Trimethoprim

Manufactured by Bio-Rad

Sourced in France

Trimethoprim is a laboratory reagent and antimicrobial agent used in research and diagnostic applications. It is a synthetic compound that inhibits the enzyme dihydrofolate reductase, which is essential for the synthesis of DNA and cell growth. Trimethoprim is commonly used in combination with other antimicrobial agents for various applications in the life sciences.

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Trimethoprim/sulfamethoxazole (14 citations) Trimethoprim-sulfamethoxazole (SXT) (2 citations) Trimethoprim-sulfametoxazole (2 citations)

5 protocols using trimethoprim

Evaluating Antibiotic Resistance in CTX-R E. coli

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The antibiotic susceptibilities of CTX-R E. coli and their transconjugants were determined by disk diffusion method according to CLSI protocols (19 ). The following antibiotic disks were used: ampicillin (10 μg), amoxicillin (20 μg) plus clavulanic acid (10 μg), cefoxitin (30 μg), ceftazidime (30 μg), cefotaxime (30 μg), cefepime (30 μg), aztreonam (30 μg), imipenem (10 μg), streptomycin (10 μg), gentamicin (10 μg), kanamycin (30 μg), nalidixic acid (30 μg), ciprofloxacin (5 μg), tetracycline (30 μg), chloramphenicol (30 μg), trimethoprim (5 μg), and sulfonamides (300 μg) (Bio-Rad, Marnes-la-Coquette, France if available or Oxoid, Dardilly, France). The susceptibility breakpoints for all antimicrobials were those recommended by CLSI (20 ).
ESBL production was detected by double-disk synergy test on Mueller-Hinton agar between clavulanic acid and ceftazidime, cefotaxime, or cefepime (20 , 21 (link)). AmpC beta-lactamases detection was based on the inhibitory effect of cloxacillin on AmpC production observed on plates supplemented with 200 mg/L cloxacillin. The control strains used were E. coli ATCC 25922, K. pneumoniae ATCC 700603, and K. pneumoniae CMY-2 from Pr R. Bonnet, France.

Dupouy V., Abdelli M., Moyano G., Arpaillange N., Bibbal D., Cadiergues M.C., Lopez-Pulin D., Sayah-Jeanne S., de Gunzburg J., Saint-Lu N., Gonzalez-Zorn B., Andremont A, & Bousquet-Mélou A. (2019). Prevalence of Beta-Lactam and Quinolone/Fluoroquinolone Resistance in Enterobacteriaceae From Dogs in France and Spain—Characterization of ESBL/pAmpC Isolates, Genes, and Conjugative Plasmids. Frontiers in Veterinary Science, 6, 279.

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Antimicrobial Susceptibility Testing of Enterobacteriaceae

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Disk diffusion method was used to test and confirm the antimicrobial susceptibility of the Enterobacteriaceae isolates using Muller–Hinton agar (MHA, Oxoid, Milan, Italy) and an incubation time of 16–18 h at 37 °C, following the Clinical and Laboratory Standards Institute Guidelines (CLSI) [24 ]. The antimicrobial used were: ciprofloxacin (CIP, 5 µg), nalidixic acid (NA, 30 µg), amoxicillin/clavulanic acid (AMC, 20/10 µg), amoxicillin (AML, 25 µg), levofloxacin (LEV, 5 µg), cefotaxime (CTX, 30 µg), sulphonamides (SSS, 300 µg), tetracycline (TE, 30 µg), trimethoprim/sulphamethoxazole (SXT, 1,25/23,75 µg), trimethoprim (TMP, 5 µg), chloramphenicol (C, 30 µg), and neomycin (N, 30 µg) (Bio-Rad, Marnes la Coquette, France). The results were assessed following the CLSI guidelines [24 ].

Lo Vecchio G., Cicero N., Nava V., Macrì A., Gervasi C., Capparucci F., Sciortino M., Avellone G., Benameur Q., Santini A, & Gervasi T. (2022). Chemical Characterization, Antibacterial Activity, and Embryo Acute Toxicity of Rhus coriaria L. Genotype from Sicily (Italy). Foods, 11(4), 538.

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Urine Culture and Susceptibility Testing

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Isolate identification and phenotypic susceptibility testing were performed according to standard procedures at each pathology service. At Dorevitch Pathology, the MSU submitted for laboratory testing was cultured on CHROMID CPS Elite (CPSE) agar incubated overnight at 35°C–37°C in air (bioMérieux). Identification of isolates was confirmed using MALDI-TOF MS on the VITEK MS platform (bioMérieux). Susceptibility testing was performed according to CLSI guidelines using disc diffusion against ampicillin, amoxicillin/clavulanate, cefazolin, ceftriaxone, ciprofloxacin, trimethoprim and nitrofurantoin (Bio-Rad) and supplementary susceptibility testing to fosfomycin, meropenem, ertapenem and gentamicin was performed in the event of E. coli ceftriaxone non-susceptibility.¹¹ Where susceptibility testing was not performed for Staphylococcus saprophyticus, we assumed trimethoprim susceptibility.^{12 (link)} We focused on E. coli for genomic characterization as it is the most frequently isolated bacterium in urine.

Curtis S.J., Kwong J.C., Chaung Y.L., Mazza D., Walsh C.J., Chua K.Y, & Stewardson A.J. (2023). Resistance to first-line antibiotic therapy among patients with uncomplicated acute cystitis in Melbourne, Australia: prevalence, predictors and clinical impact. JAC-Antimicrobial Resistance, 6(1), dlad145.

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Antibiotic Susceptibility Testing Protocol

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Antibiotic susceptibility was determined by disc diffusion on Mueller–Hinton agar, in accordance with the guidelines of the Antibiogram Committee of the French Society for Microbiology (CA-SFM & EUCAST, 2014 ). The following 32 antimicrobial drugs (Bio-Rad) were tested: ampicillin, ticarcillin, piperacillin, piperacillin/tazobactam, cefamandole, cefoperazone, cefoxitin, cefotaxime, amoxicillin/clavulanic acid, ticarcillin/clavulanic acid, imipenem, meropenem, ertapenem, cefepime, ceftazidime, streptomycin, spectinomycin, kanamycin, amikacin, gentamicin, netilmicin, tigecycline, isepamicin, nalidixic acid, pefloxacin, ciprofloxacin, sulfonamides, trimethoprim, sulfamethoxazole/trimethoprim, chloramphenicol, tetracycline and azithromycin. Minimal inhibitory concentration (MIC) values for nalidixic acid, ciprofloxacin and azithromycin were determined by Etests (bioMérieux). E. coli CIP 76.24 (ATCC 25922) was used as a control.

Kuijpers L.M., Le Hello S., Fawal N., Fabre L., Tourdjman M., Dufour M., Sar D., Kham C., Phe T., Vlieghe E., Bouchier C., Jacobs J, & Weill F.X. (2016). Genomic analysis of Salmonella enterica serotype Paratyphi A during an outbreak in Cambodia, 2013–2015. Microbial Genomics, 2(11), e000092.

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Antimicrobial Resistance Profiling of Diarrheagenic E. coli

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Thirty-one (31) diarrheagenic Eschericia coli (DEC) were subjected to the antimicrobial susceptibility testing. It was carried out by disk diffusion method on Müller-Hinton agar (Liofilchem, Italy) according to the recommendations of the European Committee on Antimicrobial Susceptibility Testing (EUCAST) [10 ]. After depositing the antibiotics, the plates were incubated at +37 °C for 18–24 h. The diameters of the antibiotic sensitivity halos were recorded according to the recommendations of EUCAST. Intermediary (I) susceptibility of pathovars was classified as resistant (R). According to EUCAST recommendations on antibiotics used in case of infection with enterobacteria and in view of the multidrug resistance observed in recent years, 19 antibiotics divided into 7 different families were tested. These include amoxicillin (25 µg), amoxicillin–clavulanic acid (20/10 µg), ceftriaxone (30 µg), cefotaxime (30 µg), cefepime (30 – g), cefixime (10 µg), piperacillin (75 µg), piperacillin–tazobactam (100 +10 µg), imipenem (10 µg), tetracycline (30 µg), chloramphenicol (30 µg), trimethoprim–sulfametoxazole (1.25 ± 23.75 µg), aztreonam (30 µg), colistin sulfate (50 µg), ciprofloxacin (5 µg), nalidixic acid (30 µg), gentamycin (15 µg), netilmicin (10 µg), and tobramycin (10 µg) (Bio-Rad, France).

Konaté A., Dembélé R., Guessennd N.K., Kouadio F.K., Kouadio I.K., Ouattara M.B., Kaboré W.A., Kagambèga A., Cissé H., Ibrahim H.B., Bagré T.S., Traoré A.S, & Barro N. (2017). Epidemiology and Antibiotic Resistance Phenotypes of Diarrheagenic Escherichia Coli Responsible for Infantile Gastroenteritis in Ouagadougou, Burkina Faso. European Journal of Microbiology & Immunology, 7(3), 168-175.

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