Stocks of Neisseria were obtained from the Centers for Disease Control and Prevention and Food and Drug Association’s Antibiotic Resistance (AR) Isolate Bank “Neisseria species MALDI-TOF Verification panel.” Evolved strains included: AR-0944 (N. cinerea), AR-0945 (N. elongata), AR-0948 (N. canis), AR-0953 (N. subflava), and AR-0957 (N. subflava). Bacteria were cultivated for all subsequent protocols on GC agar base (Becton Dickinson Co., Franklin Lakes, NJ, USA) media plates containing 1% Kelloggs solution (GCP-K plates) for 18–24 hours at 37°C in a 5% CO2 atmosphere. Bacterial stocks were stored in trypticase soy broth (TSB) containing 20% glycerol at −80°C.
Gc agar base
Manufactured by BD
Sourced in United States
GC agar base is a microbiological culture medium used for the growth and isolation of Neisseria species, such as Neisseria gonorrhoeae. It provides the necessary nutrients and growth factors required for the cultivation of these fastidious microorganisms.
Automatically generated - may contain errors
Lab products found in correlation
Phosphate buffered saline (pbs), by Corning (1 mentions)
Yeast extract, by Thermo Fisher Scientific (1 mentions)
Tween 80, by Thermo Fisher Scientific (1 mentions)
Dextrose, by Thermo Fisher Scientific (1 mentions)
Brucella broth, by BD (1 mentions)
Isovitalex, by BD (1 mentions)
Protease peptone, by Thermo Fisher Scientific (1 mentions)
Chocolate 2 agar, by BD (1 mentions)
Pyridoxal, by Chem-Impex (1 mentions)
Auranofin, by Chem-Impex (1 mentions)
Azithromycin, by Tokyo Chemical Industry (1 mentions)
Ceftriaxone, by Tokyo Chemical Industry (1 mentions)
Hematin, by Chem-Impex (1 mentions)
Vitox, by Thermo Fisher Scientific (1 mentions)
4 protocols using gc agar base
1
Antibiotic-Resistant Neisseria Isolation
2
Diversity of Neisseria Antibiotic Resistance
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Stocks of Neisseria were obtained from the Centers for Disease Control and Prevention (CDC) and Food and Drug Association’s (FDA) Antibiotic Resistance (AR) Isolate Bank “Neisseria species MALDI-TOF Verification panel”. Evolved strains included: AR-0944 (N. cinerea), AR-0945 (N. elongata), AR-0948 (N. canis), AR-0953 (N. subflava), and AR-0957 (N. subflava). Bacteria were cultivated for all subsequent protocols on GC agar base (Becton Dickinson Co., Franklin Lakes, NJ, USA) media plates containing 1% Kelloggs solution (GCP-K plates) for 18–24 hours at 37°C in a 5% CO2 atmosphere. Bacterial stocks were stored in trypticase soy broth (TSB) containing 50% glycerol at −80°C.
3
Characterization of N. gonorrhoeae Strain FA1090
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The FA1090 (ATCC 700825) strain of N. gonorrhoeae was obtained from the American Type Culture Collection (ATCC) (Manassas, VA). GC agar base, Chocolate II agar, dried bovine hemoglobin, brucella broth and IsoVitaleX were obtained from Becton, Dickinson, and Company (Cockeysville, MD). Heart infusion agar was obtained from Hardy Diagnostics (Santa Maria, CA). Yeast extract and dextrose were obtained from Fisher Bioreagents (Fair Lawn, NJ). Auranofin, hematin, pyridoxal, and nicotinamide adenine dinucleotide (NAD) were obtained from Chem-Impex International (Wood Dale, IL). Protease peptone and VCNT supplement were purchased from Oxoid (Lenexa, KS). Phosphate-buffered saline (PBS) was obtained from Corning (Manassas, VA). Ceftriaxone, azithromycin and saponin were obtained from TCI America (Portland, OR) Tween 80 was obtained from Acros Organics (Fair Lawn, NJ). Estradiol pellets (5-mg, 21-day controlled-release) were purchased from Innovative Research of America (Sarasota, FL). Dacron swabs were purchased from the Medical Packaging Corporation (Camarillo, CA).
4
Characterizing Antimicrobial Resistance in N. gonorrhoeae
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A total of 94 N. gonorrhoeae isolates were selected: 58 were collected during 2012-2016 as part of the Gonococcal Resistance to Antimicrobials Surveillance Programme (GRASP); 8 17 were clinical isolates that had been referred to the PHE reference laboratory, generally owing to unusual resistance, and 19 were controls, including the 14 WHO reference strains. 9 The panel was selected to include isolates with a range of resistances variously to penicillin (chromosomally-mediated and plasmid-mediated), cefixime, ceftriaxone, azithromycin (moderately and highly raised MICs), ciprofloxacin, tetracycline (chromosomally-mediated and plasmid-mediated) and spectinomycin; it also included isolates that were fully susceptible to all of these antimicrobials.
All archived isolates were retrieved from -80C and inoculated on to non-selective GCVIT agar (GC agar base (Becton, Dickinson and Co, Le Pont de Claix, France) containing 1% Vitox (Oxoid, Basingstoke, UK)). Inoculated plates were incubated at 36C in 5% CO2 for 18-24 h. Growth was subcultured on to GCVIT agar plates and incubated again at 36C in 5% CO2 for 18-24 h. Identification of isolates as N. gonorrhoeae had previously been performed by real-time PCR using opa and porA targets, 10 or by MALDI-ToF.
All archived isolates were retrieved from -80C and inoculated on to non-selective GCVIT agar (GC agar base (Becton, Dickinson and Co, Le Pont de Claix, France) containing 1% Vitox (Oxoid, Basingstoke, UK)). Inoculated plates were incubated at 36C in 5% CO2 for 18-24 h. Growth was subcultured on to GCVIT agar plates and incubated again at 36C in 5% CO2 for 18-24 h. Identification of isolates as N. gonorrhoeae had previously been performed by real-time PCR using opa and porA targets, 10 or by MALDI-ToF.
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