Isolator 10 tube

Salmonella Typhi shedding in stool was assessed using daily self-collected samples, cultured according to national standard operating procedures [14 ]. Blood (10 mL or 5 mL at typhoid diagnosis) was cultured daily by direct inoculation into broth (BACTEC Plus Aerobic vials, BD) and subsequent automated growth detection (BACTEC FX System, BD), in accordance with standard methods [15 ]. Salmonella Typhi growth and serotype were confirmed by biochemical profile (API-10S, bioMérieux, France) or slide agglutination according to the Kauffman-White classification, respectively [16 ].
Quantitative blood culture was performed at typhoid diagnosis by inoculation of 10 mL blood into an ISOLATOR 10 tube (a commercial lysis-centrifugation system; Alere, UK). After centrifugation, the resulting pellet was directly plated onto XLD agar (Oxoid, UK). Quantitative stool culture was performed by suspending 1 g of stool in sodium selenite, followed by subculturing onto XLD agar (Oxoid). Blood- or stool-inoculated plates were then incubated (37°C for 24 hours) prior to identification and counting.

Daily clinical review and temperature measurements were carried out for 7 days after vaccination and 28 days after challenge. In addition, participants collected symptom data daily (solicited for headache, feeling generally unwell, loss of appetite, abdominal pain, nausea/vomiting, myalgia, arthralgia, cough, diarrhea and constipation, and any unsolicited symptoms) (14 (link)). Symptom severity scores were calculated by summing maximum severity values assigned to each of up to 10 symptoms experienced daily during 14 days after challenge and deriving the mean (14 (link)).
Blood (10 mL) samples were collected for bacterial culture at each visit. Cultures were performed by the local hospital accredited pathology laboratories according to national standard operating procedures (22 –24 ), and as previously described (14 (link), 15 (link)). Quantitative blood culture samples were collected immediately prior to antibiotic treatment by inoculation of 10-mL blood into an ISOLATOR 10 tube (Alere, UK).

Quantitative blood culture was performed by using Wampole ISOSTAT/ISOLATOR Microbial System. Venous blood (10 ml) was lyzed by adding directly into an ISOLATOR 10 tube (Alere, UK). Following blood lysis, the tube was centrifuged for 30 min at 3,000 x g and the resulting pellet was plated onto XLD agar plate (Oxoid, UK) for isolation of bacteria. The plates were then incubated in a 37°C incubator for 24 hours prior to colony identification and counting.