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:: Volume 20, Issue 4 (12-2018) ::
J Gorgan Univ Med Sci 2018, 20(4): 115-122 Back to browse issues page
Molecular detection of Pseudomonas stutzeri by duplex-PCR technique
Roya Beytsayyah (Alavi Sharif)1 , Fatemeh Haddadi * 2, Hossein Kamaladini3 , Mirza Mohammad Reza Sharifmoghadam4
1- M.Sc in Genetics, Department of Biology, Faculty of Sciences, Zabol University, Zabol, Iran
2- Assistant Professor, Department of Biology, Faculty of Sciences, Zabol University, Zabol, Iran , fatemeh.haddadi@uoz.ac.ir
3- Assistant Professor, Department of Biology, Faculty of Sciences, Zabol University, Zabol, Iran
4- Assistant Professor, Department of Biology, Faculty of Sciences, Ferdowsi University, Mashhad, Iran
Abstract:   (8010 Views)
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.
Keywords: Pseudomonas stutzeri, Duplex-PCR, catA gene, nirP gene
Full-Text [PDF 287 kb]   (15903 Downloads)    
Type of Study: Original Articles | Subject: Microbiology
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Beytsayyah (Alavi Sharif) R, Haddadi F, Kamaladini H, Sharifmoghadam M M R. Molecular detection of Pseudomonas stutzeri by duplex-PCR technique. J Gorgan Univ Med Sci 2018; 20 (4) :115-122
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Volume 20, Issue 4 (12-2018) Back to browse issues page
مجله دانشگاه علوم پزشکی گرگان Journal of Gorgan University of Medical Sciences
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