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Establishment and evaluation of a method for efficient screening of Clostridium butyricum
Z. Yi, FA. Sadiq, H. Wang, J. Zhao, H. Zhang, W. Lu, W. Chen
Language English Country United States
Document type Journal Article
Grant support
JUSRP51903B
Fundamental Research Funds for the Central Universities
JUFSTR20180102
National first-class discipline program of Food Science and Technology
2019YFF0217601
National Key Research and Development Program of China
- MeSH
- Genes, Bacterial MeSH
- Bacteriological Techniques methods standards MeSH
- Clostridium butyricum genetics isolation & purification MeSH
- DNA Primers MeSH
- Species Specificity MeSH
- Feces microbiology MeSH
- Limit of Detection MeSH
- Polymerase Chain Reaction MeSH
- Publication type
- Journal Article MeSH
At present, the traditional methods for the screening of Clostridium butyricum are not sufficiently selective and efficient. Therefore, it is necessary to establish a targeted and efficient screening method for the detection of C. butyricum. Bioinformatics was used in this study to find C. butyricum specific genes, and species-specific primers were designed based on the conserved regions of the targeted genes, followed by optimization of the PCR conditions. Methodological evaluation was carried out, and the results were compared with the traditional screening method based on Trypticase Sulfite Neomycin (TSN) selective medium. A high-efficiency PCR screening method, targeting C. butyricum species-specific primers, was established. The method was confirmed to have high specificity and sensitivity towards C. butyricum cut-off CFU 103. Compared with the traditional method, the screening success rate of C. butyricum strains increased from 0.61 to 81.91%. The PCR screening method could quickly and accurately detect C. butyricum in samples and dramatically improve screening efficiency.
References provided by Crossref.org
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- $a At present, the traditional methods for the screening of Clostridium butyricum are not sufficiently selective and efficient. Therefore, it is necessary to establish a targeted and efficient screening method for the detection of C. butyricum. Bioinformatics was used in this study to find C. butyricum specific genes, and species-specific primers were designed based on the conserved regions of the targeted genes, followed by optimization of the PCR conditions. Methodological evaluation was carried out, and the results were compared with the traditional screening method based on Trypticase Sulfite Neomycin (TSN) selective medium. A high-efficiency PCR screening method, targeting C. butyricum species-specific primers, was established. The method was confirmed to have high specificity and sensitivity towards C. butyricum cut-off CFU 103. Compared with the traditional method, the screening success rate of C. butyricum strains increased from 0.61 to 81.91%. The PCR screening method could quickly and accurately detect C. butyricum in samples and dramatically improve screening efficiency.
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