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Published on: June 5, 2012
The use of selective media for the isolation of Pseudomonas pseudomallei in clinical practice
V Wuthiekanun1, D A Dance, Y Wattanagoon
1Bangkok Hospital for Tropical Diseases, Faculty of Tropical Medicine, Mahidol University, Thailand.
Abstract:
Ashdown's selective-differential agar medium, with or without preenrichment in selective broth, was evaluated for the isolation of Pseudomonas pseudomallei from 1972 clinical specimens obtained from 643 subjects in Northeast Thailand; 226 patients proved to have meliodosis. The use of Ashdown's medium significantly increased the frequency of recovery of P. pseudomallei from sites or specimens with an extensive normal flora (throat, rectum, wounds and sputum) as compared to the recovery on blood and MacConkey agars (p less than 0.01). The isolation frequency from throat, rectal and wound swabs was further increased by the use of the broth pre-enrichment. The colonial morphology of P. pseudomallei on Ashdown's medium was sufficiently characteristic to allow presumptive identification. With the use of these selective media it was possible to culture P. pseudomallei from throat swabs taken from 87% of the patients from whom the organism could also be isolated from corresponding tracheal aspirates or sputum specimens. P. pseudomallei was isolated from rectal swabs taken from 51 patients, the first time that faecal excretion of the organism has been demonstrated in man. The diagnosis of melioidosis would not have been confirmed bacteriologically in eight patients (3.5%) without the use of the selective media. It is suggested that, in areas endemic for melioidosis, all sputum specimens should be cultured on selective media, such as Ashdown's. For the investigation of clinically suspected cases of melioidosis, and for follow-up during treatment of the disease, the use of broth pre-enrichment is recommended for specimens obtained from sites with an extensive normal flora.
Insights
Ashdown's selective medium enhances isolation of Pseudomonas pseudomallei from clinical samples, especially those with normal flora. Broth pre-enrichment further improves recovery rates for melioidosis diagnosis.
Area of Science:
- Medical Microbiology
- Infectious Diseases
- Clinical Diagnostics
Background:
- Melioidosis, caused by Pseudomonas pseudomallei, is endemic in Northeast Thailand.
- Accurate and timely diagnosis is crucial for effective treatment and patient outcomes.
- Traditional culture methods may have limitations in isolating P. pseudomallei from diverse clinical specimens.
Purpose of the Study:
- To evaluate Ashdown's selective-differential agar medium for isolating Pseudomonas pseudomallei.
- To assess the impact of broth pre-enrichment on isolation frequency.
- To compare the efficacy of Ashdown's medium with standard blood and MacConkey agars.
Main Methods:
- Evaluation of Ashdown's medium with and without broth pre-enrichment.
- Culture of 1972 clinical specimens from 643 subjects in Northeast Thailand.
- Comparison of isolation rates on Ashdown's medium versus blood and MacConkey agars.
Main Results:
- Ashdown's medium significantly increased P. pseudomallei recovery from sites with normal flora (p < 0.01).
- Broth pre-enrichment further improved isolation from throat, rectal, and wound swabs.
- Characteristic colonial morphology on Ashdown's medium allowed presumptive identification.
- P. pseudomallei was isolated from rectal swabs, indicating fecal excretion in humans.
- Selective media were essential for confirming melioidosis in 3.5% of patients.
Conclusions:
- Ashdown's selective medium is highly effective for isolating Pseudomonas pseudomallei, particularly from specimens with mixed flora.
- Broth pre-enrichment is recommended for optimizing isolation from specific sites.
- The use of selective media, like Ashdown's, is crucial for accurate melioidosis diagnosis in endemic areas.
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