Cefpodoxime

Manufactured by Mast Group

Sourced in United Kingdom

Cefpodoxime is a third-generation cephalosporin antibiotic used in laboratory settings. It is a semi-synthetic antibiotic with a broad spectrum of activity against various gram-positive and gram-negative bacteria.

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4 protocols using cefpodoxime

Phenotypic and Genotypic ESBL/AmpC Detection

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Two different ESBL/AmpC detection disc sets have been used to confirm ESBL/AmpC phenotypes. The first set is a combination of 4 individual discs of Cefotaxime/ Cefotaxime + Clavulanic acid/ Ceftazidime/ Ceftazidime + Clavulanic acid, purchased from either BD BBL TM or Oxoid company. The second set is a combination of 4 individual discs of Cefpodoxime/ Cefpodoxime + ESBL inhibitor/ Cefpodoxime + AmpC inhibitor/ Cefpodoxime + ESBL inhibitor + AmpC inhibitor, purchased from Mast Group company (D68C set).
In addition, AmpC and ESBL β-lactamase genes were detected using PCR assays. A total of three AmpC (bla CMY-2 , bla FOX , bla ACT-1/MIR-1 ) and ten ESBL (bla TEM , bla SHV bla CTX-M-1 , bla CTX-M-2 , bla CTX-M- positive isolates. Primers used in the PCR assays are listed in Table 6.
The MDR E. coli and Salmonella were selected for further experiments after being confirmed to exhibit ESBL/AmpC phenotypes.

Tran T., Checkley S., Caffrey N., Cassis R., Mainali C., Gow S., Agunos A., & Liljebjelke K. (2020). Genetic characterization of AmpC and extended-spectrum beta-lactamase (ESBL) phenotypes in Escherichia coli and Salmonella from Alberta poultry.

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Antibacterial Susceptibility Testing Protocol

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Antibacterial susceptibility test of isolates to cefepime (30 µg), cefotaxime (30 µg), cefoxitin (30 µg), ceftazidime (30 µg), ceftizoxime (30 µg), cefpodoxime (10 µg), imipenem (10 µg), meropenem (10 µg), ertapenem (10 µg), gentamicin (10 µg), amikacin (30 µg), ciprofloxacin (5 µg), and norfloxacin (10 µg) (Mast Group Ltd., Bootle, UK) was determined by disk diffusion method on Müller–Hinton agar media (Laboratorios CONDA, Madrid, Spain) according to the Clinical and Laboratory Standards Institute (CLSI).12 Minimum inhibitory concentration (MIC) of isolates to cefotaxime, cefepime, and imipenem was determined by microbroth dilution method according to CLSI. To determine MIC of colistin and tigecycline by microbroth dilution method, we used the European Committee on Antimicrobial Susceptibility Testing recommendations (http://www.eucast. org/clinical-breakpoints). Escherichia coli ATCC 25922 and Pseudomonas aeruginosa ATCC 27853 were used as standard strains in antibacterial susceptibility testing.

Kiaei S., Moradi M., Hosseini-Nave H., Ziasistani M, & Kalantar-Neyestanaki D. (2018). Endemic dissemination of different sequence types of carbapenem-resistant Klebsiella pneumoniae strains harboring blaNDM and 16S rRNA methylase genes in Kerman hospitals, Iran, from 2015 to 2017. Infection and Drug Resistance, 12, 45-54.

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ESBL-producing E. coli Identification

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Suspected ESBL-producing E. coli isolates (n = 1,140) were confirmed to be indole-positive using BBL Dry Slide (BD), and subsequently tested for ESBL-production by disk diffusion following CLSI guidelines [36 ], using Sensi-Discs (BD, Breda, the Netherlands). Zone diameters were determined for cefotaxime (30μg) ± clavulanic acid (10μg), ceftazidime (30μg) ± clavulanic acid (10μg), and cefoxitin (30 μg). ESBL-producing isolates were defined as strains resistant to cefotaxime (zone diameter ≤ 22 mm) and/or ceftazidime (zone diameter ≤ 17 mm), and an increase in zone diameter of ≥ 5 mm with the disks containing clavulanic acid [36 ]. ESBL-producing E. coli concentrations were calculated from the numbers of β-glucuronidase-positive colonies, and the fraction of isolates confirmed to be indole-positive and ESBL-producing. Some isolates (n = 22) did not appear to be resistant to either cefotaxime or ceftazidime, and then ESBL-production was confirmed using an alternative AmpC and ESBL detection test, which is based on cefpodoxime (Mastgroup Ltd., Bootle, UK). Third-generation cephalosporin-resistant isolates with no significant inhibitory effect of clavulanic acid (as defined by CLSI) were defined as non-ESBL-producers and excluded from further analyses (n = 32).

Blaak H., van Hoek A.H., Hamidjaja R.A., van der Plaats R.Q., Kerkhof-de Heer L., de Roda Husman A.M, & Schets F.M. (2015). Distribution, Numbers, and Diversity of ESBL-Producing E. coli in the Poultry Farm Environment. PLoS ONE, 10(8), e0135402.

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Phenotypic ESBL Detection in Enterobacterales

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All ESC-R isolates that generated characteristic ESBL-like colonies on the Brilliance ESBL Agar (n = 104) were subcultured onto SBA and further tested for demonstration of phenotypic ESBL-production with the combination disk test (CDT), including cefpodoxime, ceftazidime, and cefotaxime alone and in combination with clavulanic-acid (MAST Group, United Kingdom). In addition, all Enterobacterales selected on the Brilliance ESBL Agar were subject to further antimicrobial susceptibility testing by disk diffusion on Mueller-Hinton agar (MHA), according to the Clinical and Laboratory Standards Institute methodology [57 ]. The antimicrobial panel included: amoxicillin/clavulanic acid (30 μg), ampicillin (10 μg), aztreonam (30 μg), imipenem (10 μg), trimethoprim/sulfamethoxazole (25 µg), enrofloxacin (5 μg), tetracycline (30 μg), chloramphenicol (30 μg), and gentamicin (10 μg) (all disks and media were from Oxoid, Basingstoke, UK). E. coli ATCC 25,922 was used as the control for the disk diffusion susceptibility testing. Interpretation of the antimicrobial susceptibility results was done according to the CLSI [57 ]. An isolate was considered non-susceptible if it had intermediate or resistant results against the tested antimicrobial agent.

Cozma A.P., Rimbu C.M., Zendri F., Maciuca I.E, & Timofte D. (2022). Clonal Dissemination of Extended-Spectrum Cephalosporin-Resistant Enterobacterales between Dogs and Humans in Households and Animal Shelters of Romania. Antibiotics, 11(9), 1242.

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