Prevalence of high-level gentamicin-resistant Enterococcus faecalis and Enterococcus faecium in an Iranian hospital

This study was designed to determine the molecular characteristics and antimicrobial resistance of enterococcal strains isolated from patients admitted to an Iranian Hospital. Enterococcal strains were isolated from the burn patients. All strains were screened for genes encoding resistance to aminoglycoside aac(6')-Ie-aph(2'')-Ia, aph (3'), ant (4'), resistance to vancomycin (vanA, vanB), resistance to tetracycline (tetK, tetL, tetM, tetO), and resistance to erythromycin (ermA, ermB, ermC) by PCR and multiplex PCR-based methods. Genetic diversity was evaluated via Random Amplified Polymorphic DNA (RAPD)-PCR. All enterococcal isolates showed complete sensitivity to vancomycin with MIC � 0.5μg/ml. Resistance to gentamicin, tetracycline, erythromycin, ciprofloxacin or quinopristin-dalfopristin was detected, whilst more than 96.2% of isolates were high-level gentamicinresistant (HLGR) and multiple drug resistant. The most prevalent aminoglycoside resistance gene was aac(6')-Ie-aph(2'')-Ia, that was found in 96.2% (26/27) of the isolates. The most prevalent tetracycline resistance genes were tetM, found in 85.1% (23/27) followed by tetL and tetO found in 7.4% (2/27) of the isolates. The ermA and ermB genes were detected in 33.3% (9/27) and 44.4% (12/27) of the isolates respectively. RAPD-PCR analysis yielded 17 distinct profiles among 27 investigated isolates. One cluster of isolates shared the same RAPD pattern, while 16 isolates had unique RAPD pattern. Our study showed that during the examination time period one RAPD genotype was the common type and was disseminated among patients in the burn unit. Interestingly, most of these strains had an identical or very similar antibiotic and gene resistance pattern

High prevalence of direct repeat unit types of 10di, 8 h and 8i among methicillin resistant Staphylococcus aureus strains with staphylococcal cassette chromosome mec type IIIA isolated in Tehran, Iran 11 Medical and Health Sciences 1103 Clinical Sciences

Background: The emergence of methicillin-resistant Staphylococcus aureus (MRSA) is a main concern in burn care centers worldwide. The some reports of MRSA in Iran suggested that MRSA with type SCCmec III is common among burn patients. The aim of this study was to determine the prevalence, virulence genes, and antimicrobial susceptibility of the direct repeat units (dru) types of MRSA with SCCmec IIIA isolated from burn wounds in a burn care center in Tehran, Iran. Methods: In total, 165 S. aureus isolates were collected from clinical samples. In order to detect MRSA isolates, the mecA gene was amplified through the polymerase chain reaction (PCR) method. Antimicrobial susceptibility was tested using the disc agar diffusion test. Moreover, the PCR method was applied to determine SCCmec types, virulence genes, and antimicrobial resistance genes. The dru region was sequenced and thereby, dru types and dru repeats were identified. A similarity matrix was used to create minimum spanning tree (MST). Results: The prevalence of MRSA was 69 (114 out of 165 isolates). Most of MRSA isolates (61 out of 114, 53.5) were SCCmec type IIIA. All MRSA isolates were vancomycin-susceptible and more than 68 of MRSA isolates with SCCmec type IIIA were mupirocin resistant. The successful dru typing of isolates with SCCmec type IIIA revealed fourteen different dru types. There were two new dru types, namely dt10di and dt7aj. MST analysis indicated the presence of the three clusters of dt10di (cluster I), dt8i-dt8 h (cluster II), and dt11c-dt10ao-dt11dd-dt11a-dt10a (cluster III). There were significant differences between clusters I and II respecting antimicrobial resistance pattern and virulence genes. Conclusion: Three main dru clusters are prevalent in the study setting. The main dru types in the setting are dt10di, dt8i, and dt8 h. Dru typing can be used to differentiate MRSA strains with SCCmec IIIA. © 2019 The Author(s)

Efficacy of 16S rRNA variable regions high-resolution melt analysis for bacterial pathogens identification in periprosthetic joint infections

Background: Accurate and rapid identification of microorganisms causing periprosthetic joint infections (PJIs) are necessary for choosing an appropriate antibiotic therapy. Therefore, molecular techniques are suggested for diagnosis in suspected PJIs. The Broad-range PCR and High-Resolution Melt Analysis (HRMA) were evaluated for the identification of causative organisms of PJIs in this study. Results: For 47 of 63 specimens, both the culture and broad-range PCR were positive. The culture was found to be able of organism�s detection in 74.6 (47/63) of patients. Of 47 positive cultures, 11 (23.4) were polymicrobial and 36 (76.59) were monomicrobial cultures, in which 34 (91.89) cases were detected by HRM assay. The sensitivity, specificity of HRMA vs monomicrobial culture were 91.89, 93.75, respectively. The sensitivity, specificity of total HRMA (mono + poly) vs culture were 82.92, 93.75. Conclusions: HRM assay coupled with broad-range PCR are effective screening, rapid, and relatively cost-effective methods for discrimination of PJIs especially in aiding culture method. Using computer programs such as the Matlab-2018b program for HRM data analysis is also valuable and helpful in diagnosis. © 2021, The Author(s)

VIRULENCE FACTORS, ANTIMICROBIAL SUSCEPTIBILITY AND MOLECULAR CHARACTERIZATION OF STREPTOCOCCUS AGALACTIAE ISOLATED FROM PREGNANT WOMEN

Forty-one Streptococcus agalactiae isolates collected from pregnant women at 35-37 weeks of gestation were analysed for their capsular types, antimicrobial resistance determinants, distribution of virulence factors and genetic relatedness using PCR and multiplex PCR. Capsular type III was predominant (65.8), followed by capsular type II (14.6), Ib (7.3), and V(4.9). All isolates were susceptible to penicillin, vancomycin, linezolid and quinupristin-dalfopristin. Resistance to tetracycline, erythromycin and clindamycin were found in 97.6, 24.4, and 14.6 of isolates, respectively. The most common antimicrobial resistance gene was tetM found in 97.6 of the isolates followed by ermTR and ermB found in 12 and 7.3 of isolates, respectively. The most common virulence gene was hly (100), followed by scpB (97.6), bca (97.6), rib (53.65) and bac (4.9). The insertion sequence IS1548 was found in 63.4 of isolates. By multi locus variable number of tandem repeat analysis (MLVA) typing, 30 different allelic profiles or MLVA types (MTs) were identified. The most frequent was the MT1 (5/41, 12.2) and followed by MT2 (4/41, 9.75). Our data revealed that population structure of these isolates is highly diverse and indicates different MLVA types

Investigation of biofilm formation ability, antimicrobial resistance and the staphylococcal cassette chromosome mec patterns of methicillin resistant Staphylococcus epidermidis with different sequence types isolated from children

This study investigated the molecular characterizations of 80 methicillin resistant Staphylococcus epidermidis (MRSE) collected during 2012-2013 in Tehran Children's Medical Center, Iran. About 90 of MRSE isolates were multi-drug resistant (MDR) and the highest resistance was observed to cotrimoxazole and they were quite sensitive to quinupristin-dalfopristin and linezolid. Though vanA gene was not detected, the majority of isolates showed intermediate resistance to vancomycin (MIC90 16 mu g/ml). Resistance to mupirocin was observed in 18 isolates. Staphylococcal cassette chromosome mec (SCCmec) types V, III, IV and II were detected in 23.75, 7.5, 6.25 and 5 of isolates respectively, in some of which the additional parts of mec or ccr complexes were observed. In 57.5 MRSE isolates SCCmec types were not classified. 41.2 of MRSE isolates were carrying intercellular adhesion (ica) operon and 40 had strong or intermediate biofilm. The types of arginine catabolic mobile element (ACME) were limited to type I and II. Nine sequence types (STs) were seen in mupirocin resistant MRSE isolates. The common STs were ST2, ST5 and ST22 with 27.7 (5/18), 22.2 (4/18) and 16.6 (3/18) frequencies, respectively. ST23, ST54 and ST179 plus three novels STs 580, 581,588 were also observed. The majority of STs, 83.3 (15/18) belonged to clonal complex 2 (CC2). The spread of antibiotic resistance and virulence factors among MRSE species is an alarming sign in Children's Hospitals. The combination of these two issues leads to increase the chance of successfully establishing of common STs in hospital environments, and promotes the device-related infections and bacteremia. (C) 2016 Elsevier Ltd. All rights reserved

Characterization of virulence factors, antimicrobial resistance pattern and clonal complexes of group B streptococci isolated from neonates

Between January and December 2013, swab samples were taken for the throat and external ear canals of 1037 newborns for screening of Group B Streptococcus (GBS or S. agalactiae). Isolates were analyzed form Multilocus sequence typing (MLST), capsular type, virulence genes and antibiotic susceptibility. The MLST analysis of 19 GBS isolates showed 8 sequence types (STs). Overall the most common STs were ST19 and ST28. Other STs were ST1, ST4, ST8, ST12, ST335 and ST734 (a new ST). The most common clonal complexes (CCs) were CC19 (68.4) and CC10 (21). The scpB, hlyB and bca virulence genes were detected in all STS, while the bac gene was predominant in ST12 with capsular type (CT) Ib. The IS1548 and the rib genes were particularly prevalent in CTIII and were detected in isolates belong to ST19, ST335 and ST734 and were grouped in CC19. All isolates were susceptible to penicillin, vancomycin, linezolid and quinupristin-dalfopristin. Resistance to tetracycline was observed in all 19 (100) strains and was correlated with presence of the tetM gene except for one isolate with ST12. All the ST8 and ST12 isolates were resistant to macrolide carrying two resistance genes; the ermTR and the ermB, respectively. The results of this study showed that the CC19 was a major clone in the neonatal intensive care unit (NICU) of Imam Khomeini hospital which can cause severe infections in susceptible neonates (particularly in premature infants). As a result, an intensive infection control policy is needed to prevent the spread of this clone. © 2016 Elsevier Lt