[Home ] [Archive]   [ فارسی ]  
:: Main :: About :: Current Issue :: Archive :: Search :: Submit :: Contact ::
Main Menu
Home::
Journal Information::
Indexing Sources::
Editorial Board::
Executive Members::
Articles Archive::
Instruction to Authors::
Peer-Review::
Contact Us::
Site Facilities::
::
Search in website

Advanced Search
Receive site information
Enter your Email in the following box to receive the site news and information.

Happy Persian New Year (Nowruz)


:: 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:   (6336 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]   (11864 Downloads)    
Type of Study: Original Articles | Subject: Microbiology
References
1. Dixit U, Shanker R. Detection of water-borne pathogens: culture plate to genomics. Indian J of Sci Technol. 2009 Nov; 2(11): 59-71.
2. Pindi PK, Srinath RR, Shanker AS. Novel approaches of genomic DNA isolation for identification of Cultivable Bacteria. Jundishapur J Microbiol. 2013 Dec; 6(10): e8339. doi: 10.5812/jjm.8339
3. Moolenaar RL, Crutcher JM, San Joaquin VH, Sewell LV, Hutwagner LC, Carson LA, et al. A prolonged outbreak of Pseudomonas aeruginosa in a neonatal intensive care unit: did staff fingernails play a role in disease transmission? Infect Control Hosp Epidemiol. 2000 Feb; 21(2): 80-5. doi: 10.1086/501739
4. Aidar M, Line SR. A simple and cost-effective protocol for DNA isolation from buccal epithelial cells. Braz Dent J. 2007; 18(2): 148-52.
5. Osmani Bojd M, Kamaladini H, Haddadi F, Vaseghi A. Thiolated AuNP probes and multiplex PCR for molecular detection of Staphylococcus epidermidis. Mol Cell Probes. 2017 Aug; 34: 30-36. doi: 10.1016/j.mcp.2017.04.006
6. Aydemir O, Aydemir Y, Ozdemir M. The role of multiplex PCR test in identification of bacterial pathogens in lower respiratory tract infections. Pak J Med Sci. 2014 Sep; 30(5): 1011-16. doi: 10.12669/pjms.305.5098
7. Iroha IR, Oji AE, Nwosu OK, Amadi ES. Antimicrobial Activity of Savlon®, Izal® and Z-Germicide® Against Clinical Isolates of Pseudomonas aeruginosa from Hospital Wards. European Journal of Dentistry and Medicine. 2011; 3: 32-35. doi: 10.3923/ejdm.2011.32.35
8. White DG, McDermott PF. Biocides, drug resistance and microbial evolution. Curr Opin Microbiol. 2001 Jun; 4(3): 313-37.
9. Brooks G, Carroll KC, Butel J, Morse S. Medical Microbiology (Jawetz, Melnick, & Adelberg's Medical Microbiology). 24th ed. New York: McGraw-Hill Medical. 2007; pp: 352-46.
10. Eriksen HM, Iversen BG, Aavitsland P. Prevalence of nosocomial infections in hospitals in Norway, 2002 and 2003. J Hosp Infect. 2005 May; 60(1): 40-5. doi: 10.1016/j.jhin.2004.09.038
11. Gilardi GL, Mankin HJ. Infection due to Pseudomonas stutzeri. NY State J Med. 1973 Dec; 73(23): 2789-91.
12. Priestley GS, Holland J, Marsh B, Wilson R. Pseudomonas stutzeri septicaemia in association with a bullous skin eruption. Anaesth Intensive Care. 1996 Dec; 24(6): 710-13.
13. Reisler RB, Blumberg H. Community-acquired Pseudomonas stutzeri vertebral osteomyelitis in a previously healthy patient: case report and review. Clin Infect Dis. 1999 Sep; 29(3): 667-69.
14. Rosenberg I, Leibovici L, Mor F, Block C, Wysenbeek AJ. Pseudomonas stutzeri causing late prosthetic valve endocarditis. J R Soc Med. 1987 Jul; 80(7): 457-59. doi: 10.1177/014107688708000719
15. Lebowitz D, Gürses-Ozden R, Rothman RF, Liebmann JM, Tello C, Ritch R. Late-onset bleb-related panophthalmitis with orbital abscess caused by Pseudomonas stutzeri. Arch Ophthalmol. 2001 Nov; 119(11): 1723-25.
16. Lalucat J, Bennasar A, Bosch R, García-Valdés E, Palleroni NJ. Biology of Pseudomonas stutzeri. Microbiol Mol Biol Rev. 2006 Jun; 70(2): 510-47. doi: 10.1128/MMBR.00047-05
17. Hernandez Duquino H, Rosenberg FA. Antibiotic-resistant Pseudomonas in bottled drinking water. Can J Microbiol. 1987 Apr; 33(4): 286-89.
18. Holmes B. Identification and distribution of Pseudomonas stutzeri in clinical material. J Appl Bacteriol. 1986 May; 60(5): 401-11.
19. Taneja N, Meharwal SK, Sharma SK, Sharma M. Significance and characterisation of pseudomonads from urinary tract specimens. J Commun Dis. 2004 Mar; 36(1): 27-34.
20. Kim MJ, Kim HY. Species identification of commercial jerky products in food and feed using direct pentaplex PCR assay. Food Control. 2017; 78: 1-6. http://dx.doi.org/10.1016/j.foodcont.2017.02.027
21. Shahbazi B, Narenji H. [Comparison of four methods of DNA extraction from gram-negative and gram-positive bacteria]. Zanko J Med Sci. 2014; 15 (45) :9-16. [Article in Persian]
22. Solà M, Riudavets J, Agustí N. Detection and identification of five common internal grain insect pests by multiplex PCR. Food Cntrol. 2018 Feb; 84: 246-54. https://doi.org/10.1016/j.foodcont.2017.08.002
23. Markoulatos P, Siafakas N, Moncany M. Multiplex polymerase chain reaction: a practical approach. J Clin Lab Anal. 2002; 16(1): 47-51.
24. Cladera AM, Bennasar A, Barceló M, Lalucat J, García-Valdés E. Comparative genetic diversity of Pseudomonas stutzeri genomovars, clonal structure, and phylogeny of the species. J Bacteriol. 2004 Aug; 186(16): 5239-48. doi: 10.1128/JB.186.16.5239-5248.2004
Send email to the article author


XML   Persian Abstract   Print


Download citation:
BibTeX | RIS | EndNote | Medlars | ProCite | Reference Manager | RefWorks
Send citation to:

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
URL: http://goums.ac.ir/journal/article-1-3543-en.html


Rights and permissions
Creative Commons License This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License.
Volume 20, Issue 4 (12-2018) Back to browse issues page
مجله دانشگاه علوم پزشکی گرگان Journal of Gorgan University of Medical Sciences
Persian site map - English site map - Created in 0.04 seconds with 38 queries by YEKTAWEB 4645