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Background: Bacterial resistance to antibiotics limits treatment options, increases morbidity and mortality, and raises the risk of antibiotic-associated adverse events. Antibacterial resistance emerges rapidly following an increase in the consumption of antibiotics against infectious diseases. The spread of ESBL producing strains has a limiting factor based on antibiotic function for the treatment of infections particularly caused by Acinetobacter baumannii ( A. baumannii )., Objective: This study was conducted to evaluate the prevalence of antimicrobial resistance and distribution of bla TEM , bla CTX , and bla SHV genes among A. baumannii strains isolated from clinical samples at a major hospital in Teheran, Iran., Methods: A. baumannii strains were isolated and identified using standard microbiological methods. The disc diffusion and combined discs methods were used for testing antimicrobial susceptibility and to identify the strains producing Extended-Spectrum Beta-Lactamases (ESBL), respectively. DNA extraction was done by boiling method. Finally, the frequency of resistant genes including bla TEM , bla CTX , and bla SHV in ESBL producing isolates was studied by PCR., Results: Gender distribution in this study was 53 (53%) samples for men and 47 (47%) for women. Totally, one hundred A. baumannii strains were isolated. More than 93% of the isolates were multi drug resistant. The highest to lowest antibiotic resistance was observed against amoxicillin/clavulanic acid (98%), ceftriaxone (96%), cefotaxime (94%), and ceftazidime (93%), respectively. The frequency of positive phenotypic test of ESBL was 19% and 16% for CAZ-C and CTX-C, respectively. The frequency of bla TEM , bla CTX , and bla SHV genes was 52.1, 43.4, and 21.7, respectively., Conclusion: A. baumannii isolates exhibited an extremely worrying level of antibiotic resistance, and a high percentage of the isolates showed MDR in this study. This is a serious warning because ESBLs are a major threat to the effectiveness of antibiotics that are currently available for medical uses. The frequency of genes encoded ESBL isolates of A. baumannii may be due to overuse and misuse of antibiotics.

Background: Escherichia coli is one of the most important bacterial agents to cause urinary tract infections. Inappropriate and unnecessary administration of antibiotics has led to an increase in the appearance of multidrug-resistant E. coli isolates, limiting treatment options. The increase in a number of resistant strains of bacteria is a major concern of health authorities worldwide., Objective: The purpose of this study was to determine the presence of the qnr genes among E. coli isolated from UTIs of patients in Baqiyatallah hospital in Tehran province, Iran., Method: Clinical urine samples of patients with suspected urinary tract infection were collected by standard methods in sterile disposable containers. After analysis of urine, microscopic observations and culture analysis, the bacterial genome was extracted by boiling method. PCR for detection of qnr genes including qnrA , qnrB and qnrS was done by specific primers, then PCR products were run using gel electrophoresis and visualized by gel documentation system., Results: In the present study among the 95 isolates, 60 strains were resistant to nalidixic acid. PCR showed that 92 strains were positive for qnrS . The qnrA and qnrB genes were not found among the clinical isolates., Conclusion: Our finding indicates a high level of resistance against nalidixic acid among E. coli isolates recovered from the patients with UTI. Also, the high frequency of qnrS imposes the importance of survey of molecular and genetic analysis of mechanisms of quinolone resistance in E. coli strains.

Introduction: CRISPR (Clustered Regularly Interspaced Short Palindromic Repeats) loci as novel and applicable regions in prokaryotic genomes have gained great attraction in the post genomics era., Methods: These unique regions are diverse in number and sequence composition in different pathogenic bacteria and thereby can be a suitable candidate for molecular epidemiology and genotyping studies. Results:Furthermore, the arrayed structure of CRISPR loci (several unique repeats spaced with the variable sequence) and associated cas genes act as an active prokaryotic immune system against viral replication and conjugative elements. This property can be used as a tool for RNA editing in bioengineering studies., Conclusion: The aim of this review was to survey some details about the history, nature, and potential applications of CRISPR arrays in both genetic engineering and bacterial genotyping studies.

Background: A rapid, accurate, flexible and reliable diagnostic method may significantly decrease the costs of diagnosis and treatment. Designing an appropriate microarray chip reduces noises and probable biases in the final result., Objective: The aim of this study was to design and construct a DNA Microarray Chip for a rapid detection and identification of 10 important bacterial agents., Method: In the present survey, 10 unique genomic regions relating to 10 pathogenic bacterial agents including Escherichia coli (E.coli), Shigella boydii, Sh.dysenteriae, Sh.flexneri, Sh.sonnei, Salmonella typhi, S.typhimurium, Brucella sp., Legionella pneumophila, and Vibrio cholera were selected for designing specific long oligo microarray probes. For this reason, the in-silico operations including utilization of the NCBI RefSeq database, Servers of PanSeq and Gview, AlleleID 7.7 and Oligo Analyzer 3.1 was done. On the other hand, the in-vitro part of the study comprised stages of robotic microarray chip probe spotting, bacterial DNAs extraction and DNA labeling, hybridization and microarray chip scanning. In wet lab section, different tools and apparatus such as Nexterion® Slide E, Qarray mini spotter, NimbleGen kit, TrayMix TM S4, and Innoscan 710 were used., Results: A DNA microarray chip including 10 long oligo microarray probes was designed and constructed for detection and identification of 10 pathogenic bacteria., Conclusion: The DNA microarray chip was capable to identify all 10 bacterial agents tested simultaneously. The presence of a professional bioinformatician as a probe designer is needed to design appropriate multifunctional microarray probes to increase the accuracy of the outcomes.

Background: Antimicrobial resistance is an important factor threatening human health. It is widely accepted that antibiotic resistant bacteria such as Escherichia coli ( E. coli) released from humans and animals into the water sources, can introduce their resistance genes into the natural bacterial community., Objective: The aim of this study was to investigate the prevalence of bla TEM , bla CTX , bla SHV , bla OXA and bla VEB associated-antibiotic resistance among E. coli bacteria isolated from different water resources in Iran., Methods: The study contained all E. coli strains segregated from different surface water sources. The Kirby-Bauer method and combined discs method was determined in this study for testing antimicrobial susceptibility and strains that produced Extended-Spectrum Beta Lactamases (ESBL), respectively. DNA extraction kit was applied for genomic and plasmid DNA derivation. Finally the frequency of resistant genes including bla TEM , bla CTX , bla SHV , bla OXA and bla VEB in ESBL producing isolates were studied by PCR., Results: One hundred E. coli strains were isolated and entered in the study. The highest antibiotic resistance was observed on clindamycin (96%). Moreover, 38.5% isolates were ESBL producers. The frequency of different ESBLs genes were 37%, 27%, 27%, and 25% for bla TEM , bla CTX , bla SHV , and bla OXA , respectively. The bla VEB wasn't found in any isolates., Conclusion: The study revealed a high prevalence of CTX-M, TEM, SHV and OXA genes among E. coli strains in surface water resources. In conclusion, these results raised a concern regarding the presence and distribution of these threatening factors in surface water sources and its subsequent outcomes.

Introduction: Salmonella is known as one of the most important causes of gastrointestinal disease in the world. Quinolones and fluoroquinolones are used successfully in the treatment of salmonellosis particularly for infections that have become resistant to several antibiotics. But non-susceptible isolates to quinolones have been reported in several countries. The data are limited about the prevalence of quinolone-resistant isolates in our country. Therefore, this study investigated the plasmid-mediated quinolone resistance genes in Salmonella enterica isolated in Children's Medical Center in Tehran during 2014-2015., Methods and Materials: Salmonella isolates were isolated and identified using standard microbiological methods. Antibiotic susceptibility testing and screening of Salmonella strains resistant to quinolones were performed according to the CLSI guidelines. The molecular investigation was done using specific primers for detection of qnr genes including: qnr A, qnr B and qnr S, by polymerase chain reaction., Results: Overall, 92 (66.6%) strains were resistant to nalidixic acid. None of the strains showed resistance to ciprofloxacin. Out of the 92 nalidixic acid resistant strains, 52 (56.52%) harbored qnr S genes, 15 strains (16.30%) had both qnr A and qnr S genes. Two (1.1%) isolates were positive for qnr B gene. Twenty four (26.08%) nalidixic acid resistant isolates did not have any qnr qens., Conclusion: The results of this study show high prevalence of resistance to nalidixic and qnr genes in Salmonella isolates. Plasmid nature of this type of resistance poses an increased risk of dissemination of quinolone resistance between Salmonella and non- Salmonella isolates circulating in hospitals environments.

Background: Francisella tularensis ( F. tularensis ) is the etiological microorganism for tularemia. There are different forms of tularemia such as respiratory tularemia. Respiratory tularemia is the most severe form of tularemia with a high rate of mortality; if not treated. Therefore, traditional microbiological tools and Polymerase Chain Reaction (PCR) are not useful for a rapid, reliable, accurate, sensitive and specific diagnosis. But, DNA microarray technology does. DNA microarray technology needs to appropriate microarray probe designing., Objective: The main goal of this original article was to design suitable long oligo microarray probes for detection and identification of F. tularensis ., Method: For performing this research, the complete genomes of F. tularensis subsp. tularensis FSC198, F. tularensis subsp. holarctica LVS, F. tularensis subsp. mediasiatica , F. tularensis subsp. novicida ( F. novicida U112), and F. philomiragia subsp. philomiragia ATCC 25017 were studied via NCBI BLAST tool, GView and PanSeq Servers and finally the microarray probes were produced and processed via AlleleID 7.7 software and Oligoanalyzer tool, respectively., Results: In this in silico investigation, a number of long oligo microarray probes were designed for detecting and identifying F. tularensis . Among these probes, 15 probes were recognized as the best candidates for microarray chip designing., Conclusion: Calibrated microarray probes reduce the biasis of DNA microarray technology as an advanced, rapid, accurate and cost-effective molecular diagnostic tool with high specificity and sensitivity. Professional microarray probe designing provides us with much more facility and flexibility regarding preparation of a microarray diagnostic chip.

Background and Objectives: Salmonella is an important food-borne pathogen responsible for disease in humans and animals. The aim of this study was to investigate the genetic relationship among third generation cephalosporin-resistant Salmonella enterica strains by Enterobacterial Repetitive Intergenic Consensus (ERIC)-PCR., Methods: The study included all Salmonella isolates obtained from clinical cases in a pediatric hospital in Tehran, Iran during 2006 to 2009. Antimicrobial susceptibility testing was performed according to the Clinical and Laboratory Standards Institute. The genetic relationship between third generation cephalosporins-resistant Salmonella enterica strains was determined using ERIC-PCR., Results: Of 136 Salmonella enterica isolates recovered from pediatric patients, six isolates including four Salmonella enterica serotype Infantis and two Salmonella enterica serotype Enteritidis showed an extended-spectrum cephalosporins resistant phenotype. ERIC-PCR differentiated Salmonella enterica serotypes Infantis and Enteritidis into 2 distinct clusters arbitrarily named as E1 and E2. Profile E1 was found in two Salmonella enterica serotype Enteritidis isolates, and profile E2 was found in four Salmonella enterica serotype Infantis isolates., Conclusion: Extended-spectrum cephalosporins resistant Salmonella could be attributed to a few predominant serotypes including Enteritidis and Infantis in this study. Genetic analysis using ERIC-PCR showed that closely related clones are responsible for the occurrence of extended-spectrum cephalosporins resistant Salmonella infection in Tehran.

Aims of this study were to investigate on antibiotic resistance and molecular epidemiology of K.pneumoniae producing ESBLs isolates of respiratory tract infections in some major hospitals in Iran. K.pneumonaie were obtained of patients with RTI. K. pneumoniae producing ESBLs detected by screening, confirming and PCR methods. During the 12-month period, a total of one hundred and thirteen of K.pneumoniae were found from RTI in three cities in different region of Iran which Sixty seven strains (59.2%) were ESBLs producer. In Ilam hospitals, seventeen strains (43.6%), in Milad hospital, thirty-seven strains (74%) and in Emam Reza hospital, thirteen strains (54.2%) were ESBLs producer. The findings showed that among sixty-seven K.pneumoniae producing ESBLs, Sixty-three strains (94%) were positive for blaSHV, eleven strains (16.4%) contained blaTEM and sixteen strains (23.9%) harbored blaCTX-M. Imipenem was found as an effectiveness antibiotic. In the current study, Majority of the ESBLs production had occurred in Milad hospital in Tehran (74%). In conclusion, spreading ESBL-producing strains is a concern, as it causes limitations to the antimicrobial agents for optimal treatment of patients.