Molecular detection of Pseudomonas stutzeri by duplex-PCR technique
Background and Objective: Duplex PCR is a widespread molecular biology technique that has the ability in specific and high sensitivity detection of microorganisms. This study was performed to evaluate the molecular identification of Pseudomonas stutzeri using duplex PCR. Methods: In this descriptive...
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| Format: | Article |
| Language: | fas |
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Golestan University of Medical Sciences
2019-01-01
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| Series: | مجله دانشگاه علوم پزشکی گرگان |
| Subjects: | |
| Online Access: | http://goums.ac.ir/journal/browse.php?a_code=A-10-1-1083&slc_lang=en&sid=1 |
| _version_ | 1818081951962103808 |
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| author | Roya Beytsayyah (Alavi Sharif) Fatemeh Haddadi Hossein Kamaladini Mirza Mohammad Reza Sharifmoghadam |
| author_facet | Roya Beytsayyah (Alavi Sharif) Fatemeh Haddadi Hossein Kamaladini Mirza Mohammad Reza Sharifmoghadam |
| author_sort | Roya Beytsayyah (Alavi Sharif) |
| collection | DOAJ |
| description | Background and Objective: Duplex PCR is a widespread molecular biology technique that has the ability in specific and high sensitivity detection of microorganisms. This study was performed to evaluate the molecular identification of Pseudomonas stutzeri using duplex PCR.
Methods: In this descriptive-laboratory study, Pseudomonas stutzeri ATCC 17588 bacteria was purchased from genetic resources center and after culturing the bacteria, DNA was extracted in the exponential growth phase using boiling method. Duplex PCR was carried out for specific identification of the bacteria subsequently. The primers were designed using catA and nirP gene sequences. Sensitivity and specificity of duplex PCR technique were investigated using 5 bacteria.
Results: The amplification of two bands of 512 bp and 249 bp for catA and nirP genes were observed, respectively. The specificity was 100% .The sensitivity of 0.048 ng/µL of genomic DNA was determined for catA and nirP genes, respectively.
Conclusion: Duplex-PCR molecular method with its sensitivity and proper feature and high potential for identification of Pseudomonas bacteria can be applied as a routine method in well-equipped laboratories by expert technician to identify suspicious cases. |
| first_indexed | 2024-12-10T19:14:22Z |
| format | Article |
| id | doaj.art-62a542b7e36b4729809fd69bcd16de69 |
| institution | Directory Open Access Journal |
| issn | 1562-4765 2008-4080 |
| language | fas |
| last_indexed | 2024-12-10T19:14:22Z |
| publishDate | 2019-01-01 |
| publisher | Golestan University of Medical Sciences |
| record_format | Article |
| series | مجله دانشگاه علوم پزشکی گرگان |
| spelling | doaj.art-62a542b7e36b4729809fd69bcd16de692022-12-22T01:36:38ZfasGolestan University of Medical Sciencesمجله دانشگاه علوم پزشکی گرگان1562-47652008-40802019-01-01204115122Molecular detection of Pseudomonas stutzeri by duplex-PCR techniqueRoya Beytsayyah (Alavi Sharif)0Fatemeh Haddadi1Hossein Kamaladini2Mirza Mohammad Reza Sharifmoghadam3 M.Sc in Genetics, Department of Biology, Faculty of Sciences, Zabol University, Zabol, Iran Assistant Professor, Department of Biology, Faculty of Sciences, Zabol University, Zabol, Iran Assistant Professor, Department of Biology, Faculty of Sciences, Zabol University, Zabol, Iran Assistant Professor, Department of Biology, Faculty of Sciences, Ferdowsi University, Mashhad, Iran Background and Objective: Duplex PCR is a widespread molecular biology technique that has the ability in specific and high sensitivity detection of microorganisms. This study was performed to evaluate the molecular identification of Pseudomonas stutzeri using duplex PCR. Methods: In this descriptive-laboratory study, Pseudomonas stutzeri ATCC 17588 bacteria was purchased from genetic resources center and after culturing the bacteria, DNA was extracted in the exponential growth phase using boiling method. Duplex PCR was carried out for specific identification of the bacteria subsequently. The primers were designed using catA and nirP gene sequences. Sensitivity and specificity of duplex PCR technique were investigated using 5 bacteria. Results: The amplification of two bands of 512 bp and 249 bp for catA and nirP genes were observed, respectively. The specificity was 100% .The sensitivity of 0.048 ng/µL of genomic DNA was determined for catA and nirP genes, respectively. Conclusion: Duplex-PCR molecular method with its sensitivity and proper feature and high potential for identification of Pseudomonas bacteria can be applied as a routine method in well-equipped laboratories by expert technician to identify suspicious cases.http://goums.ac.ir/journal/browse.php?a_code=A-10-1-1083&slc_lang=en&sid=1Pseudomonas stutzeriDuplex-PCRcatA genenirP gene |
| spellingShingle | Roya Beytsayyah (Alavi Sharif) Fatemeh Haddadi Hossein Kamaladini Mirza Mohammad Reza Sharifmoghadam Molecular detection of Pseudomonas stutzeri by duplex-PCR technique مجله دانشگاه علوم پزشکی گرگان Pseudomonas stutzeri Duplex-PCR catA gene nirP gene |
| title | Molecular detection of Pseudomonas stutzeri by duplex-PCR technique |
| title_full | Molecular detection of Pseudomonas stutzeri by duplex-PCR technique |
| title_fullStr | Molecular detection of Pseudomonas stutzeri by duplex-PCR technique |
| title_full_unstemmed | Molecular detection of Pseudomonas stutzeri by duplex-PCR technique |
| title_short | Molecular detection of Pseudomonas stutzeri by duplex-PCR technique |
| title_sort | molecular detection of pseudomonas stutzeri by duplex pcr technique |
| topic | Pseudomonas stutzeri Duplex-PCR catA gene nirP gene |
| url | http://goums.ac.ir/journal/browse.php?a_code=A-10-1-1083&slc_lang=en&sid=1 |
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