Trimethoprim sulfamethoxazole

Manufactured by Mast Group

Sourced in United Kingdom

Trimethoprim-sulfamethoxazole is a combination antibiotic medication used for the treatment of various bacterial infections. It consists of two active ingredients, trimethoprim and sulfamethoxazole, which work together to inhibit the growth and reproduction of bacteria. This product is commonly used in laboratory settings for research and testing purposes.

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14 protocols using trimethoprim sulfamethoxazole

Antibiotic Susceptibility Test of CRAB

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The antibiotics susceptibility of isolates was determined using the disk diffusion method according to CLSI guidelines [17 ] and against the following antibiotics: imipenem (10 mg), ceftazidime (30 mg), ceftriaxone (30 mg), amikacin (30 mg), gentamicin (10 mg), tetracycline (30 mg), piperacillin/tazobactam (110 mg), ampicillin/sulbactam (20 mg), ciprofloxacin (5 mg), levofloxacin (5 mg) and trimethoprim/sulfamethoxazole (25 mg) (Mast Group Ltd, UK). The CRAB was defined when the isolate was resistant to imipenem [16 (link)].

Abdollahi A., Aliramezani A., Salehi M., Norouzi Shadehi M., Ghourchian S, & Douraghi M. (2021). Co-infection of ST2IP carbapenem-resistant Acinetobacter baumannii with SARS-CoV-2 in the patients admitted to a Tehran tertiary referral hospital. BMC Infectious Diseases, 21, 927.

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Methicillin Resistance Screening in S. aureus

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The disk diffusion method using cefoxitin (30 μg) disk in Mueller-Hinton agar (Merck, Germany) according to the CLSI was applied for the screening of methicillin resistance isolates. In addition, PCR assay was used for the detection of mecA gene.12
The Kirby–Bauer disk diffusion method was used to determine the susceptibility of the isolates against penicillin, ceftriaxone, amikacin, gentamicin, tobramycin, kanamycin, tetracycline, linezolid, teicoplanin, ciprofloxacin, rifampicin, quinupristin-dalfopristin, and trimethoprim-sulfamethoxazole (Mast Co., UK) based on the CLSI recommendation (CLSI 2019). The minimum inhibitory concentration (MIC) value for vancomycin, mupirocin, tigecycline, and fusidic acid was determined using the broth microdilution method. Results for fusidic acid and tigecycline were interpreted according to the European Committee for antimicrobial susceptibility testing (EUCAST) breakpoints (

http://www.eucast.org

). Low-level and high-level mupirocin resistance (LLMUPR, HLMUPR) were defined if MIC values of 8–256 µg/mL and ≥512 µg/mL were obtained. S. aureus strains ATCC 25923, ATCC 43300 and ATCC 29213 were used as reference strains.

Goudarzi M., Tayebi Z., Fazeli M., Miri M, & Nasiri M.J. (2020). Molecular Characterization, Drug Resistance and Virulence Analysis of Constitutive and Inducible Clindamycin Resistance Staphylococcus aureus Strains Recovered from Clinical Samples, Tehran – Iran. Infection and Drug Resistance, 13, 1155-1162.

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Methicillin-Resistant Staphylococcus aureus Profiling

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The methicillin resistance of 40 S. aureus isolates were tested by using the Oxacillin agar screen method on the Muller Hinton agar modified by adding NaCl (4 %) + Oxacillin (6 µg/ml) (Dhanalakshmi et al. 2012 ). The strains that could grow on this medium were then tested to be revealed whether they carry the mecA gene (Kot et al. 2020 (link); Ubukata et al. 1989 (link)), as described in the next section.
The disc diffusion method was conducted to determine the antibiotic resistance pattern of the identified methicillin-resistant S. aureus strains to Vancomycin (30 µg), Ciprofloxacin (5 µg), Erythromycin (15 µg), Tetracycline (30 µg), Gentamycin (10 µg), Ceftriaxone (30 µg), Amikacin (30 µg), Mupirocin (5 µg), Oxacillin (30 µg), Chloramphenicol (30 µg) and Trimethoprim/Sulfamethoxazole (1.25/23.75 µg) purchased from MAST Group, Merseyside, UK.

Khaleghi M, & Khorrami S. (2021). Down-regulation of biofilm-associated genes in mecA-positive methicillin-resistant S. aureus treated with M. communis extract and its antibacterial activity. AMB Express, 11, 85.

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Evaluating Antibiotic Resistance Patterns of MRSA

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The antibiotic resistance pattern of isolates was evaluated using the Kirby Bauer method according to the CLSI instructions against amikacin (AMK), clindamycin (CLI), ciprofloxacin (CIP), erythromycin (ERY), gentamicin (GEN), kanamycin (KAN), linezolid (LIN), penicillin (PEN), quinupristin-dalfopristin (SYN), rifampicin (RIF), tobramycin (TOB), tetracycline (TET), teicoplanin (TEC), and trimethoprim-sulfamethoxazole (SXT) (Mast Co., UK). MRSA strains were identified phenotypically using the Cefoxitin disk diffusion method (30 μg) according to the CLSI guidelines and detection of the mecA gene as previously described [4 (link)].
The microdilution was performed to determine minimum inhibitory concentration (MIC) titer for antibiotics vancomycin (VAN), tigecycline (TIG), and fusidic acid (FUS) according to the procedure detailed in our previous report [4 (link)]. Results for fusidic acid and tigecycline were interpreted based on the European Committee for antimicrobial susceptibility testing (EUCAST) recommendations (http://www.eucast.org). The inducible and constitutive macrolide-lincosamide-streptogramin group B (iMLS_B and cMLS_B) resistance phenotypes were identified by erythromycin and clindamycin disks by the CLSI guideline (CLSI 2019). S. aureus ATCC 25923, ATCC 43300, and ATCC 29213 strains were used for antibiotic susceptibility testing quality control.

Zamani S., Mohammadi A., Hajikhani B., Abiri P., Fazeli M., Nasiri M.J., Dadashi M., Goudarzi M, & Haghighi M. (2022). Mupirocin-Resistant Staphylococcus aureus in Iran: A Biofilm Production and Genetic Characteristics. BioMed Research International, 2022, 7408029.

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Antibiotic Susceptibility Testing Protocol

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Antibiotic susceptibility testing was performed by using Kirby–Bauer disc diffusion method according to Clinical and Laboratory Standards Institute recommendations (CLSI) [8 ]. The antimicrobial disks tested were ampicillin (AMP), cefoxitin (FOX), cefazolin (CFZ), ceftriaxone (CRO), cefotaxime (CTX), ceftazidime (CAZ), cefepime (FEP), amoxicillin-clavulanate (AMC), aztreonam (ATM), gentamicin (GEN), streptomycin (STR), amikacin (AMK), nalidixic acid (NAL), ofloxacin (OFX), ciprofloxacin (CIP), levofloxacin (LVX), chloramphenicol (CHL), trimethoprim-sulfamethoxazole (TMP/SMX), tetracycline (TET), azithromycin (AZM), imipenem (IPM) and meropenem (MEM) (Mast Co., Bootle, Merseyside, UK). Escherichia coli ATCC 25922 was used as the quality control strain. The MDR was defined as resistance to 3 or more unrelated antibiotics [9 ].

Zamanlou S., Rezaee M.A., Aghazadeh M., Ghotaslou R., Nave H.H, & Khalili Y. (2018). Genotypic Diversity of Multidrug Resistant Shigella species from Iran. Infection & Chemotherapy, 50(1), 29-37.

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Comprehensive Antibiotic Susceptibility Testing

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The Kirby–Bauer disc-diffusion method on Mueller–Hinton agar was used to test the susceptibility of the isolates against amikacin, gentamicin, tobramycin, kanamycin, tetracycline, erythromycin, clindamycin, linezolid, teicoplanin, ciprofloxacin, rifampicin, quinupristin-dalfopristin and trimethoprim-sulfamethoxazole (Mast Co., Bootle, UK) based on the CLSI guideline. Susceptibility to vancomycin, mupirocin, tigecycline and fusidic acid was assessed by the broth microdilution method to determine the MIC titre. The European Committee for Antimicrobial Susceptibility Testing (EUCAST) breakpoints was used to determine MIC titres of fusidic acid and tigecycline (EUCAST 2018). The results of other antibiotics were interpreted by using the CLSI 2018 breakpoints. Low-level and high-level mupirocin resistance (LLMUPR, HLMUPR), inducible macrolide-lincosamide-streptogramin group B (iMLS_B) and constitutive (cMLS_B) macrolide-lincosamide-streptogramin group B were identified based on the CLSI guideline. Susceptibility testing was quality controlled using S. aureus ATCC 25923, ATCC 43300 and ATCC 29213 strains. Powders of antibiotics were all obtained from Sigma Chemical Co. (St Louis, MO, USA).

Goudarzi M., Razeghi M., Chirani A.S., Fazeli M., Tayebi Z, & Pouriran R. (2020). Characteristics of methicillin-resistant Staphylococcus aureus carrying the toxic shock syndrome toxin gene: high prevalence of clonal complex 22 strains and the emergence of new spa types t223 and t605 in Iran. New Microbes and New Infections, 36, 100695.

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Genetic Determinants of Antimicrobial Resistance

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Genetic determinants for AMR were identified using staramr v0.5.1 (https://github.com/phac-nml/staramr) against the ResFinder^{78 (link)} and PointFinder^{79 (link)} databases. Phenotypic antimicrobial susceptibility testing was performed using the EUCAST disk diffusion method^{80 (link)} (Antibiotic disks; ampicillin 10 μg, chloramphenicol 30 μg and trimethoprim/sulfamethoxazole 25 μg from Mast Group). To compare the results of genotypic against phenotypic testing, sensitivity and specificity were calculated for first line agents used over the study period using MedCalc’s test evaluation calculator with Clopper-Pearson confidence intervals (https://www.medcalc.org/calc/diagnostic_test.php).

Pulford C.V., Perez-Sepulveda B.M., Canals R., Bevington J.A., Bengtsson R.J., Wenner N., Rodwell E.V., Kumwenda B., Zhu X., Bennett R.J., Stenhouse G.E., Malaka De Silva P., Webster H.J., Bengoechea J.A., Dumigan A., Tran-Dien A., Prakash R., Banda H.C., Alufandika L., Mautanga M.P., Bowers-Barnard A., Beliavskaia A.Y., Predeus A.V., Rowe W.P., Darby A.C., Hall N., Weill F.X., Gordon M.A., Feasey N.A., Baker K.S, & Hinton J.C. (2020). Stepwise evolution of Salmonella Typhimurium ST313 causing bloodstream infection in Africa. Nature Microbiology, 6(3), 327-338.

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Antibiotic Susceptibility Profiling of Bacterial Isolates

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The antibiotic susceptibility pattern of the isolates was determined by the disk agar diffusion method on Muller Hinton agar (Merck, Germany) according to the Clinical and Laboratory Standards Institute (CLSI) guidelines²¹. The antibiotics included piperacillin (100 µg), piperacillin-tazobactam (100/10 µg), imipenem (10 µg), meropenem (10 µg), doripenem (10 µg), ciprofloxacin (5 µg), levofloxacin (5 µg), trimethoprim-sulfamethoxazole (1.25-23.75 µg), ceftazidime (30 µg), cefotaxime (30 µg), and cefepime (30 µg) (MAST Co., England). The susceptibility pattern of the isolates against aminoglycosides including kanamycin, amikacin, spectinomycin, netilmicin, gentamicin, streptomycin, and tobramycin was determined using the micro-broth dilution method according to the CLSI guidelines²¹. For interpretation of the minimum inhibitory concentration (MIC) values, we referred to the CLSI guidelines and previous studies¹^,²¹^,²². Escherichia coli ATCC 25922 and A. baumannii ATCC 19606 were used as control strains for antibiotic susceptibility testing.

Jouybari M.A., Ahanjan M., Mirzaei B, & Goli H.R. (2021). Role of aminoglycoside-modifying enzymes and 16S rRNA methylase (ArmA) in resistance of Acinetobacter baumannii clinical isolates against aminoglycosides. Revista da Sociedade Brasileira de Medicina Tropical, 54, e05992020.

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Antibiotic Susceptibility Profiling of Isolates

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The susceptibility of isolates to antibiotics was determined using disk diffusion according to CLSI guidelines. The antibiotics tested were oxacillin (1 μg), imipenem (10 μg), meropenem (10 μg), clindamycin (2 μg), linezolid (30 μg), vancomycin (5 μg), ampicillin (2 μg), ceftazidime (30 μg), cefotaxime (30 μg), ceftriaxone (30 μg), amikacin (30 μg), gentamicin (10 μg), tigecycline (15 μg), piperacillin (100 μg), piperacillin/tazobactam (110 μg), ampicillin/sulbactam (20 μg), ciprofloxacin (5 μg), levofloxacin (5 μg), trimethoprim/sulfamethoxazole (25 μg), and colistin sulphate (25 μg) (Mast Group Ltd., Bootle, UK).

Mohammadnejad E., Seifi A., Ghanei Gheshlagh R., Aliramezani A., Fattah Ghazi S., Salehi M., Dehghan Manshadi S.A, & Orandi A. (2023). Determine phenotypical patterns of resistance to antibiotics in COVID-19 patients with associated bacterial infection: largest medical center in Iran. Iranian Journal of Microbiology, 15(3), 336-342.

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Antibiotic Susceptibility of DAEC Isolates

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Antibiotic susceptibility testing of the DAEC isolates to ampicillin, ampicillin-sulbactam, cefotaxime, ceftriaxone, ceftazidime, imipenem, gentamicin, nalidixic acid, trimethoprim-sulfamethoxazole, moxifloxacin and chloramphenicol (Mast Group Ltd, Bootle, UK) was carried out on Müller–Hinton agar (Merck Co.) by disc diffusion method according to the Clinical and Laboratory Standards Institute. Escherichia coli ATCC25922 was used as the quality control strain.

Javadi K., Mohebi S., Motamedifar M, & Hadi N. (2020). Characterization and antibiotic resistance pattern of diffusely adherent Escherichia coli (DAEC), isolated from paediatric diarrhoea in Shiraz, southern Iran. New Microbes and New Infections, 38, 100780.

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