From December 2015 to September 2016, 196 non-repetitive isolates of K. pneumoniae were collected from the urine samples of patients with UTI who were referred to medical diagnostic laboratories in Yasuj, Southwestern Iran. Each urine sample was cultured on EMB and Blood Agar (Merck, Germany) and incubated at 37°C for 24 hr. Using conventional biochemical tests on culture media such as Methyl red-Voges Proskauer (MR-VP), Triple Suger İron (TSI), Sulfide İndole Motility (SIM), Simmons Citrate, and Urea Agar (Merck, Germany), identification of K. pneumoniae isolates was performed 19 . Verified isolates of K. pneumoniae were stored at −20°C in TSB medium (Merck, Germany) with 15% glycerol for next steps. This study was approved by the ethical committee of Yasuj University of Medical Sciences.
Urea agar
Manufactured by Merck Group
Sourced in Germany
Urea Agar is a microbiological culture medium used for the detection and identification of urea-splitting microorganisms. It is composed of urea, along with other essential nutrients and growth factors. The presence of urea in the agar allows for the identification of bacteria that can hydrolyze urea, a key characteristic of certain species.
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Lab products found in correlation
Xylose lysine deoxycholate agar, by Merck Group (2 mentions)
Tsb medium, by Merck Group (1 mentions)
Simmons citrate, by Merck Group (1 mentions)
Triple sugar iron agar, by Merck Group (1 mentions)
Eosin methylene blue agar, by Merck Group (1 mentions)
Simmons citrate agar, by Merck Group (1 mentions)
Tryptic soy broth (tsb), by Merck Group (1 mentions)
Triple sugar iron agar, by HiMedia (1 mentions)
Tetrathionate broth, by HiMedia (1 mentions)
Salmonella shigella agar, by BD (1 mentions)
Sim medium, by BD (1 mentions)
Simmons citrate agar, by HiMedia (1 mentions)
Lysine iron agar, by BD (1 mentions)
4 protocols using urea agar
1
Isolation and Identification of K. pneumoniae from UTI
2
Isolation and Identification of Shigella spp. from Pediatric Diarrhea
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We collected 3,254 stool culture of children <16 years of age with diarrheal infections referred to Golestan and Abuzar Hospitals during a period of 10 months from August 2016 to June 2017. The study design was approved by the Research Ethics Committee of Ahvaz Jundishapur University of Medical Sciences, Iran (IR.AJUMS.REC.1395.427). These samples were inoculated into Gram-negative (GN) Broth tubes as an enrichment medium as a part of the routine hospital laboratory procedure and then immediately transferred to the Laboratory of Microbiology Department of Medicine School of Ahvaz, Iran. From each patient, only one Shigella isolate in the diarrheal phase was included in this study. The GN Broth tubes were incubated at 37°C for 4–6 hrs and then streaked on Xylose Lysine Deoxycholate Agar and Eosin Methylene Blue Agar (Merck-Germany). All plates were incubated at 37°C for 24–48 hrs. The suspected colonies were biochemically identified as Shigellaspp. using Lysine Decarboxylase Agar, Sulfide-indole-motility, Triple-sugar iron Agar, Urea Agar and Simmons citrate Agar (Merck-Germany).11
3
Identification of Clinical Klebsiella pneumoniae
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This study was carried out at the Microbiology Laboratory of Shahid Beheshti University of Medical Sciences (Tehran, Iran). In total, 60 non-repetitive clinical K. pneumoniae isolates were taken from various clinical specimens existing in 9 hospitals in 4 provinces in Iran from 2019 to 2020. Further testing was employed to identify K. pneumoniae clinical isolates using biochemical tests such as oxidase test, the reaction on TSI culture medium, indole production and motility on SIM medium, the reaction in MR-VP medium, growth on Simmon-citrate medium, and urease production on urea agar (Merck Darmstadt, Germany) [17] . The isolates were maintained in Tryptic Soy Broth (TSB) (Merck, Germany) with 20% glycerol at À70 8C.
4
Isolation and Identification of Salmonella
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The samples for Salmonella isolation and identification were tested using the ISO 6579 protocol. After incubation, the pre‐enriched media (TSB) were cultured in a ratio of 1:10 on tetrathionate broth (HiMedia, LOT no. M032) and in a ratio of 1:100 on Rappaport Vassiliadis medium (HiMedia, LOT no. M880), followed by incubation at 37 and 42°C, respectively. After the incubation period, the cultures were streaked on xylose–lysine–deoxycholate agar (Merck, LOT no. 105287) and Salmonella‐Shigella agar (BD, LOT no. 274500) media. Suspected colonies for Salmonella spp. (H2S‐producing and non‐lactose fermenter colonies) were selected for confirmatory biochemical tests. Isolates were cultured on Triple Sugar–Iron agar (HiMedia, LOT no. 211825), SIM medium (BD, LOT no. 211578), urea agar (Merck, LOT no. 108492), lysine iron agar (BD, LOT no. 211363), Simmons citrate agar (HiMedia, LOT no. M099) and Methyl Red‐Voges Proskauer broth (HiMedia, LOT no. GM070) for further confirmation of Salmonella spp. ONPG disks (HiMedia, LOT no. DD008) were used to detect the beta‐galactosidase activity of late lactose fermenters (Bahramianfard et al., 2021 (link)).
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