Quantitative method for measuring the proportion of bacterial cells expressing phase 1 or 2 of flagellin of Salmonella enterica serovar Typhimurium

IF 1.7 4区生物学 Q4 BIOCHEMICAL RESEARCH METHODS

Journal of microbiological methods Pub Date : 2024-08-10 DOI:10.1016/j.mimet.2024.107013

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

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引用次数: 0

Abstract

Salmonella enterica subsp. enterica is a major pathogen that causes zoonotic foodborne diseases worldwide. Some Salmonella serovars possess two antigenic phases for flagellin: phase 1 and 2. In Salmonella enterica serovar Typhimurium (S. Typhimurium), the flagellin is antigenically divided into “Hi” as phase 1 and “H1 or H2” as phase 2. Flagellin phase variation is regulated by inversion of hin gene. We focused on the inversion of hin and developed a real-time PCR system to quantitatively measure the proportion of bacterial cells expressing each phase of flagellin. In this study, we demonstrated that our newly developed real-time PCR system shows high quantitative accuracy and aligns with flagellin expression status. Furthermore, the newly developed real-time PCR system was applicable to various S. Typhimurium laboratory and field strains. This newly developed real-time PCR system has the potential to become a powerful tool for analyzing flagellin phase variation.

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定量测量表达鼠伤寒沙门氏菌鞭毛蛋白 1 期或 2 期的细菌细胞比例的方法。

肠炎沙门氏菌亚种是导致全球人畜共患食源性疾病的主要病原体。一些沙门氏菌血清型具有两个鞭毛蛋白抗原阶段：第 1 阶段和第 2 阶段。在鼠伤寒沙门氏菌（S. typhimurium）中，鞭毛蛋白抗原分为 "Hi"（1 期）和 "H1 或 H2"（2 期）。鞭毛蛋白的相位变化受 hin 基因反转的调控。我们重点研究了 hin 基因的反转，并开发了一种实时 PCR 系统来定量测量表达各相鞭毛蛋白的细菌细胞比例。在这项研究中，我们证明了新开发的实时 PCR 系统具有很高的定量准确性，并且与鞭毛蛋白的表达状态一致。此外，新开发的实时 PCR 系统适用于各种鼠伤寒杆菌实验室菌株和野外菌株。这一新开发的实时 PCR 系统有望成为分析鞭毛蛋白阶段变异的有力工具。

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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.