Vibrio in the specimens was enriched with alkaline peptone water (APW; Luqiao, Beijing, China) and selected on thiosulfate-citrate-bile-sucrose (TCBS) agar (Oxoid, Basingstoke, UK). Salmonella was enriched with selenite brilliant green (SBG) broth (Luqiao) and selected on Salmonella Chromogenic Medium (Kemajia, Shanghai, China). Suspicious colonies were identified using serum agglutination tests and biochemical reactions with VITEK 2 Gram-Negative Identification (GN) cards (bioMérieux, France). One milliliter of an overnight culture of APW and SBG was stored at –80°C for nucleic acid extraction.
Vitek 2 gram negative identification gn cards
Manufactured by bioMérieux
Sourced in France
The VITEK 2 Gram-Negative Identification (GN) cards are a laboratory product designed for the identification of gram-negative bacteria. The cards provide a standardized and automated method for the rapid identification of a wide range of gram-negative microorganisms.
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Lab products found in correlation
2 protocols using vitek 2 gram negative identification gn cards
1
Vibrio and Salmonella Isolation Protocol
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Vibrio in the specimens was enriched with alkaline peptone water (APW; Luqiao, Beijing, China) and selected on thiosulfate-citrate-bile-sucrose (TCBS) agar (Oxoid, Basingstoke, UK). Salmonella was enriched with selenite brilliant green (SBG) broth (Luqiao) and selected on Salmonella Chromogenic Medium (Kemajia, Shanghai, China). Suspicious colonies were identified using serum agglutination tests and biochemical reactions with VITEK 2 Gram-Negative Identification (GN) cards (bioMérieux, France). One milliliter of an overnight culture of APW and SBG was stored at –80°C for nucleic acid extraction.
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Vibrio in the specimens was enriched with alkaline peptone water (APW; Luqiao, Beijing, China) and selected on thiosulfate-citrate-bile-sucrose (TCBS) agar (Oxoid, Basingstoke, UK). Salmonella was enriched with selenite brilliant green (SBG) broth (Luqiao) and selected on Salmonella Chromogenic Medium (Kemajia, Shanghai, China). Suspicious colonies were identified using serum agglutination tests and biochemical reactions with VITEK 2 Gram-Negative Identification (GN) cards (bioMérieux, France). One milliliter of an overnight culture of APW and SBG was stored at –80°C for nucleic acid extraction.
2
Comprehensive Enteric Pathogen Screening
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The following pathogens were tested for in every sample by strain isolation: Salmonella, Shigella, diarrheogenic Escherichia coli (DEC), pathogenic Vibrio, Aeromonas and Plesiomonas shigelloides.
Strains of Salmonella were cultured in Selenite Brilliant Green Broth (SBG) and selected on Salmonella Chromogenic Medium. Strains of Vibrio, Aeromonas, and P. shigelloides were cultured in Alkaline Peptone Water (APW) and selected on thiosulfate-citratebile-sucrose (TCBS) agar and MacKenzie (Mac). The fecal samples were streaked on Mac and Xylose Lysine Deoxycholate medium (XLD) for isolation of DEC and Shigella. The suspected colonies of Salmonella and Shigella were examined using serum agglutination tests.
For the identification of DEC strains, 10 suspected E. coli colonies on Mac and XLD were picked and mixed in 200 ml of deioned water.
The mixture was screened by real-time PCR (Hidaka et al., 2009) for pathogenic genes eae, stx1, elt, sth, stp, aggR, virB, and ipaH. Once positive results occurred, the colonies were detected separately to identify the DEC strains.
All suspected colonies were identified by systematic biochemical reaction with VITEK 2 Gram-Negative Identification (GN) cards (BioMerieux, France). All media in the study were obtained from OXOID, UK.
Strains of Salmonella were cultured in Selenite Brilliant Green Broth (SBG) and selected on Salmonella Chromogenic Medium. Strains of Vibrio, Aeromonas, and P. shigelloides were cultured in Alkaline Peptone Water (APW) and selected on thiosulfate-citratebile-sucrose (TCBS) agar and MacKenzie (Mac). The fecal samples were streaked on Mac and Xylose Lysine Deoxycholate medium (XLD) for isolation of DEC and Shigella. The suspected colonies of Salmonella and Shigella were examined using serum agglutination tests.
For the identification of DEC strains, 10 suspected E. coli colonies on Mac and XLD were picked and mixed in 200 ml of deioned water.
The mixture was screened by real-time PCR (Hidaka et al., 2009) for pathogenic genes eae, stx1, elt, sth, stp, aggR, virB, and ipaH. Once positive results occurred, the colonies were detected separately to identify the DEC strains.
All suspected colonies were identified by systematic biochemical reaction with VITEK 2 Gram-Negative Identification (GN) cards (BioMerieux, France). All media in the study were obtained from OXOID, UK.
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