Determining the prevalence of SCCmec polymorphism, virulence and antibiotic resistance genes among methicillin-resistant Staphylococcus aureus (MRSA) isolates collected from selected hospitals in west of Iran.

Methicillin-resistant Staphylococcus aureus (MRSA) is one of the most important pathogens worldwide and compared to other staphylococcal species that are associated with higher mortality rate. A total of 500 Staphylococcus spp. was collected from selected hospitals in Ilam, Kermanshah, Khorram Abad and Hamadan cities and, via phenotypic and genotypic methods, was assessed to find MRSA. The presence or absence of prevalent antibiotic resistance genes and virulence genes was evaluated among MRSA isolates, using polymerase chain reaction (PCR) method, and then the SCCmec typing of these isolates was assayed by multiplex PCR. A total of 372 (74.4%) Stapylococcus spp. isolates were identified as S. aureus, among which 200 (53.8%) possessed the mecA gene and were distinguished as MRSA. All of MRSA isolates contained blaZ gene. The frequency of ermA and ermC genes among erythromycin-resistant MRSA isolates was 21.6% and 66.7%, respectively. The frequency of the virulence genes eta, hla and sea among MRSA isolates was 10%, 80.5% and 100%, respectively. SCCmec type IV accounted for 30.6% of the MRSA isolates and SCCmec type III, SCCmec type II and SCCmec type I accounted for 30%, 22% and 17.5% of the isolates, respectively. The antibiotic resistance genes and the virulence genes of blaZ, hla, sea, eta and ermC had high frequencies among the MRSA isolates. This study showed that the antibiotic resistance genes had higher frequencies among SCCmec types I and IV, which confirms the previous reports in this field.

Emergence of SCCmec type III with variable antimicrobial resistance profiles and spa types among methicillin-resistant Staphylococcus aureus isolated from healthcare- and community-acquired infections in the west of Iran.

INTRODUCTION: Methicillin-resistant Staphylococcus aureus (MRSA) is a pathogen of public health importance. The prevalence of MRSA and its antimicrobial resistance pattern, as well as SCCmec and spa types, remain unclear both in the community and in the hospitals of the western region of Iran. METHODS: One hundred MRSA isolates were collected from different hospitals in the west of Iran during 2010-2011. Antimicrobial susceptibility testing to 15 antimicrobial agents was carried out by disk agar diffusion (DAD) method in accordance with the Clinical and Laboratory Standards Institute guidelines. Vancomycin minimum inhibitory concentrations (MICs) were evaluated by a broth microdilution method. The Etest was used for the detection of highly gentamicin-resistant MRSA. A combination of single and multiplex PCR was used for the detection of different resistance genes, including beta-lactamase, aminoglycoside modifying enzymes (AMEs), and macrolide-lincosamine, and for SCCmec typing of MRSA isolates. Genotyping of MRSA isolates was performed via spa typing. RESULTS: All tested isolates were susceptible to quinupristin-dalfopristin, linezolid, and vancomycin, but were resistant to penicillin (100%), erythromycin (50%), clindamycin (27%), and gentamicin (18%). MIC50 and MIC90 was 256 µg/ml among gentamicin-resistant MRSA. The most prevalent AME genes among aminoglycoside-resistant isolates were aac(6')-1e-aph(2")-1a (77.8%), aph(3')-IIIa (38.9%), and ant(4')-1a (27.8%). Nearly all tetracycline- and erythromycin-resistant MRSA had ermA and/or ermC but not ermB. Five SCCmec types and subtypes, 13 spa types, and four BURP groups (A-D) were identified. SCCmec types III (45%) and IVc (24%), spa type t701 (30%), and new spa type t12311 (15%) were the most prevalent among MRSA isolates. CONCLUSIONS: This study showed the emergence of MRSA with SCCmec type III and with spa types t12311, t10740, t1234, t1991, and t2651 with different phenotypic and genotypic antimicrobial resistance in the west of Iran. We found different SCCmec and spa types distributed among nosocomial and non-nosocomial MRSA in the west of Iran.

Phenotypic and genotypic characteristics of tetracycline resistant Acinetobacter baumannii isolates from nosocomial infections at Tehran hospitals.

OBJECTIVE(S): To date, the most important genes responsible for tetracycline resistance among Acinetobacter baumannii isolates have been identified as tet A and tet B. This study was carried out to determine the rate of resistance to tetracycline and related antibiotics, and mechanisms of resistance. MATERIALS AND METHODS: During the years 2010 and 2011, a total of 100 A. baumannii isolates were recovered from patients in different hospitals of Tehran, Iran. Antimicrobial susceptibility to tetracycline, minocycline, doxicycline and tigecycline was evaluated by E-test. Polymerase chain reaction (PCR) of the tet A and tet B genes was performed using specific primers, after which the isolates were subjected to Repetitive Extragenic Palindromic-PCR (PCR) to identify the major genotypes. RESULTS: Of all isolates, 89% were resistant to tetracycline (MIC50 = 32 µg/ml, MIC90 = 512 µg/ml). Minocycline with the resistant rate of 35% (MIC50 = 16 µg/ml, MIC90 =32 µg/ml) and doxicycline with the resistant rate of 25% (MIC50 = 16 µg/ml, MIC90= 32 µg/ml) have a good activity against A. baumannii isolates. All isolates were sensitive to tigecycline. Frequencies of tet B and tet A genes and coexistence of tet A and tet B among the isolates resistant to tetracycline, were 87.6%, 2.2% and 1.1%, respectively. Distribution of REP-types among A. baumannii isolates was types A (40%), B (30%), C (10%), D (5%) and E (5%). CONCLUSION: It seems that tet A and tet B genes play an important role in the induction of resistance towards tetracyclines used in this study. It is suggested that further studies focus on other antimicrobial drugs and combinations in order to achieve a successful therapy against multi drug resistance (MDR) A. baumannii strains in Iran.

Detection of highly ciprofloxacin resistance acinetobacter baumannii isolated from patients with burn wound infections in presence and absence of efflux pump inhibitor.

The emergence of fluoroquinolone resistance among A. baumannii isolates is now of particular concern. Phenotypic and genotypic characteristics of resistance to ciprofloxacin among 50 Acinetobacter baumannii isolated from burn wound infections of Tehran were evaluated by E-test and broth microdilution in presence and absence of efflux pump inhibitor phenylalanine- arginine ß-naphthylamide (PAßN) and PCR-sequencing methods. All isolates were then typed by REP-PCR fingerprinting to find the clonal relationship between resistant isolates. Our results indicated that resistance to ciprofloxacin among A. baumannii isolated from burn infections in Tehran are high with resistance rate of 100% and ciprofloxacin resistant isolates have a mutation of Serine 83 →Leucine in the quinolone resistance determining region (QRDR) of DNA gyrase subunit A (GyrA). 38% of the isolates showed MIC ranges of 64 to ≥512µg/ml and were considered as highly resistant. We could not detect Par C mutations and plasmid-mediated quinolone resistance A (qnrA) among ciprofloxacin resistant isolates. When we used the efflux pump inhibitor PAbN, MIC of ciprofloxacin was reduced two-to four folds. REP-type A (25/50; 50%), B (20/50; 30%) and C (10/50; 20%) were the most common REP-types among A. baumannii isolates. It seems that mutation in GyrA is the main mechanism of resistant to ciprofloxacin among A. baumannii isolates from burn infections and presence of efflux pumps is just secondary target for ciprofloxacin resistant among A. baumannii in Iran. Regarding with limitation of REP-types detected in this study, we found good correlation between resistance to ciprofloxacin and REP-types A-C.

Antimicrobial resistance patterns and their encoding genes among Acinetobacter baumannii strains isolated from burned patients.

The purpose of this study was to determine the mechanisms and patterns of antimicrobial resistance among the isolates obtained from burned patients with wound infections at a teaching hospital in Tehran, Iran. A total of 23 Acinetobacter baumannii isolates were collected from patients with burn wound infections between August 2009 and July 2010 from a hospital in Tehran. The susceptibility of these strains against 11 antimicrobial agents was determined by E-test according to the CLSI guidelines. All the resistant strains were then subjected to PCR assay for 28 distinct resistance genes. The most active antimicrobial agent was colistin with 100% sensitivity followed by gentamicin, amikacin and imipenem with 69.5%, 52.1% and 51.1% sensitivity, respectively. The most frequent resistance genes detected were bla(OXA-51-like) genes (n=23; 100%) that was intrinsic to A. baumannii isolates, gyrA (n=23; 100%), carO (n=23; 100%), tetA (n=22; 95.5%), tetB (n=15; 65.2%), intI (n=13; 56.5%) and PER (n=12; 52.1%), respectively. In order to make a proper choice of antibiotic for burn patients, it would be beneficial to physicians to identify drug resistance patterns in A. baumannii isolates.