Published Journal Articles
Prevalence of Metallo-β-Lactamase Producing Pseudomonas aeruginosa in wound infections in Duhok city, Iraq.
International Journal of Research in Medical Sciences (IJRMS) (Issue : 4) (Volume : 4)
Background: Pseudomonas aeruginosa is common pathogen causing nosocomial infection. Acquired drug resistance and Metallo-β-lactamases (MBL) production have recently emerged as one of the most worrisome resistance mechanism that hydrolyze all beta-lactam antibiotics including penicillins, cephalosporins and carbapenems, with the exception of aztreonam. The aim was to find out the prevalence of multi drug resistant (MDR) and Metallo-β-lactamase (MBL) positive isolates of P. aeruginosa in wounds samples which are a serious concern. Methods: Pseudomonas aeruginosa strains were obtained by standard isolation and identification techniques from 307 wound samples of hospital. Strains were then subjected to susceptibility testing for anti-pseudomonas drugs as per Clinical and Laboratory Standards Institute (CLSI) guidelines. Carbapenems resistant strains were selected for the detection of MBL enzyme production by disc potentiation test. Production of MBL was confirmed by enhancement of inhibition zone around imipenem and meropenem discs impregnated with EDTA, as compared to discs without EDTA. Results: Amongst the 71 isolates of P. aeruginosa, 62(87.3%) isolate were imipenem-sensitive, while 9(12.7%) isolates were found to be imipenem resistant and MBL producers. Very high resistance to antibiotics was recorded amongst MBL producers’ P. aeruginosa compared with non-MBL imipenem-sensitive strains. Conclusion: Study indicates that, surveillance for the detection of MBL is necessary. The rapid dissemination of MBL producers is worrisome and necessitates the implementation of proper and judicious selection of antibiotics especially carbapenem.
Molecular Characterization of Some Virulence Genes and Antibiotics Susceptibility Pattern among Uropathogenic Escherichia coli Isolated From Patient in Zakho City/Iraq
zanco journal of pure and applied sciences (Issue : 32) (Volume : 2)
Uropathogenic Escherichia coli (UPEC) is one of the most causative agents which causing urinary tract infections (UTIs) in humans. This study involved the prevalence of the virulence genes among UPEC isolated from patients in various hospitals in Zakho city from July 2018 until January 2019 and their susceptibility to different commonly used antimicrobial agent against UPEC. The different culture media were used for the identification of Escherichia coli (E. coli). Out of 400 samples, 141 (35.25%) strains of UPEC were isolated from enrolled patients. The antibiotic susceptibility toward different antibiotics was varied among the isolates. Imipenem was the most potent antibiotic with a resistant rate of only 2.84%. While the isolates were resistant to most screened antibiotics, with the highest rate 96.45% to Amoxicillin/ clavulanic. The resistant rates decreased toward other antibiotics at rates varied from 93.62% for Amoxicillin to 43.97% for Norfloxacin. Fifty-seven isolates were selected for PCR analysis, according to the resistance of E.coli to various antibiotics. The selected samples were successfully amplified for E. coli identification by producing a single band of a target uidA gene. In this study, the virulence related genes were detected in only 35 (61.40%) isolates out of 57 isolates. The distribution of the virulence related genes that included; afa, sfa, hly, cnf and pai were 28.07%, 17.54%, 26.32%, 22.81% and 22.81%, respectively. The study highlight that multidrug resistance UPEC harbors multiple virulence genes circulating in this settin
Molecular differentiation and determination of multi-drug resistant isolates of Pseudomonas species collected from burn patients in Kurdistan Region, Iraq
Zanco Journal of Medical Sciences (Issue : 22) (Volume : 2)
Background and objective: Pseudomonas aeruginosa is very a well-documented nosocomial and opportunistic microorganism, a little challenge is being present with the identiﬁcation of such pathogen. This study aimed to identify Pseudomonas on genus and species levels by conventional PCR and determine multi-drug resistant isolates.
Distribution of extended spectrum β-lactamase genes among Proteus mirabilis isolated from clinical specimens in Duhok city, Kurdistan region, Iraq
Science Journal of University of Zakho (Issue : 5) (Volume : 1)
Extended Spectrum Beta Lactamase-among Proteus mirabilis strains recorded high incidence leaving few therapeutic options of potential infections. The purpose of current study was to assess the prevalence of antibiotic resistance among Extended Spectrum Beta-Lactamases (ESBL) producing P. mirabilis, in addition to molecular characterization of the ESBL gene-types using PCR. All isolates were fully identified, checked for antibiotic susceptibility and ESBL production using double disk synergy phenotypic method. Positive ESBL-producing isolates were subjected to PCR assay using specific primers for detection of CTX-M, TEM, and SHV genes. The majority of the isolates exhibited absolute susceptibility (100%) to both meropenem and ertapenem and high susceptibility (95%) to imipenem, while co-resistance were expressed toward cefotaxime, ceftazidime, ceftriaxone and other non-lactam antibiotics. Out of 37 isolates, 21 (57%) were ESBL-producers and using a double-disc synergy test (DDST). Using molecular-based PCR, CTX-M (81%), TEM (57%) and SHV (24%) were determined among ESBL-positive. CTX-M was predominant and circulating among phenotypic multiple resistant strains. Moreover, the coexistence of CTX-M and TEM gene was a more frequent combination. The study highlighted the increasing levels of low antibiotic susceptibility among P. mirabilis harbored ESBL genes at Duhok city and also confirms that a high level of blaCTX-M-positive ESBL isolates is circulating in this area.
Molecular Detection of Virulence Factors of Enterococcus Faecalis Isolated From Urine Samples in Duhok City, Kurdistan Region/Iraq.
Science Journal of University of Zakho (Issue : 4) (Volume : 1)
Enterococcus faecalis is one of the leading causes of many infections and mainly urinary tract infections. This pathogen developed high resistance to multiple antibiotics and it harbor many virulence factors genes. This study aimed to determine the antibiotic resistance patterns and screening for some virulence factor genes of E. faecalis isolated from urinary tract infection. Urine samples were collected from 788 outpatient’s clinic having clinical signs of UTI that visited Azadi Teaching Hospital in Duhok city. Urine samples were cultured on bacteriological media and isolated colonies identified using standard bacteriological methods. Antibiotic susceptibility was performed by Kirby Bauer test. All isolates were subjected to species-specific PCR assay for confirmatory identification followed by targeting virulence genes. Twenty five isolates of E. faecalis were detected and confirmed by species-specific PCR assay that expressed high antibiotic-resistance to many selected drugs except norfloxacin, penicillin and ampicillin. The most prevalent genes among all isolates were cpd genes followed by asa1, ace, esp, and gelE. Bearing of virulence genes combination were more frequent among multiple-antibiotic resistant strains. This study highlighted on E. faecalis as causes of UTI in Duhok city that showed multiple resistances to common antibiotics and harboring more than one virulence gene.
STUDY GENETIC RELATIONSHIP AMONG ENTEROCOCCUS FAECALIS STRAINS COLLECTED FROM URINE HARBORED DIFFERENT VIRULENCE PROFILES USING ERIC-PCR ASSAY
International Journal of Chemical and Biomolecular Science (Issue : 2) (Volume : 2)
Twenty five isolates of Enterococcus faecalis have been previously identified and characterized were subjected to ERIC-PCR analysis in order to study the genetic relationship with regarding to their virulence profile. Nine virulence profiles were identified according to the presence/absence of five virulence factors; asa1, gelE, esp, cpd and ace. Most isolates (28%) were belonged to Virprofile1 asa1+, gelE+, esp+, cpd+, ace+ followed by Virprofile3 asa1+, gelE-, esp+, cpd+, ace+(20%), Virprofile4 asa1+, gelE+, esp-, cpd+, ace-accounting 16%, while virprofiles5 asa1+, gelE-, esp-, cpd+, ace+, virprofile6 asa1-, gelE+, esp+, cpd+, ace+ and virprofile8 asa1+, gelE-, esp+, cpd+, ace-representing 8%, virprofile2 asa1-, gelE-, esp-, cpd-, ace+, virprofile7 asa1+, gelE+, esp+, cpd+, ace-and virprofile9 asa1+, gelE+, esp-, cpd+, ace+ were 4%. ERIC-PCR analysis divided isolates into two main clusters named; cluster A accounting 28% which further classified into groups; 8% isolates were belonged to A1 and 20% were belonged to A2. Most isolates were belonged to cluster B accounting 72%. This cluster was involved two groups; six isolates (33.3%) were belonged to B1 while 66.6% of isolates were assigned as B2. However, no relationship was found between virulence profiles with phylogentic groups of the isolates.
Molecular Characterization of Extended Spectrum B-lactamase Producing Escherichia coli Isolated from Urine in Kurdistan Region-Iraq
International Journal of Chemical and Biomolecular Science (Issue : 2) (Volume : 2)
Molecular characterization of ESBL-related bla genes including blaTEM, blaSHV, and blaCTX-M has been performed for Escherichia coli isolated from urine and collected from three cities in Kurdistan/region/Iraq (Erbil, Sulymani and Duhok). One hundred sixty nine isolates of E. coli have been identified and their production of ESBLs enzymes have been determined using phenotypic methods. All these isolates were successfully amplified producing a single band of the uidA locus in all strains with a molecular weight of about 670bp in order to confirm at molecular level that all these isolates were E. coli. One hundred sixty ESBL E. coli isolates out of 169 appeared to have one or more ESBLs genes accounting for 94.7%. CTX-M constituted the high prevalent type of ESBLs genes compared to the others represented by 94.1% of all isolates in all the three cities of Kurdistan region followed by TEM and SHV in a percentages of 43.8% and 2.5%, respectively. In Duhok, TEM showed the higher prevalence (60.8%) in comparison to the other two cities in percentages of 36.2% for Sulaimania while Erbil represented by 25%. Furthermore, it was clear that SHV type of ESBLs had the lower prevalence of all types and there were only four isolates out of 160 appeared to carry this type of gene representing 2.5%. The presence and/or absence of the three genes in all isolates were also investigated and it was shown that 86/160 isolates (53.75%) had the CTX-M gene only while the rest of genes were lacking. Moreover, 69/160 isolates had both CTXM and TEM. Interestingly, 3/160 harbored all three involved genes. The isolates characterized by the presence only TEM gene and those that had both CTX-M and SHV, shared the same percentage (0.6%). after taking sequencing of the PCR product of studied genes for 12 E coli isolates into consideration, it was obvious that all the PCR products of CTX-M were belonged to type CTXM-15; while TEM-1 type appeared predominant among all sequences PCR product for TEM gene.. Finally, from the three isolates which revealed positive PCR amplification for the SHV gene, two isolates showed 100% similarity to the SHV-12 genome type while the rest single isolate was similar (99%) to SHV11.
Multilocus Sequence Typing of Klebsiella Pneumoniae Producing Extended Spectrum Β-Lactamases Isolated From Kurdistan Region, Iraq.
Science Journal of University of Zakho (Issue : 2) (Volume : 2)
This study was purposed to sequence analysis of ESBLs genotype of K pneumoniae using partial sequence and Multilocus sequence typing (MLST). A total of 275 K. pneumoniae isolatesinvolved three general hospitals in Duhok, Erbil, and Sulymania, from September 2010 to June 2011. The Minimum Inhibitory Concentration (MIC) was measured by Phoenix system that confirmed 187 ESBL producing isolates followed by the Double Disk Synergy Test (DDST). Then, 12 isolates were selectedaccording to sample diversity, high resistancy to β-lactam and cephalosporins and harboring combination of three genes (TEM, SHV, and CTX-M). Partial sequence analysis of TEM; showed two different genotypes regarding blaTEM as 9 isolates (75%) from different samples (wound, sputum and blood) from three provinces harbor TEM-1 gene and 3 isolates (25%) only from urine in three provinces harbor TEM-198 gene. SHV analysis revealed characterization of selected isolates into six different genotypes. The common genotype was blaSHV-11 involved five isolates from sputum and blood in Erbil and Sulymania provinces, and wound in Duhok province. Only one genotype as all 12 isolates (100%) from different samples and different provinces was found harbored CTX-M-15 gene. Multilocus sequence typing (MLST) study performed using seven housekeeping genes (gapA, infB, mdh, pgi, phoE, rpoB and tonB). A total of 8 different sequence types (STs) were identified; ST11 was dominant sequence type, accounting 41.6%(5 isolates) and was harboring combination of TEM-1, SHV-11 and CTX-15 genes.
Bacteriological and Molecular Characterization of Extended Spectrum Β-Lactamases in Clinical Isolates of Klebsiella Pneumoniae Isolated From Kurdistan Region, Iraq.
Science Journal of University of Zakho (Issue : 1) (Volume : 1)
A total of 275 clinical isolates of Klebsiella pneumoniae were collected from three general hospitals in Duhok, Erbil, and Sulymania, during the period September 2010 to June 2011. The Minimum Inhibitory Concentration (MIC) of these isolates was measured using the Gram-negative susceptibility card (GNC) of Phoenix system. Only 187 ESBL producing K. pneumoniae isolates were detected by this system. These isolates were confirmed as 100% ESBLs producers by the Double Disk Synergy Test (DDST). All 187 K. pneumoniae isolates were 100% resistant to ampicillin, cefazolin, cefepime, ceftriaxone, cefotaxime, cefuroxime, ceftazidime, and aztreonam. These isolates showed different percentages of resistance 81.8%, 68.5%, 65.8%, 52.4%, 50.3%, 34.2%, 25.2%, and 12.3% towards, ampicillin/sulbactam, gentamicin, trimethoprime-sulfamethoxazole, ciprofloxacin, piperacillin-tazobactam, amikacin, amoxicillin-clavulanate, and levofloxacin respectively. Molecular characterization by PCR was employed using specific primers for three different ESBLs (TEM, SHV, and CTX-M). Results obtained revealed that SHV-type ESBLs were the most common ESBL occurring in 87% of the isolates with phenotypic evidence of ESBLs production. While those for TEM-type and CTX-M-type were 60% and 58% respectively.