Author: Lee, Sung-Hwan; Kim, Ha-Young; Choi, Eun Wha; Kim, Doo
Title: Causative agents and epidemiology of diarrhea in Korean native calves Document date: 2019_10_25
ID: 3xf8fjxp_14
Snippet: Isolation of C. difficile was carried out according to a previous study [30] . The samples were mixed with the same amount of 100% alcohol, allowed to stand for 1 h at room temperature, subjected to alcohol shock, and centrifuged at 3,800 ×g for 10 min. The pellet was inoculated on C. difficile selective medium (CDSA, BD) and anaerobically cultured at 37°C for 48 h or more. Samples without pretreatment of alcohol shock were inoculated in approx.....
Document: Isolation of C. difficile was carried out according to a previous study [30] . The samples were mixed with the same amount of 100% alcohol, allowed to stand for 1 h at room temperature, subjected to alcohol shock, and centrifuged at 3,800 ×g for 10 min. The pellet was inoculated on C. difficile selective medium (CDSA, BD) and anaerobically cultured at 37°C for 48 h or more. Samples without pretreatment of alcohol shock were inoculated in approximately 1 mL of TCCFB (cycloserine-cefoxitin fructose broth containing 0.1% taurocholate sodium) and incubated at 37°C for 7 days. The suspected unique colonies on the selective medium were subcultured on a sheep blood agar, followed by purification of DNA. PCR was performed to confirm the presence of tcdB toxin gene in the purely isolated C. difficile.
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