Identification and Physiological and Biochemical Characteristics Analysis of A Foodborne Pathogenic Bacterium by 16S rRNA Combined with Tuf and RapC Gene Sequencing and Evolutionary Tree Construction
10.13241/j.cnki.pmb.2025.20.002
- VernacularTitle:16S rRNA结合tuf和rapC基因测序及进化树构建对一株食源性致病菌的鉴定及生理生化特性分析
- Author:
Yan-rong GUO
1
;
Li LI
;
Lei CHEN
Author Information
1. 上海震旦职业学院公共卫生与护理学院 上海 201908
- Publication Type:Journal Article
- Keywords:
Bacillus;
16S small subunit ribosomal ribonucleic acid;
Evolutionary tree;
Gene sequencing;
Strain identification;
Physiological and biochemical characteristics
- From:
Progress in Modern Biomedicine
2025;25(20):3211-3219
- CountryChina
- Language:Chinese
-
Abstract:
Objective:This study identified a foodborne pathogen Foodborne Pathogenic Bacterium by 16S ribosomal ribonucleic acid(16S rRNA)combined with TU translation elongation factor(tuf)and recombinantallophycocyanin(rapC)gene sequencing and constructing an evolutionary tree,and analyze its physiological and biochemical characteristics.Methods:A anal swab sample was collected from an adult patient with vomiting and diarrhea admitted to Ruijin Hospital affiliated with Shanghai Jiao Tong University School of Medicine.Separated and cultivated single colonies as test strains,Observe its morphological changes and analyze it was physiological and biochemical characteristics.Using 16S rRNA,tuf and rapC gene sequencing,and constructing an evolutionary tree.Identification of bacterial strains was used MADLI-TOF-MS mass spectrometer.Result:A Gram positive bacterium was isolated from an adult anal swab with symptoms of vomiting and diarrhea.The morphological observation results show that,Gram positive bacteria grow better in aerobic environment on blood agar plates,and grow slowly in anaerobic environment.The analysis result of physiological and biochemical characteristics shows that,strain SH1 was a facultative anaerobic bacterium that grows better under aerobic conditions to produce Bacillus subtilis.It was salt tolerant and heat-resistant,capable of oxidizing and fermenting various sugars to produce acid,and has the characteristics of utilizing citrate salts.The results of constructing an evolutionary tree of SH1 16S sequencing and related closely related strains showed that,the evolutionary tree can determine that SH1 belongs to the genus Bacillus,but it was specific species cannot be determined.The results of constructing an evolutionary tree of SH1 tuf sequencing and related closely related strains showed that,it was determined to had the closest relationship with Bacillus halotolerans strain F41-3,followed by Bacillus halotolerans strain KKD1 and Bacillus halotolerans strain XH-1.The results of constructing an evolutionary tree of SH1 rapC sequencing and related closely related strains showed that,it was determined to had the closest relationship with Bacillus halotolerans strain KKD1,followed by Bacillus halotolerans strain XH-1.The identification of mass spectrometry results showed that,the identified strains were Bacillus subtilis,Bacillus amyloliquefaciens,or Bacillus Vallrsrnortis.Conclusion:This study used 16S rRNA gene combined with tuf and rapC gene sequencing to constructing an evolutionary tree for rapid identification of a pathogenic bacterium is Bacillus halotolerans,to lay the foundation for subsequent studies on its toxicity and pathogenicity.
