Detection Of mecA Gene Research Articles

A simple and sensitive colorimetric approach for mecA gene analysis via exonuclease-III catalyzed signal cascade

Analysis of mecA gene in Staphylococcus aureus (S. aureus) is essential for controlling infections in intensive care units (ICU) and preventing the use of ineffectual empirical treatments. However, quantitative determination of the mecA gene remains difficult. Herein, we propose a simple and sensitive colorimetric approach by integrating exonuclease-III (Exo-III) assisted signal cascade and G-quadruplex/hemin DNAzymes (G4 DNAzymes) catalyzed 2,2′-azino-bis (3-ethylben-zothiazoline-6-sulfonic acid) (ABTS) based color reaction. In this method, signal amplification does not necessitate the use of complex experimental components, such as multiple enzymes and primer design, while still maintaining a high signal amplifying efficiency. Therefore, the method has a broad mecA gene detection range from 10 fM to 1 nM and a low limit of detection down to 3.4 fM level. Taking the merit of simplicity and high sensitivity, the approach is promising in analyzing mecA gene in S. aureus and diagnosing infections.

Molecular Detection of mec-A Gene and Antimicrobial Profiling of Clinically Associated Staphylococcus aureus

Molecular Detection of mec-A Gene and Antimicrobial Profiling of Clinically Associated Staphylococcus aureus

Molecular prevalence of MecA and Blaz Genes with phenotypic analysis of Antibiotic Sensitivity Pattern for S.aureus Isolated From Dermal lesions of Sheep Breeders In Diyala Governorate – Iraq

Background: S. aureus is one of the dominant bacterial pathogens among dermal infections in human and animals, which have resistance to different antimicrobial drugs.
 Objective: Isolation and identification of S.aureus from skin lesions among sheep breeders by traditional methods, Vitek 2 system and PCR using Staur 4, 6 specific pri-mers for validation, Detection of genes (methicillin resistant (mecA), Beta- lac-tamase gene (blaZ) by conventional PCR ,determine antibiotic sensitivity pat-tern by kirby bauer disc diffusion method.
 Patients and Methods: A total of 44 swaps were collected from sheep breeders suffered from variety of infected skin lesions to detect methicillin sensitive and resistant s. aureus by employing traditional methods in addition to confirmatory techniques through fast rapid VETEK2 system , detection of genes (methicillin resistant (mecA), Be-ta- lactamase gene (blaZ) by conventional PCR , and determine antibiotic sensi-tivity pattern by kirby bauer disc diffusion method.
 Results: S.aureus was isolated from 15/44,( 34.09%) of skin lesion of sheep breeders . A total of 6/15 ,(40%) were methicillin resistant S.aureus (MRSA) ,which represent ( 13.63%) from total samples .Beta lactamase gene primers was detected in all S.aureus isolates. A 6/15,(40%) of S.aureus have resistance for members of antibiotics classes, penicillins, polypeptides, fluoroquinolones, macrolide which include the following : methicillin that was confirmed early by detection of me-cA gene , levofloxacin , ofloxacin , erythromycin and vancomycin. A 6/15,(40%) of S.aureus have multidrug resistant trait for Penicillins, Polypep-tides, Fluoroquinolones, Macrolide antibiotics. Non multidrug resistant S.aureus was reported for Penicillins (oxacillin,4/15, 26%) and Polypeptides antibiotics (vancomycin 8/15, 53.33%) of S.aureus . Absolute sensitivity was reported for gentamycin, tetracycline, rifampicin, imipenem and chloramphenicol.
 Conclusion: Methicillin sensitive S.aureus was more common compared with MRSA isolated from dermal infections of sheep breeders. Blaz gene was predominantly ex-pressed by S.aureus isolates followed by Mec A gene. S.aureus have resistance for penicillins, polypeptides, fluoroquinolones, macrolide which include the fol-lowing : methicillin, levofloxacin , ofloxacin , erythromycin and vancomycin . S.aureus have multidrug resistant trait for Penicillins, Polypeptides, Fluoroquin-olones, Macrolide antibiotics. S.aureus was absolutely sensitive to gentamycin, tetracycline, rifampicin, imipenem and chloramphenicol.

Molecular prevalence of MecA and Blaz Genes with phenotypic analysis of Antibiotic Sensitivity Pattern for S.aureus Isolated From Dermal lesions of Sheep Breeders In Diyala Governorate – Iraq

Background: S. aureus is one of the dominant bacterial pathogens among dermal infections in human and animals, which have resistance to different antimicrobial drugs. Objective: Isolation and identification of S.aureus from skin lesions among sheep breeders by traditional methods, Vitek 2 system and PCR using Staur 4, 6 specific pri-mers for validation, Detection of genes (methicillin resistant (mecA), Beta- lac-tamase gene (blaZ) by conventional PCR ,determine antibiotic sensitivity pat-tern by kirby bauer disc diffusion method. Patients and Methods: A total of 44 swaps were collected from sheep breeders suffered from variety of infected skin lesions to detect methicillin sensitive and resistant s. aureus by employing traditional methods in addition to confirmatory techniques through fast rapid VETEK2 system , detection of genes (methicillin resistant (mecA), Be-ta- lactamase gene (blaZ) by conventional PCR , and determine antibiotic sensi-tivity pattern by kirby bauer disc diffusion method. Results: S.aureus was isolated from 15/44,( 34.09%) of skin lesion of sheep breeders . A total of 6/15 ,(40%) were methicillin resistant S.aureus (MRSA) ,which represent ( 13.63%) from total samples .Beta lactamase gene primers was detected in all S.aureus isolates. A 6/15,(40%) of S.aureus have resistance for members of antibiotics classes, penicillins, polypeptides, fluoroquinolones, macrolide which include the following : methicillin that was confirmed early by detection of me-cA gene , levofloxacin , ofloxacin , erythromycin and vancomycin. A 6/15,(40%) of S.aureus have multidrug resistant trait for Penicillins, Polypep-tides, Fluoroquinolones, Macrolide antibiotics. Non multidrug resistant S.aureus was reported for Penicillins (oxacillin,4/15, 26%) and Polypeptides antibiotics (vancomycin 8/15, 53.33%) of S.aureus . Absolute sensitivity was reported for gentamycin, tetracycline, rifampicin, imipenem and chloramphenicol. Conclusion: Methicillin sensitive S.aureus was more common compared with MRSA isolated from dermal infections of sheep breeders. Blaz gene was predominantly ex-pressed by S.aureus isolates followed by Mec A gene. S.aureus have resistance for penicillins, polypeptides, fluoroquinolones, macrolide which include the fol-lowing : methicillin, levofloxacin , ofloxacin , erythromycin and vancomycin . S.aureus have multidrug resistant trait for Penicillins, Polypeptides, Fluoroquin-olones, Macrolide antibiotics. S.aureus was absolutely sensitive to gentamycin, tetracycline, rifampicin, imipenem and chloramphenicol.

Detection of positive mecA Staphylococcus aureus isolated from meat and butchers’ shops by using PCR technique in Mosul city

Detection of positive mecA Staphylococcus aureus isolated from meat and butchers’ shops by using PCR technique in Mosul city

Antimicrobial resistance and its detection in Staphylococcus saprophyticus urinary isolates

The aim of this study was to describe the antibiotic susceptibility of clinical Staphylococcus saprophyticus isolates collected prospectively from urine specimens over a 2-month period from September to October 2022 at a single centre in Melbourne, Australia. Species identification was performed by MALDI-TOF MS. All isolates underwent phenotypic antibiotic susceptibility testing by disc diffusion using European Committee on Antimicrobial Susceptibility Testing (EUCAST) and Clinical and Laboratory Standards Institute (CLSI) guidelines and VITEK2, and mecA polymerase chain reaction. A total of 302 S.saprophyticus isolates from 298 patients were included in this study. Most specimens (91.1%) were referred by community general practitioners from non-hospitalised patients. Antimicrobial resistance to non-β-lactam antibiotics was uncommon; trimethoprim susceptibility was 97%; trimethoprim/sulfamethoxazole, 98%; nitrofurantoin, 100%; and ciprofloxacin, 100% (100% ciprofloxacin susceptible, increased exposure by EUCAST breakpoints). Methicillin resistance (by mecA detection) was the most common form of urinary antibiotic resistance at 5.6%. VITEK2 susceptibility testing for methicillin resistance had a poor specificity of 61.8% (95% CI 55.8-67.4%) compared to mecA detection. These findings indicate that empiric antibiotic recommendations of trimethoprim, trimethoprim/sulfamethoxazole, and nitrofurantoin for treatment of urinary S.saprophyticus remain appropriate.

Dual-Toehold-Probe-Mediated Exonuclease-III-Assisted Signal Recycles Integrated with CHA for Detection of mecA Gene Using a Personal Glucose Meter in Skin and Soft Tissue Infection.

Staphylococcus aureus integrated with mecA gene, which codes for penicillin-binding protein 2a, is resistant to all penicillins and other beta-lactam antibiotics, resulting in poor treatment expectations in skin and soft tissue infections. The development of a simple, sensitive and portable biosensor for mecA gene analysis in S. aureus is urgently needed. Herein, we propose a dual-toehold-probe (sensing probe)-mediated exonuclease-III (Exo-III)-assisted signal recycling for portable detection of the mecA gene in S. aureus. When the target mecA gene is present, it hybridizes with the sensing probe, initiating Exo III-assisted dual signal recycles, which in turn release numerous "3" sequences. The released "3" sequences initiate catalytic hairpin amplification, resulting in the fixation of a sucrase-labeled H2 probe on the surface of magnetic beads (MBs). After magnet-based enrichment of an MB-H1-H2-sucrase complex and removal of a liquid supernatant containing free sucrase, the complex is then used to catalyze sucrose to glucose, which can be quantitatively detected by a personal glucose meter. With a limit of detection of 4.36 fM for mecA gene, the developed strategy exhibits high sensitivity. In addition, good selectivity and anti-interference capability were also attained with this method, making it promising for antibiotic tolerance analysis at the point-of-care.

Diversity of <em>Spa</em> gene between methicillin-resistant and methicillin-sensitive <em>Staphylococcus aureus</em> bacteria in a tertiary referral hospital, Indonesia

BACKGROUND Spa typing is an effective and fast technique to identify the prevalence and spread of Staphylococcus aureus strains based on their spa gene profiles. The distribution of spa types will contribute to control the spread of S. aureus. Little is known regarding the spa types of S. aureus in Indonesia. This study aimed to investigate the diversity of spa gene among S. aureus carriage isolates in North Sumatra Province, Indonesia.
 METHODS 79 S. aureus isolates consisting of 39 methicillin-resistant S. aureus (MRSA) and 40 methicillin-susceptible S. aureus (MSSA) carriage isolates were identified by VITEK2 Compact (Biomerieux, Indonesia) to detect mecA gene. All samples underwent spa typing and sequencing.
 RESULTS Spa gene was detected among 31/39 (79%) of the MRSA isolates and 24/40 (60%) of the MSSA isolates. Most spa typing genes were identified between 350 and 400 base pair (bp). t258 and t852 were the most prevalence spa types among MRSA and MSSA isolates, respectively.
 CONCLUSIONS Many MRSA and MSSA isolates encoded spa gene. The most genes detected were t258 and t852, identified in Germany and Portugal, respectively; while t18977 was initially identified in Malaysia. This indicated a global spread of MRSA according to spa typing.

Antibiogram of Staphylococcal Isolates and Molecular Characterisation of Coagulase Negative Staphylococci Obtained from a Hospital in Port Harcourt, Nigeria

Difficulty in the treatment and eradication of bacterial infections as a result of the development of resistance to antibiotics has in recent time remained a serious problem confronting public health. This study investigated the antibiogram of Staphylococcal isolates and molecular characterisation of Coagulase Negative Staphylococci (CoNS) obtained from female patients visiting Rivers State University Teaching Hospital (RSUTH) in Port Harcourt, Nigeria. A total of 103 clinical samples were collected and processed using the standard laboratory procedures. Antibiotic susceptibility test of the isolates was performed using Kirby-Bauer disc diffusion method, following Clinical and Laboratory Standards Institute (CLSI) guidelines. Molecular characterisation as well as screening for the detection of antibiotics resistance MecA gene was carried out using Polymerase Chain Reaction (PCR) dependent molecular techniques. From the study, 22.3% S. aureus and 1.9% CoNS were recovered. The 16SrRNA gene sequencing revealed two species of CoNS namely S. saprophyticus and S. arlettae. The antibiotics susceptibility test results revealed that all the Staphylococcus aureus isolates were 100% resistant to six of the antibiotics tested: Ampicillin, Augmentin, Ceftazidime, Cefuroxime and Cephalexin (β- lactams), 78.3% resistant to Tetracycline and Cotrimoxazole, 73.9% to Vancomycin, 65.2% to Ciprofloxacin, 56.5% to Gentamicin and 47.8% to Erythromycin but 52.2% sensitive to Erythromycin, followed by Gentamicin (34.8%), Ciprofloxacin (17.4%), Vancomycin and Cotrimoxazole (13%) and Tetracycline (8.7%). All (100%) of the Coagulase negative Staphylococci species were resistant to every antibiotics tested. Data obtained from the study showed all the isolates displayed multidrug resistance, with MAR indices greater than 0.2 and Methicillin (MecA) gene was detected in 66.6% of the isolates. The study has reported the presence of Coagulase Negative Staphylococci showing resistance to all the tested antibiotics, hence indicating alarming public health threats. Strict regulation of antimicrobial usage is therefore vital.

Role of Oxacillin Susceptibility Testing Strategy in Changing Scenario of mecA Positive Staphylococcus aureus Isolates (OS-MRSA) Detection

Staphylococcus aureus strains that are mecA and PBP2a positive but phenotypically susceptible to oxacillin are becoming more and more abundant, according to research from all around the world. The oxacillin susceptibility of Staphylococcus aureus (OS-MRSA) contributes to consequent treatment-failure due to misidentification by conventional susceptibility tests. Therefore, the objective of the current study was to ascertain the prevalence of OSMRSA in a tertiary care facility located in Mysore, South India. 395 MRSA isolates collected from diverse clinical samples were included in this lab-based prospective investigation. These isolates were tested using an oxacillin 1μg disc phenotypically by standard disc diffusion test, and simultaneously MIC to Oxacillin was determined from Vitek2 systems. Additionally, MRSA specific mecA gene detection was applied to these isolates in order to confirm their MRSA status genotypically. PCR findings demonstrate that 65% of the isolates were MRSA. The vitek2 system detected 4.06% OS-MRSA isolates with an oxacillin MIC of ≤2µg/ml. The disc diffusion method identified a total of 13.75% isolates as oxacillin sensitive and 10% isolates were oxacillin intermediately sensitive. Oxacillin sensitivity was shown for 1.87% of the mecA-positive MRSA isolates using the VITEK2 and disc diffusion techniques. This analysis found isolates with lower oxacillin MICs but relatively reduced OS-MRSA incidence. Using an oxacillin disc for routine laboratory MRSA detection might occasionally produce false negative results, which can result in improper antibiotic administration and treatment failure. In order to distinguish OS-MRSA from MRSA, it is crucial to combine phenotypic and genotypic techniques.

Emergence of clinical <i>vanA</i>-type vancomycin-resistant <i>Staphylococcus aureus</i> isolates in National Orthopaedic Hospital Dala, Kano, Nigeria

Background: The increasing prevalence of multi drug resistance (MDR) in strains of Staphylococcus aureus is a major challenge in the selection of an appropriate therapeutic agents, especially in persistent orthopaedic infections. This study investigated the patterns of antimicrobial resistance and identified the genetic determinants of resistance in S. aureus isolates from orthopaedic patients.
 Methodology: This was a descriptive cross-sectional study of hospitalized patients at National Orthopaedic Hospital Dala (NOHD), Kano, Nigeria from whom urine samples, and nasal and wound swabs were collected for isolation of S. aureus. Samples were cultured on standard media and S. aureus isolated and identified using both conventional biochemical tests and a standard rapid diagnostic kit. The antibiotic susceptibility was determined to a panel of 15 antibiotics using the modified Kirby-Bauer disc diffusion method. Vancomycin minimum inhibitory concentration (MIC) of each isolate was determined using vancomycin Epsilon-test strip. mecA and vanA were detected by multiplex polymerase chain reaction (PCR) assay.
 Results: From the total of 134 samples, S. aureus was isolated from 36 (26.8%); 10 (7.4%) from urine, 13 (9.7%) from nasal swab, and 13 (9.7%) from wound swab. Thirty-four (94.4%) isolates were phenotypically methicillin (cefoxitin) resistant (MRSA), while 2 (5.6%) isolates were methicillin sensitive (MSSA). Phenotypic resistance rate of the S. aureus isolates was highest to gentamicin (94.4%), followed by penicillin (88.8%), cephalosporins and fluoroquinolones (87.4%), while rate was lowest to vancomycin (11.1%, 4/36). Seventeen (47.2%) were MDR, 16 (44.4%) were extensively drug resistant (XDR), and 2 (5.6%) were pan-drug resistant (PDR) S. aureus isolates. The mecA gene was detected in 4 (11.8%) of the 34 phenotypic MRSA isolates and vanA genes in 2 (50.0%) of the 4 phenotypic VRSA isolates.
 Conclusion: The detection of vanA and mecA in clinical S. aureus isolates in this study is an indication that clinical VRSA has emerged in MRSA population in Nigeria. This emergence can pose a major threat to primary care-givers and a public health challenge among the daily inhabitants of National Orthopaedic Hospital Dala (NOHD), Kano and the community at large.

Comparison of cefoxitin disc diffusion test, HiCrome rapid MRSA agar and PCR for<em> mec</em>A, PVL and <em>fem</em>B genes for detection of methicillinresistant <em>Staphylococcus aureus</em> (MRSA)

This study evaluates the effectiveness of cefoxitin disk diffusion as a method to detect methicillin-resistant Staphylococcus aureus (MRSA), comparing with HiCrome MRSA screening agar and PCR-based mecA gene detection. Both PCR and the cefoxitin disk diffusion test exhibited 100% sensitivity and specificity, while HiCrome agar showed slightly lower sensitivity (90%) and specificity (95.24%) out of 72 Staphylococcus aureus isolates. The MRSA detection rate was found to be 41.66%, with femB and PVL genes present in 86.6% and 13.3% of MRSA isolates, respectively. Cefoxitin could serve as a reliable substitute marker for MRSA detection, particularly in settings with limited resources.

Prevalence and clonal diversity of methicillin-resistant Staphylococcus aureus andmethicillin-resistant Staphylococcus pseudintermedius isolated from dogs andcatswith eye discharge.

Methicillin-resistant Staphylococcus (MRS) has originated, spread extensively, and become a prominent source of bacterial infections in both human and animal. We report the prevalence, genetic diversity, and antimicrobial resistance pattern of Staphylococcus pseudintermedius and Staphylococcus aureus strains isolated from dogs and cats with eye discharges. A total of 12 (6.0%) coagulase-positives staphylococci were identified as (6/200, 3%) S. aureus and (6/200, 3%) S. pseudintermedius. The phenotypic methicillin resistance of S. aureus and S. pseudintermedius were 50.0% (3/6) and 16.7% (1/6), respectively. None of the isolates showed biofilm formation in the microtiter plate assay. The highest resistance (50.0%) for S. pseudintermedius strains was detected against clindamycin and tetracycline. 67.0% of S. aureus isolates were resistant to penicillin-G. The PCR analysis conducted for detection of mecA gene indicated that only one S. aureus isolated from a cat was mecA gene positive. Phylogenetic analysis based on repetitive sequence-based PCR (rep-PCR) showed that all strains were typable and generated PCR products ranging from 800 bp to 4,400 bp. The lineages ST241 and the novel ST2361 in multi-locus sequence typing (MLST) analysis were detected in one methicillin-susceptible S.pseudintermedius and methicillin-resistant S. pseudintermedius of dogs, respectively. Inaddition, the lineages ST4155 and ST7217 of two methicillin-resistant S. aureus strains of cats were connected epidemiologically to previously reported cases. These results indicate epidemiologically related strains (ST241, ST4155, and ST7217) transferring between animals and humans. Therefore, the strategies to combat the widespread MRS should be based on collaboration between human and veterinarymedicine under the One Health concept.

Molecular characterization and epidemiology of methicillin-resistant Staphylococcus aureus isolated from clinical samples in Sokoto, Nigeria

Objectives: Methicillin-resistant Staphylococcus aureus (MRSA) is a major public health threat and a cause of hospital-acquired and community-acquired infections. This study was undertaken to investigate antimicrobial resistance pattern, the genetic lineage, presence of S. aureus protein A (SPA) gene, and staphylococcal chromosomal cassette mec (SCC mec) types of MRSA isolated from clinical samples sent for microbiological test in major government hospitals in Sokoto. Material and Methods: A total of 90 S. aureus MRSA isolates were collected and confirmed using standard microbiological techniques. Antibiotic susceptibility testing was done using the disk diffusion method; mecA detection and sequencing were carried out. Phylogenetic reconstruction was also done using the molecular evolutionary genetics analysis X software and phylogeny tree constructed by Neighbor-Joining method. SCC mec typing and SPA detection were also done. Results: Of the 90 S. aureus isolates, 42 were found to be MRSA using the cefoxitin disk diffusion, the most potent antibiotic against them was quinupristin/dalfopristin with 83.3% followed by rifampicin with 81.0% and 6 clindamycin with a 71.4%. With 78.6% of the isolates showing resistance to the fluoroquinolone antibiotic ciprofloxacin, tetracycline and gentamicin came in second and third, with 64.3% and 61.9% of isolates showing resistance, respectively. Most of the MRSA isolates were resistant to more than three antibiotics. Polymerase chain reaction showed 36 (85.7%) harbored the mecA gene and of the 36 mecA positive isolates, phylogenetic reconstruction of representative MRSA sequences showed that MRSA sequences in this study clustered in two closest clades suggesting a possible horizontal transfer. Of the 36 isolates, 23 were SCC mec type I, ten were type IV, and three were non-typeable, while the SPA gene was detected in all the isolates amplified. Conclusion: The use of phenotypic and molecular methods in this study provided useful information on antibiotic resistance profile, epidemiology, and molecular characteristics of MRSA isolates in Sokoto Nigeria. The information provided could help in monitoring the evolution of MRSA strains in Nigeria over time.

ANTIBIOTIC RESISTANCE AND MOLECULAR DETECTION OF nuc AND mecA GENES OF STAPHYLOCOCCUS AUREUS ISOLATED FROM MASTITIC MILK

The emergence of methicillin resistant Staphylococcus aureus (MRSA) is mainly due to the presence of resistant genes. The objective of this study is to perform antibiotic susceptibility profiling and molecular detection of nuc and mecA genes of Staphylococcus aureus isolated from mastitic milk. A total of 105 milk samples were collected from dairy farms in the Faisalabad region. They were cultured on Staph-110 media and mannitol salt agar. Biochemical tests included catalase and coagulase, were also performed for the identification of Staphylococcus aureus. Antibiotic susceptibility profiling of MRSA was done by disc diffusion method and molecular screening of Staphylococcus aureus for the detection of nuc and mecA genes was done by using PCR. Selected 8 antibiotics based on previous history being commonly used in dairy practices. The prevalence of Staphylococcus aureus observed by PCR was 58.09%. Oxacillin (1µg) and vancomycin (30µg) showed 100% resistance, while enrofloxacin (5µg), amoxicillin (25µg), ampicillin (10µg), oxytetracycline (1µg), gentamycin (10µg) and tylosin (30µg) showed 1.64, 55.74, 73.77, 13.11, 1.64 and 11.74% resistance, respectively. Susceptibility percentages of enrofloxacin (5µg), amoxicillin (25µg), ampicillin (10µg), oxytetracycline (1µg), gentamycin (10µg) and tylosin (30µg) were 27.87, 37.70, 14.75, 19.67, 98.36 and 9.84%, respectively. Enrofloxacin (5µg), amoxicillin (25µg), ampicillin (10µg), oxytetracycline (1µg) and tylosin (30µg) showed 70.49, 6.56, 11.47, 67.21 and 78.69% intermediate resistance against MRSA. Among the mastitic cases, 58.09% were positive for Staphylococcus aureus. These isolates were susceptible to gentamycin in 98.36% and tylosin in 78.69% cases, so these antibiotics can be used for the treatment of mastitis.

Detection of mec A gene in Staphylococcus aureus isolated from wounds infections

This study aimed to detect the occurrence of mecA gene in S.aureus isolates by PCR technique. The study included a total of 25 S. aureus was collected from patients with a wound infection. Identification of S. aureus was based on growth on manitol salt agar, catalase test, DNase test and coagulase test. Antibiotic Susceptibility test by disc diffusion method was done for all S. aureus isolates against four β- lactam antibiotics and the resistance percentage was as the following: 92% for methicillin, 32% for oxacillin, 60% for amoxicillin –clavulanic acid and 24% for ampicillin – sulbactam. PCR technique was used for the detection of mecA gene in S. aureus isolates and results showed that 24% of isolates were mecA positive, while 76% of the isolates were mecA negative

Prevalence of mecA and Panton-Valentine Leukocidin Genes in Staphylococcus aureus Clinical Isolates from Gaza Strip Hospitals.

Methicillin-resistant Staphylococcus aureus (MRSA) are spreading worldwide in hospital and community settings, thus posing a serious public health problem. Panton-Valentine Leukocidin (PVL), an important virulence factor of S. aureus, is a marker of community-acquired MRSA. Here we determined the prevalence of pvl genes among S. aureus isolates from different hospitals in the Gaza Strip, Palestine. A total of 285 S. aureus isolates were collected from five different hospitals in the Gaza Strip. All isolates were characterized for their susceptibility patterns to available antimicrobial agents and by using multiplex PCR for the detection of mecA and pvl genes. The overall prevalence of MRSA in Gaza hospitals was 70.2% (range: 76.3% to 65.5%) and that of pvl among S. aureus isolates was 29.8% (range: 32.9% to 26.2%). The pvl gene was equally prevalent among MRSA isolates (30.5%) and MSSA isolates (28.2%). The most effective antibiotics were rifampicin, vancomycin, and clindamycin, with susceptibility rates of 91.2%, 88.7%, and 84.6%, respectively. The highest percentage of strains were observed to be resistant to penicillin and amoxicillin with clavulanic acid-96.1% and 73.6%, respectively. Our results showed a high prevalence of MRSA and pvl-positive isolates in Gaza Strip hospitals, which likely reflects the situation in the community. It is mandatory to implement systematic surveillance of both hospital and community isolates, together with interventions (such as increased hand hygiene, use of hydroalcoholic solutions, and isolation of carriers) to limit their spread.

Antibiotic Resistance and Detection of Blatem and MecA Genes in Bacteria Isolated from Street Vended Pounded Yam in Yenagoa, Nigeria

Antibiotic resistant pathogens spread through food are a public health concern. The aim of this study was to determine the profile of antibiotic resistance and to investigate the presence of genes that produce antibiotic resistance in bacterial isolates from pounded yam collected from five sites in Yenagoa, Nigeria (Swali, Amarata, Kpansia, Tombia, and Akenfa). The Kirby-Bauer disk diffusion method was used to examine antibiotic susceptibility to nine antibiotics (Augmentin, Ofloxacin, Chloramphenicol, Ampicillin, Erythromycin, Gentamycin, Nitrofuratoin, Streptomycin and Tetracyclin), and the PCR method was utilized to find the blaTEM and mecA genes. A total of 150 pounded yam samples were collected and analyzed. Shigella spp, Bacillus spp, Staphylococcus spp, Pseudomonas spp, Salmonella spp, Klebsiella spp, Proteus spp, and Escherichia coli are among the bacteria that were recovered from the pounded yam samples. Except for Pseudomonas spp., all tested positive for multidrug resistance (resistance to three or more tested antibiotics), with the majority of these antibiotics being Chloramphenicol, Ampicillin, Erythromycin, Gentamycin, Nitrofuratoin, Streptomycin, and Tetracyclin. The Escherichia coli isolated from the street-vended pounded yam exhibited multidrug resistance against Ampicillin (37.5%), Erythromycin (43.8), Gentamycin (34.4), Streptomycin (21.9) and Tetracyclin (40.6). Bacillus spp also showed multidrug resistance against Chloramphenicol (36.4), Ammpicillin (36.4), Erythromycin (45.5), Gentamycin (36.4), Nitrofuratoin (36.4), Streptomycin (27.3) and Tetracyclin (36.4). Inhibition zones against Augmentin, Ofloxacin, Chloramphenicol, Ampicillin, Erythromycin, Gentamycin, Nitrofuratoin, Streptomycin, and Tetracyclin were clearly visible in Pseudomonas spp (100, 96.4, 64.3, 50.0, 82.1, 75.0, 64.3, 92.9, and 50.0% respectively). The highest resistance by number of bacterial isolates was found in Erythromycin (6 isolates showed resistance), followed by Tetracyclin (5 isolates showed resistance). Ampicillin and Streptomycin resistance was present in four isolates. Three representative isolates were selected for molecular identification of blaTEM and mecA which were amplified at 600bp and 500bp respectively. These genes were responsible for the antibiotic resistance seen in the isolates. According to this study, the pounded yam samples that were evaluated had food-borne disease strains that are multidrug resistant and a danger to the general public's health. The findings cast doubt on the quality of foods sold on the streets of Yenagoa, Nigeria.

Detection of mecA and PVL-Resistant Genes in Staphylococcus aureus from Clinical Samples in Bayelsa State, Nigeria

Detection of mecA and PVL-Resistant Genes in Staphylococcus aureus from Clinical Samples in Bayelsa State, Nigeria

Risk Factors, Genetic Diversity, and Antimicrobial Resistance of Staphylococcus spp. Isolates in Dogs Admitted to an Intensive Care Unit of a Veterinary Hospital.

Intensive Care Units (ICU) usually provide an excellent environment for the selection of pathogens associated with hospital-acquired infections (HAI), leading to increased mortality and hospitalization costs. Methicillin-resistant Staphylococcus pseudintermedius (MRSP) is a major cause of HAI in dogs worldwide, but the risk factors and dynamics of colonization by MRSP are largely unknown. This study aimed to evaluate the risk factors associated with the acquisition of MRSP in dogs admitted to an ICU, and to report the antimicrobial resistance profiles and genetic relatedness of MRSP isolates. Sterile swabs from the nostril, axilla, and rectum were collected daily during the hospitalization of 54 dogs. Samples were subjected to Mannitol Salt Agar, and colonies were identified by MALDI-ToF, polymerase chain reaction (PCR), and sequencing of the rpoB gene. Antimicrobial susceptibility testing and PCR detection of mecA were performed. Staphylococcus spp. was isolated from 94% of the dogs, and the most frequently isolated species was S. pseudintermedius (88.2%). Carriage of multidrug resistant (MDR) staphylococci was observed in 64.4% of the dogs, and approximately 39% had methicillin-resistant Staphylococcus sp. (MRS), of which 21.6% had MRSP and 1.9% had methicillin-resistant S. aureus (MRSA). The acquisition of MRSP during ICU hospitalization was associated with sex (female), age (>7 years), and dogs that had previously been treated with antimicrobials. Animals colonized by MRSP resistant to ≥9 antimicrobial classes had longer hospital stays than those colonized by other MRS strains. Among the 13 MRSP isolates that were subjected to whole-genome sequencing, ten were classified as ST71. A single nucleotide polymorphism (SNP) analysis revealed three clones, including one that was detected in infected dogs outside the ICU. This study indicates novel risk factors associated with colonization by MRSP. The detection of the same MRSP clone causing HAI outside the ICU reinforces the need for improved infection prevention and control practices at veterinary hospitals in general and at the ICU in particular.