Screening of erm Gene of Inducible Clindamycin Resistant Staphylococcus aureus

2019 ◽  
Vol 24 (2) ◽  
pp. 39-43
Author(s):  
Roshan Timsina ◽  
Bivek Timalsina ◽  
Anjana Singh

 Antibiotic resistance exhibited by Staphylococcus aureus is a growing global concern. This work was undertaken to determine the prevalence rate of inducible clindamycin resistant S. aureus in nasal sample and detect ermB gene in the isolates with inducible clindamycin resistance. Nasal swabs were collected from the school children and cultured on Mannitol Salt Agar (MSA) and Blood Agar (BA) for observation of colony morphology. Gram staining and biochemical test (catalase, oxidase, O-F and coagulase) were performed for further identification of the bacteria. The Kirby-Bauer disc diffusion method using a cefoxitin disc (30 μg) was used to detect methicillin resistant S. aureus (MRSA). All, the MRSA isolates were tested for ermB gene by PCR amplification. Among 64 S. aureus isolates, 17 (26 %) were MRSA. The prevalence of Inducible clindamycin resistant S. aureus (iMLSB) isolates was 23.4 % in the S. aureus isolates. All the isolates of MRSA were resistant to penicillin, while 88.2 % were sensitive to gentamicin. The prevalence of ermB gene was 3.1 % in the total S. aureus isolates and 11.7 % MRSA showed the presence of this gene. Routinely performing a D-test in laboratory will guide the clinicians on the rationale use of clindamycin and improving hygienic practices can reduce the spread of inducible clindamycin resistance.

2020 ◽  
Vol 2020 ◽  
pp. 1-10 ◽  
Author(s):  
Charu Arjyal ◽  
Jyoti KC ◽  
Shreya Neupane

Methicillin-resistant Staphylococcus aureus (MRSA) infection in human beings and animals is concerning; it stands out as one of the leading agents causing nosocomial and community infections. Also, marginally increasing drug resistance in MRSA has limited therapeutic options. This study focuses on estimating the prevalence of MRSA in shrines, a place where human and animal interaction is frequent, sharing antibiotic-resistant bacteria, antibiotic-resistant genes, and diseases. A total of 120 environmental swabs were collected from targeted areas during the study period, March 2018 to May 2018. Staphylococcus aureus was identified by growth on mannitol salt agar (MSA), and MRSA by growth on mannitol salt agar containing 4 μg Oxacillin, Gram staining, and conventional biochemical test. Isolates of S. aureus were characterized by antibiotic susceptibility testing using the disc diffusion method. MRSA and methicillin-sensitive S. aureus (MSSA) proportion were 19% and 81%, respectively; a high rate of MRSA was observed in isolates from Thapathali (28.6%). MSSA isolates showed a high rate of resistance to erythromycin (64.7%). MRSA isolates were resistant to gentamicin (50%), cotrimoxazole (25%), erythromycin (50%), and ciprofloxacin (25%). The isolates were susceptible to linezolid (100%), clindamycin (100%), ciprofloxacin (75%), erythromycin (50%), tetracycline (100%), and cotrimoxazole (75%). Intermediate resistance was also found in gentamicin (50%). Of the 11 MSSA isolates that were erythromycin resistant and clindamycin sensitive, 6 (54.5%) showed the inducible clindamycin resistance (ICR) pattern and 2 MRSA isolates that were erythromycin resistant and clindamycin sensitive showed ICR pattern. Fifteen MSSA isolates were β-lactamase positive, whereas only two MRSA isolates showed β-lactamase production. There exists a minimal research work on infectious diseases that are shared between primates and animals. This study suggests the pervasiveness of MRSA/MSSA in the shrines, which may be a primary place for pathogen exchange between humans and primates.


Author(s):  
Valery Silvery Sonola ◽  
Gerald Misinzo ◽  
Mecky Isaac Matee

We conducted this study to investigate the isolation frequency and phenotypic antibiotic resistance pattern of Staphylococcus aureus isolated from rodents, chickens, humans, and household soils. Specimens were plated onto mannitol salt agar (Oxoid, Basingstoke, UK) and incubated aerobically at 37 °C for 24 h. Presumptive colonies of S. aureus were subjected to Gram staining, as well as catalase, deoxyribonuclease (DNAse), and coagulase tests for identification. Antibiotic susceptibility testing was performed by using the Kirby–Bauer disc diffusion method on Mueller–Hinton agar (Oxoid, Basingstoke, UK). The antibiotics tested were tetracycline (30 μg), erythromycin (15 μg), gentamicin (10 μg), ciprofloxacin (5 μg), clindamycin (2 μg), and amoxicillin-clavulanate (20 μg/10 μg). The S. aureus strain American Type Culture Collection (ATCC) 25,923 was used as the standard organism. We found that 483 out of 956 (50.2%) samples were positive for S. aureus. The isolation frequencies varied significantly between samples sources, being 52.1%, 66.5%, 74.3%, and 24.5%, respectively, in chickens, humans, rodents, and soil samples (p < 0.001). S. aureus isolates had high resistance against clindamycin (51.0%), erythromycin (50.9%), and tetracycline (62.5%). The overall prevalence of multidrug-resistant (MDR) S. aureus isolates was 30.2%, with 8.7% resistant to at least four different classes of antibiotics.


Author(s):  
Joel Manyahi ◽  
Sabrina J. Moyo ◽  
Said Aboud ◽  
Nina Langeland ◽  
Bjørn Blomberg

AbstractDifficult-to-treat infections caused by methicillin-resistant Staphylococcus aureus (MRSA) are of concern in people living with HIV infection as they are more vulnerable to infection. We aimed to identify molecular characteristics of MRSA colonizing newly diagnosed HIV-infected adults in Tanzania. Individuals newly diagnosed with HIV infection were recruited in Dar es Salaam, Tanzania, from April 2017 to May 2018, as part of the randomized clinical trial CoTrimResist (ClinicalTrials.gov identifier: NCT03087890). Nasal/nasopharyngeal isolates of Staphylococcus aureus were susceptibility tested by disk diffusion method, and cefoxitin-resistant isolates were characterized by short-reads whole genome sequencing. Four percent (22/537) of patients carried MRSA in the nose/nasopharynx. MRSA isolates were frequently resistant towards gentamicin (95%), ciprofloxacin (91%), and erythromycin (82%) but less often towards trimethoprim-sulfamethoxazole (9%). Seventy-three percent had inducible clindamycin resistance. Erythromycin-resistant isolates harbored ermC (15/18) and LmrS (3/18) resistance genes. Ciprofloxacin resistance was mediated by mutations of the quinolone resistance-determining region (QRDR) sequence in the gyrA (S84L) and parC (S80Y) genes. All isolates belonged to the CC8 and ST8-SCCmecIV MRSA clone. Ninety-five percent of the MRSA isolates were spa-type t1476, and one exhibited spa-type t064. All isolates were negative for Panton-Valentine leucocidin (PVL) and arginine catabolic mobile element (ACME) type 1. All ST8-SCCmecIV-spa-t1476 MRSA clones from Tanzania were unrelated to the globally successful USA300 clone. Carriage of ST8 MRSA (non-USA300) was common among newly diagnosed HIV-infected adults in Tanzania. Frequent co-resistance to non-beta lactam antibiotics limits therapeutic options when infection occurs.


2019 ◽  
Vol 6 ◽  
pp. 59-62
Author(s):  
Ranjana K.C. ◽  
Ganga Timilsina ◽  
Anjana Singh ◽  
Supriya Sharma

Objectives: To isolate methicillin resistant Staphylococcus aureus (MRSA) from anterior nares of dairy workers and dairy products and assess the antibiotic susceptibility pattern of the isolates. Methods: Swab samples collected from anterior nares of dairy workers and dairy product (butter) were inoculated into mannitol salt agar and incubated at 37ºC for 24 hours. Identification was done based on colony characteristics, Gram's staining, catalase, oxidase and coagulase test. Antibiotic susceptibility testing was done by modified Kirby Bauer disc diffusion method. MRSA was confirmed by using cefoxitin disc. Results: A total of 109 S. aureus (98 from dairy workers and 11 from butter samples) were isolated. Out of them 32 MRSA were isolated from dairy workers and 4 from butter samples. The association between age group and MRSA was found insignificant (p = 0.115). The association of MRSA between male and female workers was found significant (>0.05). About 86% of the MRSA isolates were susceptible to Gentamicin (86.11%) followed by Ciprofloxacin (77.78%). Conclusion: Detection of MRSA among dairy workers and dairy products warrants proper handling and adequate control measures to prevent transmission of MRSA from dairy industry.


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...


2013 ◽  
Vol 30 (1) ◽  
pp. 31-36 ◽  
Author(s):  
Marina Dinić ◽  
Slavica Vuković ◽  
Dobrila Stanković Đorđević ◽  
Milena Bogdanović

Summary Staphylococcus aureus (S. aureus) is a microorganism that colonizes the skin and mucosal surfaces of healthy individuals, but it is also one of the most common causes of community-acquired and hospital infections. Nasal carriage of S. aureus represents a major risk factor for the development of infection with this bacterium. A special therapeutic problem are methicillin-resistant isolates of S. aureus (MRSA). The aim of this study was to assess the nasal carriage of S. aureus in healthy individuals in the local community, and the sensitivity of the microorganism to antibiotics. The study enrolled 56.868 healthy individuals aged 19 to 65 years, and 2.040 healthy school children aged 15 to 19 years. Specimens to be studied were obtained from anterior nares. We used the disk diffusion method (Kirby-Bauer) on Mueller- Hinton agar to assess the sensitivity of isolated S. aureus. S. aureus was isolated in 1.381 (2.34%) respondents. Positive findings were obtained in 2.33% of adult examinees, and in 2.59% of studied school children. We found a low level of susceptibility only to penicillin (5.36%). The susceptibility of S. aureus isolates to all other tested antibiotics was present in a high percentage, with the lowest percentage of susceptibility to doxycycline (71.54%) and erythromycin (86.09%). The highest percentage of susceptibility of tested isolates was reported for fusidic acid (99.27%). In relation to the total number of S. aureus isolates from nasal swabs in adults, MRSA was present in 8.96% (119 isolates), while there were 4 MRSA isolates from nasal swabs in school children. In this study, we established a low percentage of nasal carriage of S. aureus in the population of healthy individuals, but a high percentage of MRSA.


2012 ◽  
Vol 4 (2) ◽  
pp. 68-78
Author(s):  
Lívia Cafundó Almeida ◽  
Marcus Vinicius Pimenta-Rodrigues ◽  
Daniela Vanessa Moris ◽  
Carlos M. Castelo Branco Fortaleza ◽  
Maria de Lourdes R.de Souza da Cunha

Staphylococcus aureuscan cause a variety of infections, especially nosocomial. Its importance lies in the combination of virulence, invasiveness and antibiotic resistance constituting therapeutic challenges. This study aimed to assess the genotype and phenotypic resistance profiles of samples of S. aureusisolated from hospitalized patients in a Brazilian teaching hospital. We evaluated 1078 samples obtained from surveillance cultures and clinical S.aureusin hospitalized patients. To assess the phenotypic resistance profile was used disk diffusion method according to CLSIcriteria, 2011. For the determination of genotypic resistance was the presence of the mecAgene by polymerase chain reaction -PCR. Of the 1078 samples tested phenotypically, it was observed that 75.1% of the samples were Methicillin Resistant Staphylococcus aureus(MRSA), and of these, 98.4% were resistant to oxacillin and 100%, cefoxitin. To determine genotype was performed for the PCR amplification of the mecAgene. Of the 443 samples tested for the mecA336 samples were positive (75.8%). Of these samples, 85.7% showed resistance phenotype oxacillin and cefoxitin 88.4%. Due to the high rate of MRSA, concludes the need for investment in research, rational use of antimicrobials and creation of reference laboratories for verification of antimicrobial resistance


2021 ◽  
Vol 7 (1) ◽  
pp. FSO361
Author(s):  
Roshan Timsina ◽  
Upasana Shrestha ◽  
Anjana Singh ◽  
Bivek Timalsina

Aim: Resistance to methicillin and Macrolide–Lincosamide and Streptogramins B and their association with erm genes in Staphylococcus aureus are unknown in Nepal. Materials & methods: Nonduplicate nasal swabs from 160 school children were collected from April to September 2018 and processed using standard microbiological procedures. Results: Out of 160 samples, 64 (40%) were S. aureus in which 17 (26.6%) were methicillin-resistance Staphylococcus aureus (MRSA). D-test identified 15 (23.4%) as inducible clindamycin-resistant, which were more prevalent in MRSA (76.4%) than methicillin-sensitive S. aureus (MSSA; 4.2%). 18.7% of isolates harbored the ermC gene followed by ermA (15.6%) and ermB (3.1%), and were more in MRSA than MSSA. Conclusion: To prevent treatment failure by inducible resistance, D-test must be performed on erythromycin-resistant and/or clindamycin-sensitive isolates.


2016 ◽  
Vol 5 (1) ◽  
pp. 1-5 ◽  
Author(s):  
Ganesh Kumar Singh ◽  
Bigu Kumar Chaudhari ◽  
Kamal Prasad Parajuli

Background Resistance to antimicrobial agents is prevalent among Staphylococci. This has led to wide uses of macrolide-lincosamide-streptogramin B (MLSB) antibiotics to Staphylococcus aureus (S. aureus) infections. MLSB though chemically distinct, have similar target site and mode of action. The multiple mechanisms are responsible for resistance to MLSB antibiotics which can lead to clinical failure. The aim of the study was to investigate the frequency of inducible and constitutive clindamycin resistance among clinical isolates of S. aureus and their relationship with Methicillin-resistant Staphylococcus aureus (MRSA).Material & Methods A total of 336 unique Staphylococcus aureus isolates from different clinical samples obtained from patients were studied. Antibiotic susceptibility test was performed by Kirby- Bauer disc diffusion method. “D test” was performed to detect inducible clindamycin resistance as per CLSI guidelines. MRSA was detected using Cefoxitin (30μg) and results were interpreted according to CLSI criteria.Results Inducible clindamycin resistance was seen in 45 (13.39%), constitutive clindamycin resistance was seen among 58 (17.26%) while MS phenotype was observed among 38(11.30%) of isolates. Inducible resistance as well as constitutive resistance was higher among MRSA as compared to MSSA (21.11%, 4.48% and 21.11%, 12.82%respectively).Conclusion The Successful use of clindamycin for the treatment of infection caused by S. aureus can be predicted based on the result of simple and inexpensive D test.Journal of Nobel Medical CollegeVolume 5, Number 1, Issue 8, January-July 2016, 1-5


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