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  • An improved selective culture medium enhances the isolation of Burkholderia pseudomallei from contaminated specimens.

An improved selective culture medium enhances the isolation of Burkholderia pseudomallei from contaminated specimens.

The American journal of tropical medicine and hygiene (2013-09-26)
Andrew Goodyear, Linda Strange, Drew A Rholl, Joy Silisouk, David A B Dance, Herbert P Schweizer, Steven Dow
ABSTRACT

Burkholderia pseudomallei is a Gram-negative environmental bacterium found in tropical climates that causes melioidosis. Culture remains the diagnostic gold standard, but isolation of B. pseudomallei from heavily contaminated sites, such as fecal specimens, can be difficult. We recently reported that B. pseudomallei is capable of infecting the gastrointestinal tract of mice and suggested that the same may be true in humans. Thus, there is a strong need for new culture techniques to allow for efficient detection of B. pseudomallei in fecal and other specimens. We found that the addition of norfloxacin, ampicillin, and polymyxin B to Ashdown's medium (NAP-A) resulted in increased specificity without affecting the growth of 25 B. pseudomallei strains. Furthermore, recovery of B. pseudomallei from human clinical specimens was not affected by the three additional antibiotics. Therefore, we conclude that NAP-A medium provides a new tool for more sensitive isolation of B. pseudomallei from heavily contaminated sites.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Ampicillin trihydrate, 900-1050 μg/mg anhydrous basis (HPLC)
Sigma-Aldrich
Polymyxin B sulfate salt
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Polymyxin B sulfate salt, powder, BioReagent, suitable for cell culture
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Ampicillin, meets USP testing specifications
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Polymyxin B solution, 20 mg/mL in H2O
Supelco
Polymyxin B solution, 1 mg/mL in H2O, analytical standard
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Ampicillin trihydrate, VETRANAL®, analytical standard
Sigma-Aldrich
Ampicillin sodium salt, BioXtra, suitable for cell culture
Sigma-Aldrich
Ampicillin sodium salt, powder or crystals, BioReagent, suitable for cell culture
Sigma-Aldrich
Polymyxin B sulfate, meets USP testing specifications
Supelco
Norfloxacin, analytical standard, ≥98% (TLC)
Sigma-Aldrich
Ampicillin, anhydrous, 96.0-102.0% (anhydrous basis)
Sigma-Aldrich
Ampicillin sodium salt
Polymyxin B sulfate, European Pharmacopoeia (EP) Reference Standard
Supelco
Norfloxacin, VETRANAL®, analytical standard
Supelco
Ampicillin, analytical standard