of bacteria. This protocol should be easily applied to different microorganisms as a way of identifying in situ relevant microorganisms (including pathogens) and their distribution patterns in different types of samples.

摘要

荧光原位杂交（FISH）是一项拥有 30 年历史的技术，它不断发展，现已成为最成熟的分子生物学技术之一。传统上，DNA 探针用于原位杂交。然而，合成分子作为非常有前途的替代品正在出现，它们提供了更好的杂交性能，使 FISH 程序更简单、更高效。在本章中，我们描述了一种使用核酸探针的通用 FISH 方案，用于检测细菌。该方案应易于应用于不同的微生物，作为一种识别原位相关微生物（包括病原体）及其在不同类型样本中分布模式的方法。