Rapid Detection of Serratia fonticola by TaqMan Quantitative Real-Time PCR Using Primers Targeting the gyrB Gene
| Autor: | Ti yin Zhang, Zheng Teng, Jing Hua Ruan, Wu Jun Wang, Quan yang Bai, Zhi deng Zhang, Daojin Yu, Li Yun Wu, Huang Yifan |
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| Rok vydání: | 2017 |
| Předmět: |
0301 basic medicine
DNA Bacterial Serratia 030106 microbiology Real-Time Polymerase Chain Reaction Applied Microbiology and Biotechnology Microbiology Sensitivity and Specificity law.invention 03 medical and health sciences Bacterial Proteins law TaqMan Animals Humans Gene Polymerase chain reaction DNA Primers Genetics biology Nucleic acid sequence Reproducibility of Results General Medicine biology.organism_classification 16S ribosomal RNA Enterobacteriaceae Molecular biology 030104 developmental biology DNA Gyrase bacteria Primer (molecular biology) |
| Zdroj: | Current microbiology. 74(11) |
| ISSN: | 1432-0991 |
| Popis: | A gyrB gene is present in the majority of bacterial species, and encodes the ATPase domain of DNA gyraseB-subunit protein, which is essential for transcription and replication of bacteria. The gyrB gene exhibits higher nucleotide sequence variability than the 16S rDNA gene and thus could be more reliable in differentiating Serratia fonticola. A species-specific primer pair and probe were designed for quantitative real-time PCR detection of S. fonticola using gyrB as the target gene. Nine members of the Serratia family (representing nine Serratia species) were chosen to verify the specificity of the primers. Additionally, two species each of Salmonella and Klebsiella, and five other species belonging to five other genera of Enterobacteriaceae, were tested for primer cross-reaction. All the tested strains gave negative results. The limit of detection for S. fonticola using the gyrB gene was 100 copies per PCR reaction. This TaqMan PCR assay provided a specific, rapid, and sensitive method to detect S. fonticola based on its gyrB gene. |
| Databáze: | OpenAIRE |
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