Phenotypic and Molecular Detection of Metallo-β –Lactamase Producing Pseudomonas aeruginosa Isolates From Different Clinical Infections in Erbil

Main Article Content

Bakhtyiar Sh. Azeez
Kamal I. Bakr

Keywords

P. aeruginosa, MBL, Antimicrobials, 16SrRNA, blaVIM, blaIMP

Abstract

Metallo-β-lactamase (MBL) producing Pseudomonas aeruginosa has been documented to be a critical nosocomial infection. Itwas continuous intrinsic and acquired resistance to a various group of antimicrobial agents and its resistance ability to develop multidrug resistance lead to a severe therapeutic problem. The study aimed to identify the molecular characterisation of clinical isolates of Metallo-β -lactamase P. aeruginosain Erbil hospitals. This study was carried out during the period from October 2017 to March 2018. A total of 300 clinical specimens were collected from patients (urine 124, wound 80, burns 40, bronchial wash 30, and sputum 26) aged 15-65 years attending Rizgary, West emergency, Erbil teaching hospitals. Out of 300 specimens, 50 isolates of P. aeruginosawere recovered and accounted for 16% of hospitalised infection isolates, the diagnosis of P. aeruginosaisolates was confirmed phenotypically and genotypically via the amplification of 16SrRNAgeneby using PCR technique. All isolates were tested toward the different class of antimicrobials by using agar diffusion method and VITEK 2 system. Levofloxacin, Norfloxacin, and Imipenem was the most effective antimicrobial, and most of the isolates were high resistance to (P, L,V, PI, R, CHL, E, B, A, N, TE, G, MEM, CEF, CTX, ATM).The lowest resistance was to IMP, LEV and NOR. Out of 50 of isolates,14 (28%) were found to produce MBL.16SrRNAwere used to confirm P. aeruginosa and blaVIM, blaIMP used to detect the MBL. All isolates were positive for 16SrRNA, while 12 (85%) and 8 (57%) were positive for blaVIM and blaIMP genes. In conclusion, the present study proved thatMetallo-β-lactamase is producing P. aeruginosaisolated had phenotypic characterisation which strongly correlated with according to genotypic characterisation. To our knowledge, this is the first attempt in Erbil city.

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References

Agnieszka E. Laudy, Patrycja RoÂg, Katarzyna Smolińska-KroÂ, MilenaCÂ miel, Alicja Søoczyńska, Jan Patzer, et al., (2017). Prevalence of ESBL-producing Pseudomonas aeruginosa isolates in Warsaw, Poland, detected by various phenotypic and genotypic methods. PLOS;1: 1-15.
Alla Salwa J. Al-Awadi, and Ahmed S. Mohammed (2014). Detection of Metallo-β-Lactamase Producing Pseudomonas aeruginosa Isolated from Public and Private Hospitals in Baghdad, Iraq. Acta Medica Iranica; 54(2): 107-113.
Ali Hadi Salih, (2016). Genetic and Phenotypic characterization of Pseudomonas aeruginosa Isolated from Inpatients in Baghdad hospitals. M.Sc. Thesis, College of Medicine, University of Al-Qadissiyah.
Al-Haidary CHH. (2010). Microbiological study of urinary tract infection, antibiotics susceptibility pattern and extended spectrum beta lactamase prevalence among children in Erbil city. M.Sc. Thesis, College of Medicine, Hawler Medical University.
Ami Varaiya, Nikhil Kulkarni, Manasi Kulkarni, Pallavi Bhalekar & Jyotsana Dogra. (2008). Extend-spectrum beta-lactamase-producing E. coli and Klebsiella in diabetic foot infections. J Med Res; 127: 398-402.
Amina N. AL-Thwani. (2013). Detection of β- lactam Genes in Pseudomonas aeruginosa Isolates in Some Hospitals in Bagdad Governorate by using Multiplex PCR. Journal of Babylon University; special Issue: 1-10.
Arciola CR, Baldassarri L, Montanaro L. (2001). Presence of icaA and icaD genes and slime production in a collection of staphylococcal strains from catheter associated infections. Journal of clinical microbiology; 29(6): 2151-2156.
Asmaa A. AL-Kaisse. Amina N. AL-Thwani. Rabab Q. AL-Segar. (2015). PCR Detection of Some ESBLs (bla) Genes in Pseudomonas aeruginosa Isolated from Burn’s Units in Bagdad Hospitals. Journal of Biotechnology Research Center; 9(2): 74-80.

Azhar A.Neamah. (2017). Molecular Detection of virulence factor genes in pseudomonas aeruginosa isolated from human and animals in Diwaniya province. Kufa Journal for Veterinary Medical Sciences; 8(1): 218-229.
Clare F, Lisa L, and Anton Y P. (2006). Phenotypic Detection of carbapenem-Susceptible Metallo β-lactamase Producing Gram-negative Bacilli in the Clinical Laboratory. Journal of Clinical Microbiology; 44(9): 3139-3144.
Clinical and Laboratory Standards Institute. (2017). Performance standards for antimicrobial susceptibility testing. 27th ed. Informational Supplement. CLSI Document. Wayne P A. USA. Clinical and Laboratory Standards Institute.
David P. Speert, Susan W. Farmer, Maureen E. Campbell, James M. Musser, Robert K. Selander, and Susan Kuo. (1990). Conversion of Pseudomonas aeruginosa to the Phenotype Characteristic of Strains from Patients with Cystic Fibrosis. Journal of Clinical Microbiology; 28(2):188-194.
Delissable F, Aibile Ceuvas CF. (2004). Comparison of antibiotic susceptibility and plasmid content, between biofilm producing and non-producing clinical isolates of Pseudomonas aeruginosa. Journal Antimicrobial agents;24(4): 405-408.
Drieux L, F. Brossier, W. Sougakoff and V. Jarlier. (2008). Phenotypic detection of extended-spectrum b-lactamase production in Enterobacteriaceae: review and bench guide. Clin Microbiol Infect; 14(1): 90-103.
Eigner U, Schmid A, Wild U, Bertsch D and Fahr A M. (2005). Analysis of the comparative workflow and performance characteristics of the VITEK2 and Phoenix systems. Journal of Clinical Microbiology; 43: 3829-3834.
Ekrem K, Rokan DK (2014). Antibiotic susceptibility patterns of Pseudomonas aeruginosa strains isolated from various clinical specimens. Sky J. Microbiology. Res. 2(2):13-17.
Firdous R, Ahmed S, Chaudhary S A, Ahmad A and Akhtar N. (2011). Evaluation of Resistance in Clinical Isolates of E. coli, S, aureus, and Pseudomonas aeruginosa against β-lactam Antibiotics and Gentamycin. Journal of Rawalpindi Medical College (JRMC); 15 (1) 6-9.
Hallem H, Tarrad JK, Banyan IA. (2011). Isolation of Pseudomonas aeruginosa from clinical cases and environmental samples, and analysis of its antibiotic resistant spectrum at Hilla teaching hospital. Medical Journal of Babylon; 8(4): 45-52.
Hussam al-Jaafari. (2016). Antibiotic susceptibility of P. aeruginosa isolated from burns and wounds of patients, Iraq. Journal of Environmental Microbiology. 4 (1) 170-175.
Hussen S Q. (2010). Microbiological study of burn wound infection, Antibiotics susceptibility pattern and Beta lactamase prevalence in Erbil city. M.Sc. thesis. Erbil: Hawler Medical University. College of Medicine.
Johann D. D. Pitout, Daniel B. Gregson, Laurent Poirel, Jo-Ann McClure, Phillip Le, and Deirdre L. Church. (2005). Detection of Pseudomonas aeruginosa Producing Metallo-β-Lactamases in a Large Centralized Laboratory. Journal of Clinical Microbiology; 43(7): 3129-3135.

Mai M. Zafer, Mai M. Zafer, Hadir A. El-Mahallawy, Magdy A. Amin, and Mohammed Seif El-Din Ashour. (2014) Antimicrobial Resistance Pattern and Their Beta-Lactamase Encoding Genes among Pseudomonas aeruginosa Strains Isolated from Cancer Patients. BioMed Research International; 14(8): 1-8.
Mariappan Shanthi, Uma Sekar, Arunagiri Kamalanathan, Balarman Seker. (2014). Detection of New Delhi metallo beta lactamase-1 (NDM-1) carbapenemase in Pseudomonas aeruginosa in a single centre in southern India. IJMR; 140(40): 546-550.
Mohammed A. Hamod. (2015). Rapid High Specific Method for the Detection of Pseudomonas fluorescens. American Journal of Microbiological Research; 3(5): 160-164.
Produção de metalo-b-lactase de linhagens de Pseudomonas aeruginosa isoladas em hospitais do Recife. (2005). Metallo-b-lactamase producing Pseudomonas aeruginosa strains isolated in hospitals in Recife, PE, Brazil. Brazilian Journal Of Microbiology; 36(2): 111-118.
Robert E. W. Hancock. (1998). Resistance Mechanisms in Pseudomonas aeruginosa and Other Nonfermentative Gram-Negative Bacteria. Clinical Infectious Diseases; 27(10): 93-98.
Ryan, G.; Singh, M.; Dwan, K. (2011). Inhaled antibiotics for long-term therapy in cystic fibrosis. Cochrane Database Syst. Rev;16, CD00102.
Shukriyah S. S. (2013). Antibiotic resistance studies and curing analysis by ascorbic acid in Pseudomonas aeruginosa. Ph.D. Thesis, College of Medicine, Hawler Medical University.
Smet A., Martel A., Persoons D, Dewulf J, Heyndrickx M, Herman L, Haesebrouck F. & Butaye P. (2009). Broad-spectrum β-lactamases among Enterobacteriaceae of animal origin: molecular aspects, mobility and impact on public health. FEMS Microbiology Reviews; 34: 295-316.
Sunite A. Ganju, Ramesh Chand Guleria, Suruchi Bhagra, Anil K. Kanga. (2016). Screening for metallo-β-lactamase producing Pseudomonas aeruginosa in clinical isolates in a tertiary care hospital in North India. Medical Journal of Dr. D.Y. Patil University; 8(3): 334-336.
Tanzinah Nasrin, Md. Shariful Alam Jilani, Lovely Barai, J. Ashraful Haq. (2010). Metallo-ß-Lactamase Producing Pseudomonas species in a Tertiary Care Hospital of Dhaka City. Bangladesh J Med Microbiol; 4(1): 43-45.
Yoshichika Arakawa, Naohiro Shibata, Keigo Shibayama, Hiroshi Kurokawa, Tetsuya Yagi, Hiroshi Fujiwara, and Masafumi Goto. (2000). Convenient Test for Screening Metallo-β-Lactamase-Producing Gram-Negative Bacteria by Using Thiol Compounds. Journal of Clinical Microbiology; 38(1): 40-43.