Sr0048

Manufactured by Thermo Fisher Scientific

Sourced in United Kingdom

The SR0048 is a laboratory instrument manufactured by Thermo Fisher Scientific. It is designed for general laboratory use. The core function of the SR0048 is to perform precise measurements and analysis. Detailed specifications and intended use are not available in this response.

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Lab products found in correlation

Sr0147e, by Thermo Fisher Scientific (4 mentions) Biological safety cabinet, by Thermo Fisher Scientific (2 mentions) Cm0331, by Thermo Fisher Scientific (2 mentions) Cm0331b, by Thermo Fisher Scientific (2 mentions) Br0056a, by Thermo Fisher Scientific (2 mentions) P iodonitrotetrazolium chloride int, by Merck Group (1 mentions)

5 protocols using sr0048

Antimicrobial Susceptibility of Clinical Strains

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Doxycycline (200 mg, Combitic Global Caplet Pvt, India), erythromycin (erythromycine stearate 500 mg, Cipla Ltd, India), amoxicillin (amoxicillin trihydrate 500 mg, Maxheal Pharmaceutical Ltd, India) and ciprofloxacin (ZOFLOX, ciprofloxacine 750 mg, Odypharm) used as reference antibiotics were purchased from a local pharmacy. p-Iodonitrotetrazolium chloride (INT, Sigma-Aldrich) was used as a microbial growth indicator (Mativandlela et al. 2006 ). Columbia agar supplemented with 5% (v/v) lacked horse blood (SR0048 Oxoid, Basingstoke, England) and 1% (v/v) Vitox was used for the activation of the tested clinical strains while Brian Heart Infusion (BHI) broth supplemented with 5% horse serum was used for antibacterial assays. CampyGen gas park (CN0024A Oxoid, Basingstoke, England) was used to generate microaerophilic conditions of culture.

Kouitcheu Mabeku L.B., Nanfack Nana B., Eyoum Bille B., Tchuenteu Tchuenguem R, & Nguepi E. (2017). Anti-Helicobacter pylori and antiulcerogenic activity of Aframomum pruinosum seeds on indomethacin-induced gastric ulcer in rats. Pharmaceutical Biology, 55(1), 929-936.

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Isolation and Identification of H. pylori

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The isolation of H. pylori from biopsy specimens was performed as described previously²². Briefly, biopsy specimens were aseptically rolled over the surface of Columbia blood agar base plates under a biological safety cabinet (Thermo Fischer Scientific). The agar (Oxoid CM0331) was supplemented with 7% horse serum (Oxoid SR0048), 1% vitamin mix (Isovitale-X), and an H. pylori selective supplement (Dent, SR0147E Oxoid) comprising of amphotericin B (2.5 mg), trimethoprim (2.5 mg), vancomycin (5.0 mg), and cefsulodin (2.5 mg). The plates were incubated at 37 °C in an atmosphere of 85% N₂, 10% CO₂ and 5% O₂ for 4–10 days. H. pylori colonies were identified as small, round, translucent, Gram-negative bacteria and positive for catalase, oxidase and urease tests. The confirmed isolates were frozen in Brucella broth containing 20% glycerol and stored at −80 °C for future use.

Palamides P., Jolaiya T., Idowu A., Loell E., Onyekwere C., Ugiagbe R., Agbo I., Lesi O., Ndububa D., Adekanle O., Carranza M., Ally R., Njom H., Adeleye I.A., Harrison U., Clarke A., Fischer W., Smith S, & Haas R. (2020). Helicobacter pylori patient isolates from South Africa and Nigeria differ in virulence factor pathogenicity profile and associated gastric disease outcome. Scientific Reports, 10, 11409.

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Isolation and Identification of Helicobacter pylori

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Gastric antral biopsies were placed in 20% glucose solution [23 ], then chopped, and inoculated on Columbia blood agar (CM0331B; Oxoid) supplemented with Dent’s H. pylori selective medium (SR0147E; Oxoid) and 7% laked horse blood (SR0048; Oxoid). Plates were incubated at 37 °C for up to 7 days in a moist microaerophilic atmosphere of 10% CO₂, 5% O₂, and 85% N₂ established using a Campylobacter gas generating kit (BR0056A, Oxoid). After 3 days of initial incubation, plates were evaluated for growth on a daily basis. Where no growth of H. pylori was observed after incubation for 7 days, plates were recorded as negative for H. pylori culture. Following positive growth, individual small rounded, translucent colony-forming units (CFU) were selected and subcultured twice to ensure pure cultures. Isolates were identified as H. pylori on the basis of morphology following Gram stain, positive findings of oxidase, catalase and urease tests as per [24 (link)], and by PCR for 16S rRNA (see Table 1).

Rasheed F., Campbell B.J., Alfizah H., Varro A., Zahra R., Yamaoka Y, & Pritchard D.M. (2014). Analysis of Clinical Isolates of Helicobacter pylori in Pakistan Reveals High Degrees of Pathogenicity and High Frequencies of Antibiotic Resistance. Helicobacter, 19(5), 387-399.

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Culturing H. pylori from Gastric Biopsies

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Biopsy specimens were collected from each patient and dipped into a 20% glucose solution for culturing purposes (12 ). Gastric biopsy specimens were then chopped and inoculated on Columbia blood agar media (CM0331B; Oxoid) supplemented with 7% laked horse blood (SR0048; Oxoid), which also contained Dent’s H. pylori selective medium (SR0147E; Oxoid). The culturing conditions for H. pylori growth were provided by using a Campylobacter gas-generating kit (BR0056A, Oxoid) in which a microaerophilic environment, i.e. 10% carbon dioxide (CO₂), 5% oxygen (O₂), and 85% nitrogen (N₂), was already established. Starting after three days of incubation, the plates were checked on a daily basis for H. pylori growth; if no growth was observed after seven days, the plate was considered negative for H. pylori. Positive cultures were in the form of small rounded colonies, which were sub-cultured to obtain pure cultures. Positive isolates of H. pylori were further identified.

Ahmad S., Ahmad F., Rahman F.U., Khan S., Murad W., Mughal I., ur Rahman A., Muhammad Khan F., Khan I, & Ahmad H. (2016). PCR Based Detection of Phase Variable Genes in Pakistani Based Clinical Helicobacter pylori Strains. Jundishapur Journal of Microbiology, 9(7), e31824.

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Isolation and Culturing of Helicobacter pylori

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Biopsy specimens were aseptically rolled over the surface of Columbia blood agar base plates under a biological safety cabinet (Thermo Scientific). The agar (Oxoid CM0331) was supplemented with 7% horse serum (Oxoid SR0048), 1% vitamin mix (Isovitalex), and an H. pylori selective supplement (Dent, SR0147E Oxoid) comprising of amphotericin B (2.5 mg), trimethoprim (2.5 mg), vancomycin (5.0 mg), and cefsulodin (2.5 mg). The plates were incubated at 37 °C in an atmosphere of 85% N₂, 10% CO₂ and 5% O₂ for 4–10 days. Presumptive H. pylori colonies were identified as small, round, translucent, Gram-negative and positive for catalase, oxidase and urease tests. The confirmed isolates were Frozen in brain heart infusion broth (BHI) containing 20% glycerol and stored at − 80 °C for future use.

Idowu A., Mzukwa A., Harrison U., Palamides P., Haas R., Mbao M., Mamdoo R., Bolon J., Jolaiya T., Smith S., Ally R., Clarke A, & Njom H. (2019). Detection of Helicobacter pylori and its virulence genes (cagA, dupA, and vacA) among patients with gastroduodenal diseases in Chris Hani Baragwanath Academic Hospital, South Africa. BMC Gastroenterology, 19, 73.

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