Molecular Identification of Escherichia coli Pathotypes and their Antimicrobial Resistance Profiles in Stool Samples Isolated from Patients in the West of Iran.

BACKGROUND: Gastroenteritis refers to an infection in the stomach and small intestine that may be caused by bacteria, viruses, and other pathogenic agents. Most strains of Escherichia coli (E. coli) in the gastrointestinal system have shared a symbiotic relationship with humans, but some serotypes are pathogenic. This study aimed to identify E. coli pathotypes isolated from stool samples and determine the antibiotic resistance profiles of these pathotypes in the west of Iran. METHODS: The study was conducted on 106 samples of diarrheal feces which were sent to Imam Reza laboratory. First E. coli was detected and then the DNA was extracted. Next, the antibiotic sensitivity test was performed by the disk diffusion method. The E. coli pathotypes were qualitatively detected using the Amplisense Escherichioses-FRT PCR kit after DNA extraction from E. coli isolated in the stool sample. RESULTS: In this study, out of 106 E. coli-positive samples, pathogenic E. coli were detected in 62 samples including 5 samples (8.1%) which only contained the EPEC pathotype, 10 samples (16.1%) contained only the EAEC pathotype, and 12 samples (19.4%) had only the EHEC pathotype. ETEC and EIEC were not isolated from any of the samples. The sensitivity to Meropenem (97%) and Gentamicin (96.2%) showed the highest frequency among the samples. The highest level of resistance was related to Amoxicillin (93.4%) and Ampicillin (78%). CONCLUSIONS: The epidemiological results show that the predominant pathotype among all isolates is EHEC and most antibiotic resistances were related to Amoxicillin and Ampicillin. Finally, a comprehensive molecular diagnosis of E. coli pathotypes, investigation of their incidence, and antibiogram profiles will help to determine better diagnostic and therapeutic measures for managing diarrheal diseases.

Modified gold nanoparticle colorimetric probe-based biosensor for direct and rapid detection of Mycobacterium tuberculosis in sputum specimens.

The incidence of Mycobacterium tuberculosis (MTB) is increasing due to lack of appropriate diagnostic and therapeutic methods. Therefore, early and accurate detection of this bacteria plays a significant role in controlling tuberculosis. This study aimed to design, develop, and implement a direct and rapid detection method of MTB using modified gold nanoparticle (AuNP) colorimetric probe-based biosensor in sputum specimens. Spherical AuNPs were synthesized by the citrate reduction method and were functionalized using thiol-modified oligonucleotides (AuNP-biosensor). AuNP-biosensor and IS6110 PCR were compared to the gold standard in terms of analytical and clinical sensitivity and specificity, positive predictive value (PPV), negative predictive value (NPV), diagnostic odds ratio (DOR), and accuracy in 52 clinical specimens. Gold standard was defined as a positive result in concentrated sputum smear microscopy (SSM), culture, or Xpert MTB/RIF.The AuNP-biosensor had 100% sensitivity and specificity for detection of total sputum DNA in less than 15 min with ready-to-use AuNP-biosensor. PPV, NPV, DOR and accuracy of this method were 100%, 100%, 2325 and 100%, respectively. Considering the promising results of the diagnostic value indices of the AuNP-biosensor, the designed method is an affordable, rapid, reliable, and cost-beneficial way for direct detection of MTB in sputum specimens.

Multilocus Sequence Typing for Molecular Epidemiology of Stenotrophomonas maltophilia Clinical and Environmental Isolates from a Tertiary Hospital in West of Iran

Background: Stenotrophomonas maltophilia is an opportunistic bacterium, contributing to different hospital-acquired infections and can be acquired from different hospital setting sources. Epidemiological study of S. maltophilia in the hospital also demonstrates the intrahospital distribution of certain strains of bacteria in healthcare facilities. The aim of the current study was to identify the molecular epidemiology of S. maltophilia isolates from clinical and environmental sources within a hospital. Methods: A total of 400 samples (clinical and environmental) were collected from the different settings of hospital. Following the standard biochemical testing and 23S rRNA genotyping, the molecular typing of S. maltophilia isolates was determined using the multilocus sequence typing (MLST) technique. Also, the frequencies of zot and entF virulence genes among S. maltophilia isolates were examined by PCR technique. Results: Based on the biochemical testes and PCRs targeting 23S rRNA gene, 22 S. maltophilia isolates were identified. The MLST analysis demonstrated that these isolates were assigned to 14 ST, and 6 out of 14 STs were common among clinical and environmental samples. All 22 isolates were identified in the PubMLST database. The PCR screening demonstrated that none of 22 S. maltophilia isolates had zot virulence gene, while the entF gene with the 59% frequency was observed in 13 out of 22 isolates. Among these 13 isolates, 6 STs were common in clinical and environmental isolates. Conclusion: Our study showed the clonal relatedness between clinical and environmental sources of the S. maltophilia isolates in a hospital. Further studies are required to understand the epidemic situation of this pathogen in the clinic and the environment.

Molecular typing of multi-drug resistant Acinetobacter baumannii isolates from clinical and environmental specimens in three Iranian hospitals by pulsed field gel electrophoresis.

BACKGROUND: Multi-drug resistant (MDR) Acinetobacter baumannii is one of the most important causes of nosocomial infections. The purpose of this study was to identify antibiotic resistance patterns, biofilm formation and the clonal relationship of clinical and environmental isolates of A. baumannii by Pulsed Field Gel Electrophoresis method. Forty-three clinical and 26 environmental isolates of the MDR A. baumannii were collected and recognized via API 20NE. Antibiotic resistance of the isolates was assessed by the disk diffusion method, and the biofilm formation test was done by the microtiter plate method. Pulsed Field Gel Electrophoresis (PFGE) was used to assess the genomic features of the bacterial isolates. RESULTS: The resistance rate of clinical and environmental isolates against antibiotics were from 95 to 100%. The difference in antibiotic resistance rates between clinical and environmental isolates was not statistically significant (p > 0.05). Biofilm production capabilities revealed that 31 (44.9%), and 30 (43.5%) isolates had strong and moderate biofilm producer activity, respectively. PFGE typing exhibited eight different clusters (A, B, C, D, E, F, G, and H) with two significant clusters included A and G with 21 (30.4%) and 16 (23.2%) members respectively, which comprises up to 53.6% of all isolates. There was no relationship between biofilm formation and antibiotic resistance patterns with PFGE pulsotypes. CONCLUSIONS: The results show that there is a close relationship between environmental and clinical isolates of A. baumannii. Cross-contamination is also very important that occurs through daily clinical activities between environmental and clinical isolates. Therefore, in order to reduce the clonal contamination of MDR A. baumannii environmental and clinical isolates, it is necessary to use strict infection control strategies.

Code and data on the processing of the pulsed-field gel electrophoresis images: A matlab script.

Here a matlab script was presented for lane tracking and band detection on the pulsed field gel electrophoresis (PFGE) images. It can also be used as a software tool for automatic analysis of PFGE images. The data consist of several MATLAB codes which collectively have the task of lane tracking, band detecting and pattern recognition on the PFGE images. The lane tracking stage is semi-automatic and the band detection stage is fully automatic. Finally, the pattern of lanes that includes number of, location, width and light intensity level of bands was obtained.

Molecular characterization of clinical and environmental Pseudomonas aeruginosa isolated in a burn center.

In burn centers, Pseudomonas aeruginosa acts as a major cause of nosocomial infections. Therefore, this study aimed to characterize molecularly P. aeruginosa isolates collected from environmental samples and burn patients. A total of 78 strains (including 58 clinical and 20 environmental isolates) of the P. aeruginosa were collected from Beasat hospital of Hamadan, west of Iran, and was identified using API 20NE. The disk diffusion method according to the CLSI was applied for determination of the antimicrobial resistance. Moreover, the microtiter plate test was used for the quantification of Biofilm formation. The genomic features of the isolated strains was evaluated using Pulsed Field Gel Electrophoresis (PFGE). We found that 94.8% of clinical and 80% environmental isolates were capable of forming biofilm. The rate of MDR in clinical and environmental isolates was 51.7% and 40%, respectively. A significant relationship was observed between biofilm formation capability and multiple drug resistance (p < 0.05). PFGE typing showed 11 different clusters with two major clusters A with 30 (38.5%) and B with 14 (17.9%) members, containing up to 56.4% of all isolates. There was no relationship between biofilm formation ability and antibiotic resistance patterns with PFGE patterns. According to the results, the clonal spread of environmental P. aeruginosa isolates is associated with clinical isolates, and both environmental and clinical isolates are attributed to a high prevalence of the antibiotic resistance and biofilm formation ability. This study highlighted that the prevention programs should be implemented in the hospital environment to control the spread of P. aeruginosa in burn units.

Genomic diversity and antimicrobial susceptibility profiling of nasal carriage Staphylococcus aureus isolated from pediatric ward in Western Iran.

Nasal carriage of Staphylococcus aureus (S. aureus) probably causes the transmission of infection between individuals in hospital and community. This study aimed to evaluate the molecular epidemiology and antibiotic resistance pattern of nasal carriage S. aureus in pediatric ward patients and personnel. A total of 122 Nasal samples were taken from 28 personnel and 94 hospitalized patients in the pediatric ward. Minimum Inhibitory Concentration (MIC) to vancomycin and cefoxitin was determined by Agar dilution method strips. All S. aureus isolates were analyzed by pulsed-field gel electrophoresis (PFGE). A total of 41 S. aureus were isolated from the patients. 16 isolates (39.09%) were hospital-associated S. aureus (HA-SA) and 25 (60.97%) were community-associated S. aureus (CA-SA); also, 13 S. aureus isolates were obtained from the personnel. Based on MIC results, all of S. aureus isolates were susceptible to vancomycin, and in 41 patient isolates, 13 isolates (31.7%) were resistant to cefoxitin (MRSA). Of 13 S. aureus isolates of the personnel, 3 (23%) isolates were MRSA. Totally 11 common clones and 13 single clones were obtained. In conclusion the prevalence of CA-SA in the ward was higher than that of HA-SA. In the strains obtained from a hospital ward, there was a high epidemiology, genotypic diversity in the studied ward. However, horizontal transfer of S. aureus was observed between patients and between personnel and patients, which indicated the risk of transmission of resistant strains in the hospital wards.

Prevalence of nontuberculous mycobacteria and high efficacy of d-cycloserine and its synergistic effect with clarithromycin against Mycobacterium fortuitum and Mycobacterium abscessus.

BACKGROUND: The prevalence of pulmonary disease caused by nontuberculous mycobacteria (NTM) is reportedly on the rise in the world. Some of the species are resistant to various antibiotics; hence, limited treatment options are available. The aims of this study were to investigate the prevalence of NTM and to determine the effect of d-cycloserine against Mycobacterium fortuitum and Mycobacterium abscessus isolated from clinical specimens to find out the synergistic effect of d-cycloserine and clarithromycin. METHODS: A total of 95 nonduplicate pulmonary isolates of NTM were collected from three major Regional Tuberculosis (TB) Centers. NTM isolates were identified by conventional tests and PCR sequence analysis of the rpoB gene. PCR sequencing of erm-41 was performed for detecting the inducible resistance to macrolides. In vitro susceptibilities and activities of d-cycloserine-clarithromycin combinations were accessed using the broth microdilution method. RESULTS: Among 714-positive acid-fast bacilli from TB-suspected cases, 95 isolates were identified as NTM (13.3%). The prevalence of identified isolates was as follows: M. fortuitum 46 (48.4%), Mycobacterium simiae 16 (16.8%), Mycobacterium kansasii 15 (15.7%), M. abscessus 7 (7.3%), Mycobacterium thermoresistibile 4 (4.2%), Mycobacterium elephantis 3 (3.2%), Mycobacterium porcinum 2 (2.1%), and Mycobacterium chimaera 2 (2.1%). In addition, rpoB sequence analysis could identify all NTM isolates. The effect of d-cycloserine was better than that of clarithromycin. The synergistic effect of d-cycloserine with clarithromycin was observed for six (100%) and five (71.5%) strains of M. fortuitum and M. abscessus, respectively. CONCLUSION: In the present study, we demonstrated a wide range of NTM in processed samples from different provinces of Iran. Our observations indicated that d-cycloserine was very active against M. abscessus and M. fortuitum; hence, d-cycloserine, either alone or in combination with clarithromycin, may be promising for the treatment of M. abscessus- and M. fortuitum-associated diseases.

Genotyping and distribution of putative virulence factors and antibiotic resistance genes of Acinetobacter baumannii strains isolated from raw meat.

Background: Acinetobacter baumannii strains with multiple antimicrobial resistance are primarily known as opportunistic nosocomial bacteria but they may also be regarded as emerging bacterial contaminants of food samples of animal origin. Here we aimed to study the molecular characteristics of the A. baumanni strains isolated from raw meat samples. Methods: A total of 22 A. baumanni strains were isolated from 126 animal meat samples and were genotyped by ERIC-PCR method and by PCR detection of their virulence and antimicrobial resistance determinants. A. baumannii strains with 80% and more similarities were considered as one cluster. Results: Sixteen different genetic clusters were found amongst the 22 A. baumanni strains. Of the 22 strains, 12 (54.54%) had similar genetic cluster. A. baumannii strains exhibited the highest percentage of resistance against tetracycline (90.90%), trimethoprim (59.09%), cotrimoxazole (54.54%) and gentamicin (50.00%). TetA (81.81%), tetB (72.72%), dfrA1 (63.63%), aac(3)-IV (63.63%), sul1 (63.63%) and aadA1 (45.45%) were the most commonly detected antibiotic resistance genes. FimH (81.81%), afa/draBC (63.63%), csgA (63.63%), cnf1 (59.09%), cnf2 (54.54%) and iutA (50.00%) were the most commonly detected virulence factors. A. baumannii strains isolated from the chicken meat samples had the highest similarities in the genetic cluster. Conclusions: A. baumannii strains with similar genetic cluster (ERIC-Type) had the same prevalence of antibiotic resistance, antibiotic resistance genes and virulence factors. Genetic cluster of the A. baumannii strains is the main factor affected the similarities in the genotypic and phenotypic properties of the A. baumannii strains.

Clonal Lineage Diversity, Antibiotic Resistance, and Virulence Determinants Among Methicillin-Resistant and Methicillin-Susceptible Staphylococcus aureus Isolated from Nurses at a Teaching Hospital in Ilam, Iran: Successful Nares Decolonization by Mupirocin.

BACKGROUND: Staphylococcus aureus is known to be responsible for nosocomial infections, and the typing method was useful in managing the reservoir of bacteria. The main aim of this study was to determine the prevalence of S. aureus in the nares and hands of nurses working in Imam Khomeini hospital, Ilam, Iran, as well as to determine the clonal relatedness, antimicrobial susceptibility profiles, different virulence, and resistance determinants among these isolates. The evolution of mupirocin activity in the eradication of methicillin-resistant S. aureus (MRSA) and methicillin-susceptible S. aureus (MSSA) colonization in the nares of the healthcare workers in Ilam, Iran, was also determined in this study. MATERIALS AND METHODS: In this cross-sectional study, 80 nurses, auxiliary nurses, and service workers from Imam Khomeini Hospital were enrolled. MRSA, antibiotic susceptibility, and virulence determinants were evaluated. Then, the isolates were subjected to pulsed field gel electrophoresis (PFGE) and Staphylococcal cassette chromosome mec typing. RESULTS: Our results demonstrated that 23% of isolates were MRSA. PFGE results demonstrated that pulsotypes A (3 out of 30; 10%) and J (3 out of 30; 10%), pulsotypes E (2 out of 30; 6.7%), M (2 out of 30; 6.7%), P(2 out of 30; 6.7%), and V (2 out of 30; 6.7%) were the most predominant pulsotypes, respectively. CONCLUSION: We cannot give conclusive suggestions about the correlation between nasal carriage and infections, but we suggest the monitoring of all healthcare workers annually, decontamination of their noses by using mupirocin and other antistaphylococcal agents, and also the washing of their hands at least every 2 h.

Molecular identification and distribution of non-tuberculous mycobacteria isolated from clinical specimens by PCR-sequencing method in West of Iran.

BACKGROUND AND AIMS: Infections by non-tuberculous mycobacteria (NTM) has rapidly increased in recent years, due to high infection rates related to the populations at risk like immunocompromised individuals, patients predisposed by prior pulmonary. The aim of this study was to investigate the presence of NTM in clinical samples and genetic diversity using 16S rRNA and rpoB sequence analysis. METHODS: A cross-sectional study was conducted on 45 diverse isolates collected from sputum in 2 years 2014-2015 using standard decontamination procedure. All mycobacterial isolates were grown on LJ medium and also conventional tests for preliminary identification of mycobacteria rely on traits and then DNA extraction. PCR was performed, and sequencing of 16S rRNA and rpoB genes was applied for NTM strains identification. RESULTS: A total of 45 isolates collected, 37 samples (83%) were evaluated as NTM. All NTM strains using molecular methods by sequencing 16S rRNA and rpoB gene were identified, by this way 12 different species have been identified which sequencing of rpoB was able to identify all species. The major species obtained were Mycobacterium simiae (22%), M. fortuitum (19%), and M. abscessus (13%). CONCLUSIONS: The results of our study showed that the patients were infected by a wide range of atypical mycobacteria. It was concluded that 16S rRNA gene sequencing coupled with rpoB marker is a high discriminatory power in identification of NTM. The presence of various species in clinical samples in Iran emphasizes the use of molecular method like sequence analysis of genes is necessary for reliable identification.

Mycobacterium tuberculosis of the Beijing Genotype in Iran and the World Health Organization Eastern Mediterranean Region: A Meta-Analysis.

OBJECTIVE: The Beijing genotype is a distinct genetic lineage of Mycobacterium tuberculosis, which is distributed worldwide, and may cause large outbreaks of multidrug resistance-tuberculosis (MDR-TB). The distribution of such strains in the Eastern Mediterranean region (EMR) is unclear, and clarifying the data is our purpose apart from the presence of Beijing TB in Iran. METHODS: We searched Published literature from CINAHL Cochrane Library, Current Contents, Database of Abstracts of Reviews of Effects (DARE), ProQuest Google Scholar PubMed, PsycINFO, Thomson Reuters, (SID), and Medical Library (MedLib) to detect relevant studies from the year 2000 to July 2015 with the following keywords: M. tuberculosis, Beijing genotype, EMR, and drug resistance. Random-effect models were used to estimate the proportion of Beijing strains in STATA 14. Heterogeneity was investigated by subgroup analysis and meta-regression. RESULTS AND CONCLUSION: The meta-prevalence of Beijing strains was 4% (CI 95% = 3-5). The prevalence was different based on types of detection techniques (spoligotyping = 4% vs. other techniques = 6%; p = 0.003) and years of study (before the year 2000 = 2% vs. after year 2000 = 4%, p = 0.004). The Beijing family was most prevalent in Iran and Pakistan. A strong relationship with drug resistance was reported in Pakistan and Iran, and an increasing trend was seen in Pakistan. Additional studies of drug-resistant TB distribution among Beijing strains in EMR countries are needed as well as a time-trend analysis of the Beijing strain infection in the region.

The relationship between embb306 and embb406 mutations and ethambutol resistant in Mycobacterium tuberculosis isolated from patiens in west of Iran.

Background: Mutations in embB gene have been reported in ethambutol (EMB) resistance Mycobacterium tuberculosis (M. tuberculosis) isolates. The aim of this study was survey on embB 306 and 406 EMB resistant M. tuberculosis isolated from patients in West of Iran (2014-2015). Methods: Fifty strains of M. tuberculosis from patients with pulmonary tuberculosis (TB) were considered. Drug susceptibility using proportional method was performed. Polymerase chain reaction (PCR) -DNA sequencing was applied for mutation in embB 306 and 406 codon detection. Data were analyzed in SPSS 16 software using descriptive statistics and Fisher's exact test (p<0.05). Results: In this study 7 (14%) M. tuberculosis isolates were resistant to EMB. 6 (85.71%) and 1 (14.28%) resistant isolates had embB 306 and 304 codon mutations, respectively. Between embB306 mutations and resistance to EMB and MDR isolates had a significant relationship (p<0.001). Conclusion: The data indicated that embB 306 mutations have effect on EMB resistant. Detection of EMB resistant and these mutations prominent for antibiotic prescription.

Genetic diversity of multidrug-resistant Mycobacterium tuberculosis strains isolated from tuberculosis patients in Iran using MIRU-VNTR technique.

Tuberculosis (TB) is considered as one of the most important infectious diseases in the world, and recent rise and spread of multidrug-resistant (MDR) Mycobacterium tuberculosis (MTB) strains, have made the matter worsened. Due to the importance of TB prevalence in Iran, this study was designed to investigate the genetic diversity among MDR strains of MTB by MIRU-VNTR typing scheme. A total of 88 drug resistant M. tuberculosis isolates belong to pulmonary TB cases were collected from several TB reference centers of Iran. Drug susceptibility testing for Isoniazid and Rifampin was performed using the agar proportion method and MDR isolates were underwent genotyping by using 12-locus- based MIRU-VNTR typing. On performing proportion method, 22 isolates were identified as MDR. By typing of MDR isolates using 12-loci MIRU-VNTR technique, high diversity were demonstrated in MDR strains and these were classified into 20 distinct MIRU-VNTR genotypes. MIRU loci 10 and 26 were the most discriminatory loci with 8 and 7 alleles respectively; while MIRU loci 2, 20, 24 and 39 were found to be the least discriminatory with 1-2 alleles each. We noticed a mixed infection in isolate 53, as this isolate comprised simultaneous two alleles in MIRU loci 40, 10, 16 and 39. In conclusion, this result represents MIRU-VNTR typing as a useful tool for studying genetic diversity of MDR-MTB in regional settings, and will help the health sectors to construct a preventive program for MDR-TB. Additionally, it can detect mixed infection which can facilitate management of treatment.

Frequency of Class 1 Integron and Genetic Diversity of Acinetobacter baumannii Isolated from Medical Centers in Kermanshah.

BACKGROUND AND OBJECTIVE: Acinetobacter baumannii has emerged as an important pathogen in hospital and environment that can acquire transport element and antibiotic-resistant genes. The aim of this study was to determine the resistances to different antibiotics, frequency of Class 1 integron in A. baumannii and then molecular typing for A. baumannii isolated from Intensive Care Unit (ICU). MATERIALS AND METHODS: A total of 100 isolates of A. baumannii were collected from patients admitted to hospitals in Kermanshah from April 2014 to September 2015. The isolates were identified using biochemical test. Antimicrobial susceptibility test for 20 antibiotics was determined by Kirby-Bauer antibiotic testing (or disc diffusion). The prevalence rate of class integrons among the isolates was determined using polymerase chain reaction and finally 80 isolates of A. baumannii obtained from the Intensive Care Unit were selected for molecular typing by pulsed-field gel electrophoresis (PFGE). RESULTS: The maximum drug resistance was observed against cefotaxime, ceftriaxone, mezlocillin, imipenem, and ceftazidime and piperacillin. Twenty-nine isolates were multidrug resistant (MDR); about 21 isolates were extensively-drug resistant and none were pandrug resistance and 42 isolates (42%) contained Class 1 integrons. The results did not show a significant correlation between the presence of Class 1 integrons and incidence of MDR A. baumannii. Five clusters were obtained by PFGE. CONCLUSION: This study did not show a significant correlation between the presence of Class 1 integrons and incidence of MDR A. baumannii. By PFGE analysis, the high level of similarity between some pulsotypes in A. baumannii strains showed genetic correlation between them.

Molecular Characterization of Multidrug Resistant Strains of Acinetobacter baumannii Isolated from Intensive Care Units in West of Iran.

INTRODUCTION: According to the results of various studies using phenotypic methods, the prevalence of Multidrug Resistant (MDR) Acinetobacter baumannii (A. baumannii) isolates has been increasing worldwide. Pulsed-Field Gel Electrophoresis (PFGE) technique is known as the gold standard method to determine clonal characterization of bacterial species, especially A. baumannii. AIM: To determine the clonal relatedness and investigate the prevalence of integron classes 1 and 2 and genes encoding OXA-23 and 24 in A.baumanii isolates. MATERIALS AND METHODS: A cross-sectional study was conducted from November 2011 to January 2013. A total of 140 A.baumannii isolates collected from three hospitals of Kermanshah were considered out of which 75 ICU isolates were included in this study. Antibiotics susceptibility test was done by disk diffusion method. Polymerase Chain Reaction (PCR) was performed in order to detect class 1 and 2 integrons and blaOXA-23-like, blaOXA-24-like genes. Isolates identified as MDR from a total of 75 Intensive Care Units (ICU) strains were subjected to genotyping for clonal relatedness. RESULTS: A total of 37 isolates among 75 ICU isolates were identified as MDR. The maximum drug resistance was observed against ceftriaxone, mezlocycline, cefotaxime, piperacilin, ciprofloxacin and imipenem. Frequency of Class 1 and Class 2 Integrons, blaOXA-23-like and blaOXA-24-like genes were 33(44%), 27(36%), 60(80%) and 14(18.6%) respectively. Four clusters with high level of similarity were obtained showing homogeneity among MDR isolates. CONCLUSION: Significant correlation between presence of integrons and resistance to different classes of antibiotic was observed in this study. Monitoring of drug resistance using gene integrase PCR and blaOXA gene by cluster analysis is very important to plan specific infection control measures due to MDR A. baumannii.

Verification of Frequency in Species of Nontuberculous Mycobacteria in Kermanshah Drinking Water Supplies Using the PCR-Sequencing Method.

BACKGROUND: Nontuberculous mycobacteria are habitants of environment, especially in aquatic systems. Some of them cause problems in immunodeficient patients. Over the last decade, 16S rRNA gene sequencing was established in 45 novel species of nontuberculous mycobacteria. Experiences revealed that this method underestimates the diversity, but does not distinguish between some of mycobacterium subsp. To recognize emerging rapidly growing mycobacteria and identify their subsp, rpoB gene sequencing has been developed. OBJECTIVES: To better understand the transmission of nontuberculous mycobacterial species from drinking water and preventing the spread of illness with these bacteria, the aim of this study was to detect the presence of bacteria by PCR-sequencing techniques. MATERIALS AND METHODS: Drinking water samples were collected from different areas of Kermanshah city in west of IRAN. After decontamination with cetylpyridinium chloride, samples were filtered with 0.45-micron filters, the filter transferred directly on growth medium waiting to appear in colonies, then DNA extraction and PCR were performed, and products were sent to sequencing. RESULTS: We found 35/110 (32%) nontuberculous mycobacterial species in drinking water samples, isolates included Mycobacterium goodii, Mycobacterium aurum, and Mycobacterium gastri with the most abundance (11.5%), followed by Mycobacterium smegmatis, Mycobacterium porcinum, Mycobacterium peregrinum, Mycobacterium mucogenicum, and Mycobacterium chelonae (8%). CONCLUSIONS: In this study, we recognized the evidence of contamination by nontuberculous mycobacteria in corroded water pipes. As a result of the high prevalence of these bacteria in drinking water in Kermanshah, this is important evidence of transmission through drinking water. This finding can also help public health policy makers control these isolates in drinking water supplies in Kermanshah.

Beijing strains of Mycobacterium tuberculosis in smear-positive tuberculosis patients in North-West and West of Iran.

BACKGROUND: Tuberculosis (TB) is the leading cause of morbidity and mortality among chronic infectious diseases. The goal of this cross-sectional study (2012-2014) was to examine the prevalence of Mycobacterium TB (MTB) Beijing strains in regions near the Iranian border and to identify any epidemiological links. MATERIALS AND METHODS: To this end, MTB isolates were harvested, from 64 HIV-negative, pulmonary smear-positive TB patients from the Iranian border provinces of East Azerbaijan (North-West), Kurdistan (West), and Kermanshah (West) (2012-2014). Isolates were subjected to restriction fragment length polymorphism (RFLP) analysis, using the insertion sequence IS6110 as a probe (IS6110 RFLP), and drug susceptibility testing by the proportion method. We gathered demographic and clinical data using a questionnaire and reviewing patient records. Results were analyzed with Gel Compare II 6.6 and SPSS-18. RESULTS: The mean age of the patients was 54.4 years and 46.9% were male. The prevalence of Beijing strains among the isolates was 9.4% (17.6% in the Western provinces and 0% in East Azerbaijan). There was a statistically significant relationship between the Beijing strains and drug resistance and also between these strains, and the recurrence of TB in patients that had previously received treatment (P = 0.02 and P = 0.04, respectively). CONCLUSIONS: Finally, the prevalence of Beijing strains in Western Iran was greater than expected. Our results therefore indicate that regional and cross-border tracing may be necessary to control spread of this organism.

Mycobacterium tuberculosis Beijing Genotype in Western Iran: Distribution and Drug Resistance.

INTRODUCTION: Mycobacterium tuberculosis Beijing genotype is gaining importance all over the world because this genotype is highly prevalent in several areas and is also frequently associated with drug resistance. AIM: To identify and determine the frequency of Beijing genotype and mix infection with Beijing and non-Beijing in west of Iran and analyse the association between Beijing genotype and drug resistance. MATERIALS AND METHODS: This cross-sectional study was conducted on 146 Tuberculosis (TB) samples collected at the TB reference laboratory in Kermanshah west of Iran from January 2014 to February 2015, Mycobacterium tuberculosis isolates from sputum samples, detected by microcopy, biochemical tests and solid culture were included and then the confirmed samples with Cepheid Xpert MTB/RIF assay were subjected to drug susceptibility tests for rifampicin, isoniazid, ethambutol using proportional method. The prevalence rate of Beijing and non-Beijing genotype was determined by Multiplex- Polymerase Chain Reaction (PCR). RESULT: A total of 15/146 (10%) isolates were diagnosed as Beijing genotypes and the remaining 131/146(90%) isolates were non-Beijing genotypes by Multiplex PCR method. Among the 15 Beijing cases, 14 samples have shown mix infection indicating the presence of both Beijing and non-Beijing strains in samples. Three isolates from all cases were drug resistant. Interestingly all drug resistance isolates were from Beijing genotype which shows strong association between drug resistance and Beijing genotype. Also this genotype was more prevalent in younger age-group people (p=0.035). CONCLUSION: Frequency of Beijing genotype in west of Iran is more than other sites of Iran but less than Asia. According to our result, mix infections with Beijing and non-Beijing, had the most prevalence therefore we should be concerned more about mix infections. Multiplex-PCR method is feasible, trustworthy and can distinguish mix infections. It is suggested to perform spoligotyping in addition to multiplex PCR method to discriminate mix infections.

Patterns of Drug Resistance Among Tuberculosis Patients in West and Northwestern Iran.

BACKGROUND: Tuberculosis (TB) is the leading cause of morbidity and mortality among chronic infectious diseases. OBJECTIVE: The goal of this cross-sectional study (2011-2013;2013) was to examine the patterns of TB drug resistance among HIV-negative pulmonary TB patients in regions near the Iranian border. METHOD: To this end, MTB isolates were harvested from 300 HIV-negative, pulmonary smear-positive TB patients from the northwest and west Iranian border provinces. Isolates were subjected to first and second-line drug susceptibility testing by the 1% proportion method. Demographic and clinical data were provided using a questionnaire and information from patient records. Results were analyzed using SPSS-18. RESULTS: The mean age of the patients was 52.03 years and 54.3% were male. The prevalence of resistance to any TB drug was 13.6% (38 cases). Eleven percent of the new treatment TB group (28 patients) and 40.7% of the retreatment TB group (11 patients) were resistant to all TB drugs. Twelve (4.3%) patients had multidrug-resistant tuberculosis (MDR-TB) (2.38% in the new TB treatment group and 23.1% in the retreatment group). One patient had extensively drug-resistant tuberculosis (XDR-TB). There was a statistically significant relationship between TB drug resistance and smoking (p=0.02) and a history of migration from village to city (p=0.04), also between TB drug resistance and recurrence of TB in patients that had previously received treatment (p<0.001). CONCLUSION: Knowledge of drug resistance patterns for new and previously treated cases is critical for effective control of MDR-TB in different regions of the country. The burden of MDR-TB in retreatment cases was high. Previous TB treatment was one of the most important mokers and those who had a history of rural to urban migration were at high risk for the occurrence of TB drug resistance.