Real-time PCR-based method for measuring the phase variation frequency of flagellin in Salmonella enterica serovar Typhimurium

IF 2.5 4区生物学 Q4 BIOCHEMICAL RESEARCH METHODS

Journal of microbiological methods Pub Date : 2026-02-01 Epub Date: 2025-12-13 DOI:10.1016/j.mimet.2025.107362

Momoko Nakayama , Nobuo Arai , Yohsuke Ogawa , Masahiro Kusumoto , Masahiro Eguchi

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Abstract

Some Salmonella serovars possess two antigenic phases of the H antigen, known as phase 1 and phase 2. These phases reversibly switch between each other at a constant frequency; phase variation frequency. Previously, we focused on the inversion of hin and developed a real-time PCR system, H-qPCR, to quantitatively measure the proportion of bacterial cells expressing each phase of flagellin for Salmonella enterica serovar Typhimurium (S. Typhimurium). In this study, we developed a novel method to analyze the phase variation frequency of H antigen of S. Typhimurium, based on the H-qPCR approach. It was shown that our method offered equivalent accuracy but superior sensitivity compared with the conventional method for measuring the phase variation frequency. Our real-time PCR-based method is expected to contribute to the analysis of phase variation frequency as a useful tool.

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基于实时荧光定量pcr的肠炎沙门氏菌血清鼠伤寒沙门氏菌中鞭毛蛋白相位变化频率测定方法。

一些沙门氏菌血清型具有H抗原的两个抗原阶段，称为第1期和第2期。这些相位以恒定的频率在彼此之间可逆地切换；相位变化频率。在此之前，我们专注于hin的倒置，并开发了一种实时PCR系统，H-qPCR，用于定量测量肠沙门氏菌血清鼠伤寒沙门氏菌（S. Typhimurium）中表达鞭毛蛋白各相的细菌细胞比例。本研究建立了一种基于H- qpcr的鼠伤寒沙门氏菌H抗原相位变化频率分析方法。结果表明，与传统的相位变化频率测量方法相比，该方法具有相当的精度和较高的灵敏度。我们基于实时pcr的方法有望作为一种有用的工具，为相位变化频率的分析做出贡献。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Journal of microbiological methods 生物-生化研究方法

CiteScore

4.30

自引率

4.50%

发文量

151

审稿时长

29 days

期刊介绍： The Journal of Microbiological Methods publishes scholarly and original articles, notes and review articles. These articles must include novel and/or state-of-the-art methods, or significant improvements to existing methods. Novel and innovative applications of current methods that are validated and useful will also be published. JMM strives for scholarship, innovation and excellence. This demands scientific rigour, the best available methods and technologies, correctly replicated experiments/tests, the inclusion of proper controls, calibrations, and the correct statistical analysis. The presentation of the data must support the interpretation of the method/approach. All aspects of microbiology are covered, except virology. These include agricultural microbiology, applied and environmental microbiology, bioassays, bioinformatics, biotechnology, biochemical microbiology, clinical microbiology, diagnostics, food monitoring and quality control microbiology, microbial genetics and genomics, geomicrobiology, microbiome methods regardless of habitat, high through-put sequencing methods and analysis, microbial pathogenesis and host responses, metabolomics, metagenomics, metaproteomics, microbial ecology and diversity, microbial physiology, microbial ultra-structure, microscopic and imaging methods, molecular microbiology, mycology, novel mathematical microbiology and modelling, parasitology, plant-microbe interactions, protein markers/profiles, proteomics, pyrosequencing, public health microbiology, radioisotopes applied to microbiology, robotics applied to microbiological methods,rumen microbiology, microbiological methods for space missions and extreme environments, sampling methods and samplers, soil and sediment microbiology, transcriptomics, veterinary microbiology, sero-diagnostics and typing/identification.