A total of 386 Enterobacteriaceae isolates (188 Klebsiella pneumoniae, 122 Escherichia coli, 76 Enterobacter cloacae) were collected according to random selection within group (ESBL negative isolates were random collected from ESBL negative group, ESBL positive isolates were random collected from ESBL positive group, CRE isolates were random collected from CRE group) from two different regions of China (Chengdu and Chongqing), all microorganisms were isolated from patient specimens (such as Urine, blood, sputum and secretion) during treatment, and then those isolates have to be preserved for scientific research. All strains were identified by GN card (bioMerieux, Durham, NC, USA) and ID32 GN (bioMerieux, Durham, NC, USA).
Id32gn
Manufactured by bioMérieux
Sourced in France
The ID32GN is a biochemical identification system for Gram-negative bacteria. It provides rapid and reliable identification of a wide range of Gram-negative bacteria through a standardized and reproducible identification process.
Automatically generated - may contain errors
Lab products found in correlation
Id32strep, by bioMérieux (2 mentions)
Id32c strips, by bioMérieux (2 mentions)
Id32staph, by bioMérieux (2 mentions)
Gn card, by bioMérieux (1 mentions)
Cefotaxime, by Merck Group (1 mentions)
Meropenem, by Merck Group (1 mentions)
Ceftazidime, by Merck Group (1 mentions)
Macconkey agar, by Thermo Fisher Scientific (1 mentions)
Tryptic soy broth tsb, by Merck Group (1 mentions)
Orientation, by CHROMagar (1 mentions)
Id 32e, by bioMérieux (1 mentions)
Gram stain, by bioMérieux (1 mentions)
5 protocols using id32gn
1
Enterobacteriaceae Isolate Collection and Preservation
2
Bacterial and Fungal Identification in Semen
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In order to differentiate the bacterial species contaminating both the raw and diluted semen, each sample was plated onto Columbia agar with 5% (v/v) sheep blood and Mac-Conkey agar, respectively. The plates were incubated in aerobic conditions at 37 °C for 24 h. After this period, one colony of each morphotype was transferred onto tryptone soy agar and re-incubated for 24 h at 37 °C in order to obtain a fresh culture ready for identification. The identification of bacteria isolates was performed using Gram stain and ID32E, ID32GN, ID32STAPH, and ID32STREP (bioMérieux, Craponne, France). Filamentous fungi also called molds were identified on the basis of macroscopic and microscopic features using the primary cultures onto Sabouraud Chloramphenicol Agar. The yeasts were identified by biochemical tests using ID32C strips (bioMérieux, France) [41 ,42 ].
3
Isolation of Antibiotic-Resistant E. coli from Feces
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In total, 1 g of faeces was suspended in 10 ml of saline, and successive dilutions were performed of 1 : 10 and plated on MacConkey agar 100 µl of the dilution 1 : 100, 1 : 1000 and 1 : 10 000. When necessary to perform colony counting (between 30 and 150 c.f.u., approximately) the dilution 1 : 100 000 was plated. The dilution used for colony counts was plated in CHROMagar orientation (ChrOmagar). From the later medium, two E. coli
To search Gram-negative bacilli resistant to antibiotics, 100 µl of suspension in saline was plated on MacConkey agar (Oxoid, Hampshire, UK) with cefotaxime, ceftazidime or meropenem in subinhibitory concentrations (2 mg l−1) (Sigma-Aldrich, St Louis, MO, USA). At the same time, 1 g of faeces was suspended in 40 ml of Tryptic Soy Broth (TSB) (Sigma-Aldrich) and after incubation 200 µl were spread in the same culture media. Representative isolates were selected to confirm the identification using API 20E and ID32GN Biomérieux.
To search Gram-negative bacilli resistant to antibiotics, 100 µl of suspension in saline was plated on MacConkey agar (Oxoid, Hampshire, UK) with cefotaxime, ceftazidime or meropenem in subinhibitory concentrations (2 mg l−1) (Sigma-Aldrich, St Louis, MO, USA). At the same time, 1 g of faeces was suspended in 40 ml of Tryptic Soy Broth (TSB) (Sigma-Aldrich) and after incubation 200 µl were spread in the same culture media. Representative isolates were selected to confirm the identification using API 20E and ID32GN Biomérieux.
4
Bacterial and Fungal Identification in Semen
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The determinations were performed on semen samples collected at T0 and T2. To identify bacterial contamination, each sample was placed on the plate onto the Columbia agar with 5% (v/v) sheep blood and Mac-Conkey agar, respectively. The plates were incubated in aerobic conditions at 37 °C for 24 h. After this period, one colony of each morphotype was transferred onto the tryptone soy agar and re-incubated for 24 h at 37 °C in order to obtain a fresh culture ready for identification. The identification of the bacteria isolates was performed using Gram stain and ID32E, ID32GN, ID32STAPH, and ID32STREP (bioMérieux, Craponne, France). Filamentous levura (fungi/yeasts) were identified on the basis of macroscopic and microscopic features using the primary cultures onto a Sabouraud Chloramphenicol Agar. The yeasts were identified by biochemical tests using ID32C strips (bioMérieux, France) [22 ,54 ].
5
Isolation and Identification of Azithromycin-Resistant V. fluvialis
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Twenty-eight azithromycin-resistant V. fluvialis isolated from patients with cholera-like diarrhea admitted to Infections Diseases, Kolkata, between 2014 and 2015 were included in this study. V. fluvialis was isolated from stool specimens or rectal swabs in Cary-Blair medium using thiosulfate-citrate-bile salts-sucrose agar (TCBS), followed by overnight incubation at 37°C. Yellow colonies on the TCBS plates were examined by oxidase test, string test using 0.5% sodium deoxycholate solution, biochemical testing (ID 32GN; bioMérieux, Marcy l’Etoile, France) and by a species-specific V. fluvialis toxR PCR (4 (link)).
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