scholarly journals Molecular Confirmation of Vancomycin-Resistant Staphylococcus aureus with vanA Gene from a Hospital in Kathmandu

2021 ◽  
Vol 2021 ◽  
pp. 1-8
Author(s):  
Meera Maharjan ◽  
Anil Kumar Sah ◽  
Susil Pyakurel ◽  
Sabita Thapa ◽  
Susan Maharjan ◽  
...  

Staphylococcus aureus, a commensal on the skin and in the nasal cavity of humans, is one of the most serious cases of nosocomial infections. Moreover, methicillin-resistant S. aureus (MRSA) is a leading cause of morbidity and mortality worldwide. For the treatment of MRSA infections, vancomycin is considered as a drug of choice. However, the emergence of vancomycin resistance among MRSA isolates has been perceived as a formidable threat in therapeutic management. To estimate the rate of vancomycin-resistant S. aureus (VRSA) and to detect the vancomycin-resistant genes, namely, vanA and vanB, among the isolates, a hospital-based cross-sectional study was conducted from July to December 2018 in Annapurna Neurological Institute and Allied Science, Kathmandu, Nepal. S. aureus was isolated and identified from different clinical samples and processed for antibiotic susceptibility testing by the modified Kirby–Bauer disc diffusion method. The screening of MRSA was performed as per Clinical and Laboratory Standard Institute (CLSI) guidelines. VRSA was confirmed by the minimum inhibitory concentration (MIC) method by employing E-test strips. All the phenotypically confirmed VRSA were further processed to detect the vanA and vanB gene by using the conventional polymerase chain reaction (PCR) method. A total of 74 (20.3%) S. aureus were isolated, and the highest percentage of S. aureus was from the wound samples (36.5%). Of 74 S. aureus isolates, the highest number (89.2%) was resistant to penicillin, and on the other hand, linezolid was found to be an effective drug. Likewise, 45 (60.81%) were found to be MRSA, five (11.11%) were VRSA, and 93.2% of S. aureus isolates showed an MAR index greater than 0.2. Two VRSA isolates (40%) were positive for the vanA gene. The higher prevalence of MRSA and significant rate of VRSA in this study recommend routine surveillance for the MRSA and VRSA in hospital settings before empirical therapy.

2020 ◽  
Vol 13 ◽  
pp. 117863372095207
Author(s):  
Kiran Duwadi ◽  
Sujan Khadka ◽  
Sanjib Adhikari ◽  
Sanjeep Sapkota ◽  
Pabitra Shrestha

Introduction: Patients with malignancies frequently develop infections as a result of surgical procedures and fungating wounds leading to pus formation. This cross-sectional study was conducted to explore the bacteriological spectra of infections of various cancer sites and their antibiotic sensitivity patterns among the patients visiting minor operation theatre (OT) of B.P. Koirala Memorial Cancer Hospital (BPKMCH), Chitwan, Nepal. Methods: Over a period of 3 months from September to November 2018, a total of 183 wound exudates and pus samples were collected and analyzed by standard microbiological procedures. Isolates were identified based on the colony characters, Gram staining and an array of biochemical tests. Antibiotic susceptibility testing was performed by Kirby-Bauer disc diffusion technique according to criteria set by CLSI, 2016. Methicillin resistance in Staphylococcus aureus was tested with the help of cefoxitin using disc diffusion method. Results: Out of the 183 samples, 149 (81.4%) were culture positive. Among 13 different isolates identified, S. aureus (43.0%) was predominant followed by E. coli (14.0%). Higher incidence of bacteria was seen among the males (52.3%), in the age group 51 to 60 years (26.8%) and among the patients undergoing surgical intervention to deal with cancer (34.2%). The prevalence of wound infection was significantly affected by gender, age, and treatment regimen ( P < .01). Out of the total 68 S. aureus isolates, 38 (44.1%) were deemed as Methicillin-resistant Staphylococcus aureus (MRSA). Among the 158 isolates, 85 (53.8%) were multi-drug resistant (MDR). Cefepime was the most effective antibiotic for Gram positive isolates whereas both imipenem and meropenem were found to be equally more effective for Gram negative isolates. Conclusion: This study suggests that patients with malignancies harbor pathogenic bacteria; therefore, prudent use of antibiotics is essential to prevent the emergence of MDR pathogens.


Author(s):  
Fibhaa Syed ◽  
Nasim Akhtar ◽  
Mohammad Ali Arif ◽  
Adil Ramzan ◽  
Rauf Niazi ◽  
...  

Abstract Objective: To determine the nasal carriage of staphylococcus aureus and methicillin-resistant staphylococcus aureus among healthcare workers in a tertiary care setting. Methods: The cross-sectional study was conducted at the Pakistan Institute of Medical Sciences, Islamabad, Pakistan, from April to July 2018, and comprised healthcare workers at the institution. Nasal swabs were collected and cultured on Mannitol salt agar. Mannitol fermenting colonies which were gram-positive cocci, catalase-positive and coagulase-positive were identified as staphylococcus aureus. Antibiotic susceptibility test was performed by modified Kirby-Bauer disc diffusion method. Methicillin resistance was detected using cefoxitin disc diffusion method. Data was analysed using SPSS 23. Results: Of the 210 nasal swabs, 52(24.76%) had a staphylococcus aureus growth, and, of them, 15(7.1%) were methicillin-resistant. No association could be established with either any single category of healthcare worker or an inter-department variation (p>0.05). Likewise, there was no association with age, gender, duration of service, smoking, co-morbidities, use of antibiotics in the preceding six months, treating a patient with methicillin-resistant staphylococcus aureus in the preceding six months and hospitalisation in the preceding year (p>0.05). Conclusion: The frequency of nasal carriage of methicillin resistant staphylococcus aureus amongst healthcare workers was regardless of the nature of their professional engagement. Key Words: Methicillin resistant staphylococcus aureus, Nasal carriage, Continuous...


2018 ◽  
Vol 24 (1) ◽  
pp. 625-631 ◽  
Author(s):  
Sahar Naz ◽  
Farhan Rasheed ◽  
Muhammad Saeed ◽  
Shagufta Iram ◽  
Ambereen Anwar Imran

Inter-hospital and intra-hospital dissemination of metallo-β-lactamase (MβL) producing strains possess significant therapeutic challenges. Objective: This study was carried out to evaluate the efficacy of Colistin against MβL producers. Material and Methods: This cross-sectional study was conducted in Microbiology Laboratory, Allama Iqbal Medical College, Lahore, Pakistan from 1stJuly 2016 to 25th February 2017. A total of 12126 clinical samples were collected from patients presenting to Jinnah Hospital, Lahore. Every sample was processed for bacterial culture. Bacterial identification was performed according to standard guidelines. Every gram-negative isolate was further processed for antimicrobial susceptibility testing by modified Kirby Baur disc diffusion method. Zone sizes were interpreted according to CLSI 2016 guidelines. Next day every carbapenem-resistant isolate were further processed for MβL detection by EDTA method, zone size of Carbapenem disc only and Carbapenem disc impregnated with EDTA was compared ( >7 mm increase MβL positive, 0-5 mm increase MβL-negative). Results: Out of total 12126 samples, 35.9% (n=4361) were culture positive and only 40.5% (n=1770) were Gram negative rods. Of these 9.6% (n=170) were Carbapenem-resistant isolates with 47% (n=80) MβL producers. Briefly 51.7% (n=30) Acinetobacter species were MβL positive, Pseudomonas species 38.5% (n=22), Escherichia coli 69.5% (n=16), Klebsiella species 37.0% (n=10), Proteus 66.6% (n=2) and 0% Citrobacter sppwere MβL positive. 32.5% MβL positive isolates were from ICU, 21.2% were from OPD, 12.5%were from Surgical Units, 12.5% were from Medical Unit, 17.5% were from Orthopedic Unit, and 3.7% were from Pulmonology ward. Almost 100% resistant was observed in MβL positive isolates for Imipenem,Piperacillin+ Tazobactum, Ceftriaxone, Co-amoxyclav, Cefoperazone+Sulbactam, Ciprofloxacin, and Amikacin, Doxycycline, and Gentamicin showed 91.2%, 94.0%, and 97.5% resistant rate respectively. No resistance was observed against Colistin. Conclusion: MβL producing Gram negative rods are rising in clinical setups. They are becoming a nightmare for clinicians to treat such infections. Colistin remains the only choice of drug for MβL positive and Negative isolates with 0% resistant rate except for Proteus species, to which it is intrinsically resistant.


2021 ◽  
Author(s):  
Mahesh Kumar Chaudhary ◽  
Indrani Jadhav ◽  
Megha Raj Banjara

Abstract BackgroundExtended spectrum β-lactamases are the group of beta-lactamase enzymes which confer resistance to the oxyimino-cephalosporins and monobactams. Emergence of ESBL producing genes which possess a serious threat for the treatment of infections both in community and hospitals since it is found to be increasing trends of multidrug resistance. This study was focused to find out the ESBLs producing genes.MethodsThis was a cross-sectional study conducted over a period of 2 years (September 2018 to April 2020) at microbiology laboratory of Nepal Mediciti Hospital. Clinical samples were processed in microbiology laboratory and culture isolates were indetified and characterized by standard microbiological techniques. following standard procedures.Antibiotic susceptibility testing was performed by modified Kirby-Bauer disc diffusion method as recommended by Clinical and Laboratory Standard Institute. Extended spectrum beta-lactamases were phenotypically confirmed by combined disc method.ESBL producing genes i.e. blaTEM,blaCTX-M and blaSHV were confirmed by PCR.ResultsOf the 1449 total E.coli isolates, 323/1449(22.29%) isolates were multi -drug resistance.Among total MDR Escherichia coli isolates, 215/323(66.56%) isolates were ESBL producers. The maximum number of ESBL Escherichia coli was isolated from urine 194(90.23%), followed by sputum 12(5.58%), swab 5 (2.32%), pus 2 (0.93%) and blood 2 (0.93%).Antibiotic susceptibility pattern of ESBL E.coli producers showed highest sensitivity towards tigecycline (100%) followed by polymyxin b, colistin and meropenem.Out of 215 phenotypically confirmed ESBL E.coli,only 186(86.51%) isolates were found to positive by PCR.The last 29(13.49%) were negative for any of the resistant genes.Among the ESBL genotypes,most common was blaTEM 118(63.4%) followed by blaCTX-M 68(36.6%).ConclusionThe emergence of MDR and ESBL producing E.coli isolates with high antibiotic resistant rates to commonly used antibiotics and increased predominance of major gene types blaTEM is a serious concern to the clinicians as well as microbiologist. This study forwarded a real massage to all the clinicians for the emergence of XDR and PDR resistant bacteria and preservation of antibiotics for their proper use in near future, if past experience with MDR and ESBLs is any indicator.


2021 ◽  
Vol In Press (In Press) ◽  
Author(s):  
Mohammad Hossein Zamanian ◽  
Marya Shirvani ◽  
Alireza Janbakhsh ◽  
Babak Sayad ◽  
Siavash Vaziri ◽  
...  

Background: Staphylococcus aureus is an opportunistic pathogen, which often causes asymptomatic diseases. This bacterium could cause several disorders, ranging from skin infections to life-threatening diseases. S. aureus could also develop resistance to a wide range of antibiotics. Objectives: The present study aimed to investigate the prevalence and pattern of antibiotic resistance in isolated S. aureus in Imam Reza Hospital of Kermanshah, Iran. Methods: This descriptive cross-sectional study was conducted on 2,228 patients with a positive S. aureus culture who were admitted to Imam Reza Hospital during 2016 - 2018. In accordance with the CLSI protocol, bacterial isolates were separated using laboratory tests, and antibiotic susceptibility was assessed using the standard disk-diffusion method. Data analysis was performed in SPSS version 24. Results: Most of the S. aureus-positive cases were isolated from the emergency ward (43.7%), blood samples (40.1%), and urine samples (23.9%). The highest antibiotic resistance was observed against erythromycin, penicillin G, ofloxacin, cefoxitin, clindamycin, and piperacillin. With 93.3% and 81.8% sensitivity, vancomycin and teicoplanin were respectively the most effective antibiotics against S. aureus. Conclusions: According to the results, the prevalence of resistant S. aureus strains could be due to the long-term hospitalization of patients and the overuse of antibiotics in infection treatment. Therefore, proper monitoring and development of effective infection control methods are essential in these healthcare settings.


2021 ◽  
Vol 59 (243) ◽  
pp. 1111-1115
Author(s):  
Shanti Shanti ◽  
Sanjib Mani Regmi ◽  
Nabina Shrestha

Introduction: Staphylococcus aureus, a superbug, resistant to multiple antibiotics led to growing interest in the usage of macrolide-lincosamide-streptogramin B antibiotics, which are now rapidly developing resistance. This study aims to find the prevalence of inducible clindamycin-resistant Staphylococcus aureus among obtained clinical samples from in-patient and out-patient departments of a tertiary care center. Methods: This is a descriptive cross-sectional study done in clinical samples from the in-patient and out-patient departments of a tertiary care center from September 2020-May 2021. Ethical clearance was taken from the Institutional Review Committee (Ref: 068/2077/2078). Staphylococcus aureus were isolated and antibiotic susceptibility tests were performed by disc diffusion method. Inducible clindamycin and methicillin resistance Staphylococcus aureus were detected using D-test and cefoxitin disc according to Clinical and Laboratory Standards Institute guidelines. Convenient sampling was done and the data was analyzed using Statistical Package for Social Sciences version 20. Point estimate at 95% confidence interval was calculated along with frequency and proportion for binary data. Results: Among a total of 141 Staphylococcus aureus isolated, the prevalence of inducible clindamycin resistant phenotype was 41 (29.1%) (21.6-36.59 at 95% Confidence Interval). Whereas, 30 (21.3%) were constitutive clindamycin resistant. The inducible 28 (47.5%) and 19 (32.2%) constitutive clindamycin resistance was higher among methicillin-resistant Staphylococcus aureus. Conclusions: The frequency of inducible clindamycin resistance among methicillin resistant Staphylococcus aureus was high, which alarms the use of macrolide-lincosamide-streptogramin B antibiotics in Staphylococcus aureus infections. Hence, D-test should be performed to detect inducible clindamycin resistance in routine testing to prevent treatment failure.


2019 ◽  
Vol 21 (2) ◽  
pp. 57-63 ◽  
Author(s):  
Ali Abdolahi ◽  
Alireza Khodavandi

Background and aims: Staphylococcus aureus is considered as a frequent cause of infections. Therefore, antibacterial agents including β-lactam are normally used to treat these infections while the emergence of antibiotic resistance is one of the major clinical problems in this respect. Accordingly, combination antibacterial therapy is one way to alleviate this problem. As a result, the present study aimed to investigate the combined effects of vancomycin and methicillin on clinical isolates of methicillin-resistant S. aureus (MRSA) and methicillinsensitive S. aureus (MSSA). In addition, a series of follow-up studies were performed to evaluate the antibacterial activity. Methods: The current cross-sectional study was conducted on 40 hospitalized patients from various clinical samples such as pus/wound swabs, blood, urine and sputum during a 6-month period in 2017. To this end, the antibacterial activities of vancomycin and methicillin, alone and in combination were investigated against MRSA and MSSA. Eventually, the inhibitory effects of vancomycin and methicillin alone and in combination were studied on the growth profile of MRSA and MSSA, as well as the expression of mecA gene. Results: Based on the results, the significant synergistic and partial synergistic activity with fractional inhibitory concentration (FIC) and fractional bactericidal concentration (FBC) indexes ranged from 0.27-0.938 and 0.313-0.844, respectively, in the combination of vancomycin and methicillin in MRSA and MSSA isolates. Further, the obtained data indicated that the combination of vancomycin and methicillin had a synergistic effect against MRSA isolates (P<0.01). The expression levels of the mecA gene were down-regulated by 5.25- fold in the combination of vancomycin and methicillin in MRSA isolate (P<0.001). Conclusion: In general, these events may reflect the potential uses of vancomycin and methicillin combination against MRSA. However, a greater understanding of the underlying molecular mechanisms of vancomycin and methicillin in combination could contribute to the development of therapeutic strategies.


Author(s):  
Ketaki Vyankatesh Kulkarni ◽  
Niranjan Pathak ◽  
Sandhya Sadanand Kulkarni

Introduction: Infections caused by Methicillin Resistant Staphylococcus aureus (MRSA) are associated with increased morbidity, longer antimicrobial therapy, etc. First option for treating invasive MRSA infections is glycopeptide vancomycin. Daptomycin, a lipopeptide rapidly bactericidal invitro against MRSA, is an acceptable alternative. Aim: To identify MRSA isolates from clinical specimens and assess their vancomycin and daptomycin susceptibility pattern. Materials and Methods: The cross-sectional study was conducted over a period of six months (January 2019 to June 2019) on 90 clinical samples in a rural teaching hospital in Pune, Maharashtra, India, including all samples except sputum received in the Microbiology laboratory. MRSA isolates were tested for vancomycin and daptomycin susceptibility by Epsilometer (E) test Minimum Inhibitory Concentration (MIC) method. Results: Among 90 MRSA isolates, most were from pus (51) followed by Urine (23), Blood (9), followed by Miscellaneous samples (7). Data analysis was done using Statistical Package for the Social Sciences (SPSS) version 16.0 software. All MRSA isolates in this study were susceptible to daptomycin with MIC in the range of 0.25-1 μg/mL with maximum isolates (39) with MIC of 0.38 μg/mL. Vancomycin MIC creep phenomenon was observed in 68 isolates. All these isolates also showed reduced susceptibility to daptomycin. Conclusion: MRSA in hospital set up mandates strict infection control practices in place. Daptomycin can be a good therapeutic alternative to treat infections caused by MRSA keeping in mind its therapeutic limitations and prior vancomycin usage in the same patient. Empirical therapy should always be based on antibiogram pattern. Adherence to hospital antibiotic policy and constant surveillance of antimicrobial resistance is the need of the hour.


2018 ◽  
Vol 9 (2) ◽  
pp. 281-286
Author(s):  
T. V. Sklyar ◽  
K. V. Lavrentievа ◽  
V. G. Gavrilyuk ◽  
N. V. Kurahina ◽  
M. O. Vereshchaha ◽  
...  

The therapy of infections, caused by methicillin-resistant Staphylococcus aureus (MRSA) with multiple resistance to antibiotics remains one of the most acute problems all over the world. It is all the more complicated since a priori the MSRA strains are not sensitive to the group of β-lactam antibiotics and multiresistant isolates are resistant to other groups of antimicrobial preparations, including antibiotics of choice (rifampicin, vancomycin, fusidic acid, co-trimoxazole and linezolid). From the samples of biomaterials of patients with pathological processes of different localization, we isolated 335 strains of bacteria, which were identified as Staphylococcus aureus, 169 (50.4%) of which were methicillin-resistant variants: 57.5% cultures were isolated from the nasal discharge; 50.7% – from faeces at intestinal dysbioses; by 40.0% – from conjunctival discharge, pharyngeal swab, outer ear swab and sputum; 33.3% – from urine samples. Antibiotic susceptibility of the isolated cultures was estimated by the disc-diffusion method and the method of serial dilution. The MRSA strains appeared to be most resistant to gentamycin, erythromycin (by 59.5% of cultures) and ciprofloxacin (53.3% of isolates), most sensitive – to vancomycin, co-trimoxazole and fusidic acid. The frequency of isolation of the cultures that are resistant to antibiotics did not exceed 4.1%. Rifampicin suppressed the growth of 75.8% and linezolid – of 100.0% of strains. Depending on the kind of biomaterial taken, MRSA strains, isolated from the nasal cavity, outer ear, urine samples, samples of sputum and faeces at intestinal dysbioses proved to be most resistant to the tested antimicrobial preparations. Rifampicin- and vancomycin-resistant strains of methicillin-resistant staphylococci made up 21.3% of the total number of the detected MRSA. They were most often isolated from the clinical samples taken from the nasal cavity and faeces. When determining minimal inhibitory concentration (MIC) of rifampicin and vancomycin, which are antibiotics of choice for treatment of infections caused by multiresistant MRSA, it was found that for 55.5% of the MRSA strains isolated from faeces, MIC of rifampicin coincided with the threshold value for this antibiotic and for 44.5%, it exceeded the threshold value by 2 times (4 µg/ml). 22.2% of them were characterized by the critical value of susceptibility to vancomycin (MIC ≥ 2 µg/ml). From rifampicin- and vancomycin-resistant MRSA stains, isolated from the nasal cavity, MIC of rifampicin coincided with the threshold value for this antibiotic for 66.7% of cultures, and exceeded it at least by 2 times for 33.3%. 11.1% of them were characterized by the critical level of susceptibility to vancomycin (MIC ≥ 2 µg/ml) and by 3.7% of strains exceeded MIC by 2 and 4 times respectively (4 and 8 µg/ml).


Author(s):  
Priyanka Pradhan Sneha Mohan ◽  
Tarana Sarwat Dalip Kakru

Methicillin resistant Staphylococcus aureus (MRSA) is major cause of nosocomial and community infections. Its prevalence varies with country and with hospitals within a country. Therefore, it is important for continuous surveillance in hospitals and other healthcare facilities in order to limit the spread of infections caused by MRSA. To determine the antibiogram of Staphylococcus aureus in a tertiary care hospital and to determine the change in trends in the antimicrobial susceptibility pattern of Staphylococcus aureus. To determine the prevalence of MRSA in a tertiary care hospital. This cross- sectional study was conducted at the Department of Microbiology, SMSR, Sharda University and Hospital. A total of 100 strains identified by morphological and biochemical characteristics were tested for antibiotic susceptibility using Kirby-Bauer disc diffusion method and the Prevalence of Inducible Clindamycin Resistance among the isolates. 100 out of 13,639 isolates clinical comprising samples were obtained Pus (78.6%), Blood (7%), Swab (4.1%), Sputum (4.1%), Urine (4.1%), Semen (2%). Maximum MRSA were obtained from pus samples (81%). However, out of total 7 isolates of Staphylococcus aureus obtained from blood 4 were MRSA and 3 were MSSA. The prevalence of MRSA that is (40%) infections was a high in our setup and is comparable to studies done earlier. This trend is particularly alarming for Staphylococcus aureus because of the severity and diversity of disease caused by this uniquely versatile pathogen.


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