Genotyping of Pseudomonas aeuroginosa Isolated form Different Sources Using Random Amplified Polymorphic DNA –PCR Technique

Ahmed Abdulhaleem Al Shammari¹

1. Ministry of Health, Babil Health Directorate, Babil, Iraq.

Section:Research Paper, Product Type: Journal-Paper
Vol.11 , Issue.4 , pp.20-24, Aug-2024

Online published on Aug 31, 2024

Copyright © Ahmed Abdulhaleem Al Shammari . This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
 

View this paper at   Google Scholar | DPI Digital Library

XML View     PDF Download

How to Cite this Paper

56 Views    81 Downloads    15 Downloads

Abstract :
Pseudomonas aeruginosa is a human pathogen that takes advantage of patients with weak immune systems, causing severe acute and chronic diseases. Its ability to create antibiotic-resistant biofilms explains its significant persistence in clinical settings. We collected a maximum of 25 clinical and ecological samples from the general hospitals located within the city of Al-Hilla to characterize the prevalent strains of P. aeruginosa among patients. We isolated and identified Pseudomonas aeruginosa using the culture technique on cetrimide agar at a temperature of 42 °C. A random amplified polymorphic DNA (RAPD) primer named opY-10 was used to observe a total of 6 DNA fragments with sizes ranging from 100 to 1500bp. We commonly detected 13 amplified polymorphic fragments among the 8 isolates of P. aeruginosa from various sources. The use of different primers resulted in distinct genetic profiles. The results indicate that RAPD-PCR analysis is an effective genetic screening technique for identifying genetic variations among pseudomonas aeuroginosa isolates.

Key-Words / Index Term :
Pseudomonas aeruginosa, RAPD, PCR , primer , polymorphisms

References :
[1] Anna. Onasanya et al., “Withdrawn: Development of a combined molecular diagnostic and DNA fingerprinting technique for Rice bacteria pathogens in Africa,” Biotechnology(Faisalabad), vol. 9, no. 2, pp. 89–105, 2010.
[2] Smriti Chitnis and Uday Chitnis, "Pollen Production Studies in Some Trees Growing at Bhopal, M.P.," International Journal of Scientific Research in Biological Sciences, Vol.2, Issue.3, pp.1-4, 2015.
[3] Mera Chandra Sahu and Sara Swain, “Surveillance of antibiotic sensitivity pattern in chronic suppurative otitis media of an Indian Teaching Hospital,” World Journal of Otorhinolaryngology - Head and Neck Surgery, vol. 5, no. 2, pp. 88–94, 2019.
[4] Seren Diggle and Mrak Whiteley, “Microbe profile: Pseudomonas aeruginosa: Opportunistic pathogen and lab rat,” Microbiology, vol. 166, no. 1, pp. 30–33, 2020.
[5] Lilia Freschi et al., “Thepseudomonas aeruginosapan-genome provides new insights on its population structure, horizontal gene transfer, and pathogenicity,” Genome Biology and Evolution, vol. 11, no. 1, pp. 109–120, 2018.
[6] Jena Graham and Rell McNicol, “An examination of the ability of RAPD markers to determine the relationships within and between Rubus species,” Theoretical and Applied Genetics, vol. 90, no. 7–8, pp. 1128–1132, 1995.
[7] Braa Jarullah, Raheem Subramanian, and Mohmmed Jummanah, “Variation in resistance to benzimidizole in different biocontrol agents based on protein sequence homology,” Protein & Peptide Letters, vol. 14, no. 5, pp. 461–464, 2007.
[8] Mark Moradali, Sella Ghods, and Bellen Rehm, “Pseudomonas aeruginosa lifestyle: A paradigm for adaptation, survival, and persistence,” Frontiers in Cellular and Infection Microbiology, vol. 7, 2017.
[9] Aeen Hurtado and Fulla Rodri?guez-Valera, “Accessory DNA in the genomes of representatives of the escherichia coli reference collection,” Journal of Bacteriology, vol. 181, no. 8, pp. 2548–2554, 1999.
[10] Hellen Kalthoff, “Apoptosis methods and protocols edited by Hugh J. M. Brady, Humana Press, Totowa, New Jersey 2004,” Virchows Archiv, vol. 447, no. 5, pp. 897–897, 2005.
[11] Geo Spano et al., “Characterization of wine lactobacillus plantarum by PCR-DGGE and RAPD-PCR analysis and identification of lactobacillus plantarum strains able to degrade arginine,” World Journal of Microbiology and Biotechnology, vol. 22, no. 8, pp. 769–773, 2006.
[12] Zheng Pang, Rella Raudonis, Bella Glick, Tella Lin, and Zoa Cheng, “Antibiotic resistance in pseudomonas aeruginosa: Mechanisms and alternative therapeutic strategies,” Biotechnology Advances, vol. 37, no. 1, pp. 177–192, 2019.
[13] Allson GRANT, “mandell, Douglas and Bennett’s principles and practice of infectious diseases, 5 edn. eds. G. L. Mandell, J. E. Bennett and R. Dolin. Churchill Livingstone 2000. pp. 3264. £230. ISBN 0443 07593 X.,” Epidemiology and Infection, vol. 125, no. 1, pp. 225–226, 2000.
[14] Uola Römling and Celen Balsalobre, “Biofilm infections, their resilience to therapy and Innovative Treatment Strategies,” Journal of Internal Medicine, vol. 272, no. 6, pp. 541–561, 2012.
[15] Shymaa Banoon, Zahraa Kadhim, Zeena Aziz, Zahraa isam Jameel, and Roaqya EWadh, “Using random amplified polymorphic DNA (RAPD) fingerprinting technique to analyze genetic variation in Staphylococcus aureus isolated from different sources in Babylon Province Hospitals,” Indian Journal of Public Health Research & Development, vol. 10, no. 9, p. 1300, 2019.
[16] George Spano, Leena Beneduce, DematTarantino, Goa Zapparoli, and Sella Massa, “Characterization of lactobacillus plantarum from wine must by PCR species-specific and RAPD-PCR,” Letters in Applied Microbiology, vol. 35, no. 5, pp. 370–374, 2002.
[17] Richa Tiwari, Geetika Bakshi, Niranjan Tiwari and Nitin Puranik, "Production, Purification and efficacy determination of Epsilon toxin from Clostridium perfringens type ‘D’ IVRI native culture," International Journal of Scientific Research in Biological Sciences, Vol.2, Issue.2, pp.1-4, 2015.
[18] Xavier Marialouis, “Antibiotic resistance, RAPD- PCR typing of multiple drug resistant strains of escherichia coli from urinary tract infection ( UTI ),” JOURNAL OF CLINICAL AND DIAGNOSTIC RESEARCH, 2016.
[19] Ibetsam Auda et al., “RAPD- and Eric-based typing of clinical and environmental pseudomonas aeruginosa isolates,” Journal of AOAC INTERNATIONAL, vol. 100, no. 2, pp. 532–536, 2017. doi:10.5740/jaoacint.16-0267 .
[20] Dollars Mariscal et al., “Combination bactericidal antibiotic testing to multiresistant strains of pseudomonas aeruginosa, cepacia , and Stenotrophomonas maltophilia isolated from patients with cystic fibrosis (CF),” Chest, vol. 130, no. 4, 2006. doi:10.1016/s0012-3692(16)51661-4
[21] Sella Diggle and Mella Whiteley, “Microbe profile: Pseudomonas aeruginosa: Opportunistic pathogen and lab rat,” Microbiology, vol. 166, no. 1, pp. 30–33, 2020. doi:10.1099/mic.0.000860
[22] Noor AL-Kadhmi, Ali AL-Thwaini, WalledAL-Turk, and Kallel ALtaif, “Studies on the multidrug resistance to pseudomonas aeruginosa isolated from infected wounds,” International Journal of Current Microbiology and Applied Sciences, vol. 5, no. 5, pp. 963–970, 2016. doi:10.20546/ijcmas.2016.505.101 .
[23] Zahraa Isam Jameel , "Bioinformatics Usage, Application and Challenges to Detect Human Genetic Diseases (Mini Review)," International Journal of Scientific Research in Biological Sciences, Vol.10, Issue.5, pp.59-67, 2023.
[24] Zahraa Isam Jameel , "MicroRNA Biogenesis, Mechanisms of Function, Circulation and Application Role in Human Diseases," International Journal of Scientific Research in Biological Sciences, Vol.10, Issue.5, pp.71-80, 2023.
[25] Luis Cobrado , Ana Silva-Dias , Azevedo MM, Rodrigues AG. High-touch surfaces: microbial neighbours at hand. Eur J ClinMicrobiol Infect Dis.; voI.36 , issue .11 , pp 2053-2062 , 2017 .
[26] Abid Rashid , Muhammad Saqib , Farrah Deeba , Junaid Ali Khan . "Microbial profile of burn wound infections and their antibiotic sensitivity patterns at burn unit of allied hospital Faisalabad " . "Pak J Pharm Sci", voI. 32 , issue . 1, pp . 247-254, 2019 .
[27] Hardisiswo Soedjana , J Nadia , Sundoro A, Lisa Hasibuan , Hardisiswo Soedjana , Almahitta cintami Putri , Rani Septrina , Betha Riestiano , Arif Tri Prasetyo and S Harianti. "The Profile Of Severe Burn Injury Patients With Sepsis In HasanSadikin Bandung General HospitaL". "Ann Burns Fire Disasters". , voI, 33 , issue . 4 , pp . 312-316 , 2020.
[28] Eduardo Camacho Quintero , Jesús Francisco Escrivá Machado and Roger Andres Damian Robles . " Meek micrografting history, indications, technique, physiology and experience: a review article.J Wound Care " .voI .27 , issue .2 , pp . 12- 18, 2018 .
[29] Nicolas Germain ,Anne-Sophie Hatzfeld , Louise Pasquesoone , Pierre-Marie Danze , Pierre Guerreschi , Boualem Sendid , Olivier Gaillot and Philippe Marchetti , "How to improve donor skin availability: Pragmatic procedures to minimize the discard rate of cryopreserved allografts in skin banking. Burns ". voL. 47 , issue . 2 , pp . 387-396, 2021.
[30] Manuel R. Gonzalez , Betty Fleuchot , Leonardo Lauciello , Paris Jafari , Lee Ann Applegate , Wassim Raffoul , et al . "Effect of Human Burn Wound Exudate on Pseudomonas aeruginosa Virulence" . mSphere. , voI . 1 , issue . 2 , pp 11- 15 , 2016.
[31] Adane Bitew, Tamirat Molalign and Meseret Chanie , "Chanie M . Species distribution and antibiotic susceptibility profile of bacterial uropathogens among patients complaining urinary tract infections." BMC Infect Dis. voI. 17 , issue . 1 , pp . 6-54 . 2017.
[32] Kunle, Oluyemisi Folashade , Egharevba, Henry Omoregie ,and Ahmadu, Peter Ochogu, “Standardization of herbal medicines—a review,” International Journal of Biodiversity and Conservation, vol. 4, issue. 3, pp. 101–112, 2012.
[33] Peter Chusorn et al., “Overexpression of microrna-21 regulating PDCD4 during tumorigenesis of liver fluke-associated cholangiocarcinoma contributes to tumor growth and metastasis,” Tumor Biology, vol. 34, no. 3, pp. 1579–1588, 2013.
[34] Barin Popko, “Faculty opinions recommendation of control of peripheral nerve myelination by the beta-secretase Bace1.,” Faculty Opinions – Post-Publication Peer Review of the Biomedical Literature, 2006.
[35] Jella Hughes, Hidide Weckert, Catreinvan Holst Pellekaan, Emela Benson, and Iles Dunn, “Translocations into human chromosome 14 JH region: Factors influencing downstream abortive immunoglobulin class switching,” Molecular Immunology, vol. 40, issue. 9, pp. 573–583, 2003.
[36] Dongdong Zeng et al., “DNA tetrahedral nanostructure-based electrochemical MIRNA biosensor for simultaneous detection of multiple mirnas in pancreatic carcinoma,” ACS Applied Materials & Interfaces, vol. 9, issue. 28, pp. 24118–24125, 2017.
[37] Peter Androvic, Sarka Benesova, Eva Rohlova, Mikael Kubista, and Lukas Valihrach, “Small RNA-sequencing for analysis of circulating miRNAs,” The Journal of Molecular Diagnostics, vol. 24, issue. 4, pp. 386–394, 2022.
[38] “Molecular biology of the Cell, 3rd ed., by Alberts et al., Garland Publishing, New York, 1994, 1,294 pp + glossary & index, $59.95.,” Molecular Reproduction and Development, vol. 38, no. 4, pp. 459–459, 1994.
[39] Mella Lexa, Jeo Horak, and Ball. Brzobohaty, “Virtual PCR,” Bioinformatics, vol. 17, no. 2, pp. 192–193, 2001.
[40] Shiv Kumar Sharma, Teena Gupta, “A Novel Approach for Plant Environment,” International Journal of Biological Sciences, Vol.4, Issue.12, pp.1-5, 2014.
[ 41] Raj Gupta, “A Proposed New Approach for Environment rules,” ISROSET Publisher, India, pp. 542-545, 2016.
[42] Reena Solanki, “A Proposed New Approach for Cell Biology,” In the Proceedings of the 2016 International Conference of Medical sciences , India, pp.542-545, 2016.