Molecular analysis of clinical isolated Pseudomonas aeruginosa by ERIC PCR

Sura Abdul-Aziz Najem; Ebtehal E. Shubbar

doi:10.53730/ijhs.v6nS1.7203

Authors

Sura Abdul-Aziz Najem
suraa.albohommedy@student.uokufa.edu.iq
Laboratory Investigations Department, Faculty of science, University of Kufa, Iraq
Ebtehal E. Shubbar Laboratory Investigations Department, Faculty of science, University of Kufa, Iraq

Keywords:

ERIC-PCR, Pseudomonas aeruginosa, Molecular analysis

Abstract

Pseudomonas aeruginosa is a significant mortality factor due to nosocomial infections in humans. P. aeruginosa has been known with severe infections, high incidence, and multiple drug resistance The present study aims to rapidly diagnose and biotype the isolates of P. aeruginosa isolated from human infections in intensive care units, Al-Sadder Medical City, Al-Hakeem General Hospital, and the burn center in Najaf city by ERIC-PCR. From 156 clinical sample 23 P. aeruginosa were isolated including burn 17/77 (22%), 2/21wound (9.5%), 3/43 urine (6.9%), 1/15 throat swab(6.6) between August 2021 to February 2022. After phenotypic confirmation, isolates were examined by PCR for molecular confirmation Enterobacterial Repetitive Intergenic Consensus (ERIC) PCR were done on 24 isolates. In ERIC-PCR Genetic analysis of P. aeruginosa isolates depending on ≥ 75% similarity revealed the presence of 8 different cluster as figure 2, four of them with single isolate other two clusters with two similar isolates while the two other clusters contained seven identical isolates profile each.

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References

Abdelrahman, D.N., Taha, A.A., Dafaallah, M.M., Mohammed, A.A., El Hussein, A.R.M., Hashim, A.I., Hamedelnil, Y.F. and Altayb, H.N., 2020. β-lactamases (bla TEM, bla SHV, bla CTXM-1, bla VEB, bla OXA-1) and class C β-lactamases gene frequency in Pseudomonas aeruginosa isolated from various clinical specimens in Khartoum State, Sudan: a cross sectional study. F1000Research, 9.

Asgarani, E., Ghashghaei, T., Soudi, M.R. and Alimadadi, N., 2015. Enterobacterial repetitive intergenic consensus (ERIC) PCR based genetic diversity of Xanthomonas spp. and its relation to xanthan production. Iranian journal of microbiology, 7(1), p.38.

Auda, I.G., Al-Kadmy, I.M., Kareem, S.M., Lafta, A.K., A'Affus, M.H.O., Khit, I.A.A., Al Kheraif, A.A., Divakar, D.D. and Ramakrishnaiah, R., 2017. RAPD-and ERIC-based typing of clinical and environmental Pseudomonas aeruginosa isolates. Journal of AOAC International, 100(2), pp.532-536.

Badamchi, A., Masoumi, H., Javadinia, S., Asgarian, R. and Tabatabaee, A., 2017. Molecular detection of six virulence genes in Pseudomonas aeruginosa isolates detected in children with urinary tract infection. Microbial pathogenesis, 107, pp.44-47.

Bel Hadj Ahmed, A., Salah Abbassi, M., Rojo-Bezares, B., Ruiz-Roldán, L., Dhahri, R., Mehri, I., Sáenz, Y. and Hassen, A., 2020. Characterization of Pseudomonas aeruginosa isolated from various environmental niches: New STs and occurrence of antibiotic susceptible “high-risk clones”. International Journal of Environmental Health Research, 30(6), pp.643- 652..

Dawod, R.E., Studies on Pseudomonas aeruginosa infection in hatcheries and chicken Rehab E. Dawod, Wafaa A. Abd El-Ghany** and Khadra M. Soliman.

El-Badawy, M.F., Alrobaian, M.M., Shohayeb, M.M. and Abdelwahab, S.F., 2019. Investigation of six plasmid-mediated quinolone resistance genes among clinical isolates of pseudomonas: a genotypic study in Saudi Arabia. Infection and Drug Resistance, 12, p.915..

Harper, D.R., Parracho, H.M., Walker, J., Sharp, R., Hughes, G., Werthén, M., Lehman, S. and Morales, S., 2014. Bacteriophages and biofilms. Antibiotics, 3(3), pp.270-284.

Hematzadeh, A. and Haghkhah, M., 2021. Biotyping of isolates of Pseudomonas aeruginosa isolated from human infections by RAPD and ERIC-PCR. Heliyon, 7(9), p.e07967.

Hornef, M.W., Roggenkamp, A., Geiger, A.M., Hogardt, M., Jacobi, C.A. and Heesemann, J., 2000. Triggering the ExoS regulon of Pseudomonas aeruginosa: a GFP-reporter analysis of exoenzyme (Exo) S, ExoT and ExoU synthesis. Microbial pathogenesis, 29(6), pp.329-343.

Li, W., Raoult, D. and Fournier, P.E., 2009. Bacterial strain typing in the genomic era. FEMS microbiology reviews, 33(5), pp.892-916.

Mikkelsen, H., McMullan, R. and Filloux, A., 2011. The Pseudomonas aeruginosa reference strain PA14 displays increased virulence due to a mutation in ladS. PloS one, 6(12), p.e29113.

Ojeniyi, B. and Høiby, N., 1991. Comparison of different typing methods of Pseudomonas aeruginosa. In Pseudomonas aeruginosa in Human Diseases (Vol. 44, pp. 13-22). Karger Publishers.

PangZ, R. and GLICK, B., 2019. Antibioticresistancein Pseudomonasaeruginosa: mechanismsand alternativetheraG peuticstrategies. BiotechnolAdv, 37(1), p.177G192.

Preethirani, P.L., Isloor, S., Sundareshan, S., Nuthanalakshmi, V., Deepthikiran, K., Sinha, A.Y., Rathnamma, D., Nithin Prabhu, K., Sharada, R., Mukkur, T.K. and Hegde, N.R., 2015. Isolation, biochemical and molecular identification, and in-vitro antimicrobial resistance patterns of bacteria isolated from bubaline subclinical mastitis in South India. PLoS One, 10(11), p.e0142717.

Ranjbar, R., Tabatabaee, A., Behzadi, P. and Kheiri, R., 2017. Enterobacterial repetitive intergenic consensus polymerase chain reaction (ERIC-PCR) genotyping of Escherichia coli strains isolated from different animal stool specimens. Iranian journal of pathology, 12(1), p.25.

Salas-Pacheco, J.M., Setlow, B., Setlow, P. and Pedraza-Reyes, M., 2005. Role of the Nfo (YqfS) and ExoA apurinic/apyrimidinic endonucleases in protecting Bacillus subtilis spores from DNA damage. Journal of bacteriology, 187(21), pp.7374-7381.

Sallman, R.S., Hussein, S.S. and Ali, M.R., 2018. ERIC-PCR Typing, RAPD-PCR Fingerprinting and Quorum Sensing Gene Analysis of Pseudomonas aeruginosa Isolated from Different Clinical Sources. Al-Mustansiriyah Journal of Science, 29(2), pp.50-62.

Sorkh, M. A. G., Shokoohizadeh, L., Rashidi, N., & Tajbakhsh, E. (2017). Molecular analysis of Pseudomonas aeruginosa strains isolated from burn patients by repetitive extragenic palindromic-PCR (rep-PCR). Iranian Red Crescent Medical Journal, 19(4), e43508

Stehling, E.G., Leite, D.S. and Silveira, W.D., 2010. Molecular typing and biological characteristics of Pseudomonas aeruginosa isolated from cystic fibrosis patients in Brazil. The Brazilian Journal of Infectious Diseases, 14(5), pp.462-467.

Syrmis, M.W., O'Carroll, M.R., Sloots, T.P., Coulter, C., Wainwright, C.E., Bell, S.C. and Nissen, M.D., 2004. Rapid genotyping of Pseudomonas aeruginosa isolates harboured by adult and paediatric patients with cystic fibrosis using repetitive-element-based PCR assays. Journal of Medical Microbiology, 53(11), pp.1089-1096.

Todar, K., 2008. Todar’s online textbook of microbiology. University of Wisconsin-Madison Department of Bacteriology.

Uchiyama, J., Suzuki, M., Nishifuji, K., Kato, S.I., Miyata, R., Nasukawa, T., Yamaguchi, K., Takemura-Uchiyama, I., Ujihara, T., Shimakura, H. and Murakami, H., 2016. Analyses of short-term antagonistic evolution of pseudomonas aeruginosa strain pao1 and phage kpp22 (myoviridae family, pb1-like virus genus). Applied and environmental microbiology, 82(15), pp.4482-4491.

Van Delden, C. and Iglewski, B.H., 1998. Cell-to-cell signaling and Pseudomonas aeruginosa infections. Emerging infectious diseases, 4(4), p.551..

Versalovic, J., Koeuth, Y. and Lupski, J.R., 1991. Polymerase chain reaction with consensus sequences primers is a powerful method for typing bacteria. Nucl. Acids Res, 19, pp.6823- 6831.

Wassenaar, T.M. and Newell, D.G., 2000. Genotyping of Campylobacter spp. Applied and Environmental Microbiology, 66(1), pp.1-9.

Wilson, L.A. and Sharp, P.M., 2006. Enterobacterial repetitive intergenic consensus (ERIC) sequences in Escherichia coli: Evolution and implications for ERIC-PCR. Molecular biology and evolution, 23(6), pp.1156-1168.