The molecular detection of the causative agent of plague on the basis of the pla gene
Yersinia pestis, a gram-negative rod belonging to the Enterobacteriaceae family, is the causative agent of plague. Classical methods of detecting the organisms are time-consuming, expensive and dangerous. The aim of the study was to design a Real-time PCR assay on the basis of the pla gene of Yersin...
| Main Authors: | , |
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| Format: | Article |
| Language: | English |
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Kharazmi University
2020-01-01
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| Series: | یافتههای نوین در علوم زیستی |
| Subjects: | |
| Online Access: | http://nbr.khu.ac.ir/article-1-3084-en.html |
| _version_ | 1818325781553610752 |
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| author | Khatereh Kabiri Keivan Majidzadeh |
| author_facet | Khatereh Kabiri Keivan Majidzadeh |
| author_sort | Khatereh Kabiri |
| collection | DOAJ |
| description | Yersinia pestis, a gram-negative rod belonging to the Enterobacteriaceae family, is the causative agent of plague. Classical methods of detecting the organisms are time-consuming, expensive and dangerous. The aim of the study was to design a Real-time PCR assay on the basis of the pla gene of Yersinia pestis. In this research the Real- time PCR test was optimized by using special primers for targeting pla gene. After preparing 10-fold serial dilutions of the pla and their analysis by the assay, the last dilution showing a fluorescent signal was confirmed as the limit of detection (LOD). A standard curve based on the Ct values was depicted, so the assay was developed to quantify the target gene. The analytical specificity was determined by subjecting the genome of some control negative bacteria to the assay. In this experiment, negative control genomes did not show detectable signals in the assay. The last dilution of pla plasmid which showed a fluorescent signal was 4.5 fg. So, the lower detectable copy numbers of the gene in a 20 μl PCR reaction was calculated as 1×103. |
| first_indexed | 2024-12-13T11:49:56Z |
| format | Article |
| id | doaj.art-e669f4a255aa495988608c539134bcbf |
| institution | Directory Open Access Journal |
| issn | 2423-6330 2476-7115 |
| language | English |
| last_indexed | 2024-12-13T11:49:56Z |
| publishDate | 2020-01-01 |
| publisher | Kharazmi University |
| record_format | Article |
| series | یافتههای نوین در علوم زیستی |
| spelling | doaj.art-e669f4a255aa495988608c539134bcbf2022-12-21T23:47:24ZengKharazmi Universityیافتههای نوین در علوم زیستی2423-63302476-71152020-01-0164374381The molecular detection of the causative agent of plague on the basis of the pla geneKhatereh Kabiri0Keivan Majidzadeh1 Department of Microbiology, North Tehran Branch, Islamic Azad University, Tehran, Iran Tasnim Biotechnology Research Center (TBRC), AJA University of Medical Sciences, Tehran, Iran Yersinia pestis, a gram-negative rod belonging to the Enterobacteriaceae family, is the causative agent of plague. Classical methods of detecting the organisms are time-consuming, expensive and dangerous. The aim of the study was to design a Real-time PCR assay on the basis of the pla gene of Yersinia pestis. In this research the Real- time PCR test was optimized by using special primers for targeting pla gene. After preparing 10-fold serial dilutions of the pla and their analysis by the assay, the last dilution showing a fluorescent signal was confirmed as the limit of detection (LOD). A standard curve based on the Ct values was depicted, so the assay was developed to quantify the target gene. The analytical specificity was determined by subjecting the genome of some control negative bacteria to the assay. In this experiment, negative control genomes did not show detectable signals in the assay. The last dilution of pla plasmid which showed a fluorescent signal was 4.5 fg. So, the lower detectable copy numbers of the gene in a 20 μl PCR reaction was calculated as 1×103.http://nbr.khu.ac.ir/article-1-3084-en.htmlcontrol negativereal- time pcrspecificitystandard curveyersinia pestis |
| spellingShingle | Khatereh Kabiri Keivan Majidzadeh The molecular detection of the causative agent of plague on the basis of the pla gene یافتههای نوین در علوم زیستی control negative real- time pcr specificity standard curve yersinia pestis |
| title | The molecular detection of the causative agent of plague on the basis of the pla gene |
| title_full | The molecular detection of the causative agent of plague on the basis of the pla gene |
| title_fullStr | The molecular detection of the causative agent of plague on the basis of the pla gene |
| title_full_unstemmed | The molecular detection of the causative agent of plague on the basis of the pla gene |
| title_short | The molecular detection of the causative agent of plague on the basis of the pla gene |
| title_sort | molecular detection of the causative agent of plague on the basis of the pla gene |
| topic | control negative real- time pcr specificity standard curve yersinia pestis |
| url | http://nbr.khu.ac.ir/article-1-3084-en.html |
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