An improved selective and differential medium for the isolation of Burkholderia pseudomallei from clinical specimens

Francis, A.E.; Aiyar, S; Yean, Chan Yean; Naing, Lin; Ravichandran, M.

doi:10.1016/j.diagmicrobio.2005.11.008

Cited by 23 publications

(20 citation statements)

References 11 publications

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“…Aminoglycoside sensitivity was the result of a nonsynonymous SNP within amrB. Most selective media for B. pseudomallei growth contain GEN as a selective agent (26,32,33). Use of these media in Sarawak as a sole diagnostic tool for melioidosis is problematic because, based on our findings, approximately 86% of cases would not be diagnosed.…”

Section: Resultsmentioning

confidence: 68%

Burkholderia pseudomallei Isolates from Sarawak, Malaysian Borneo, Are Predominantly Susceptible to Aminoglycosides and Macrolides

Podin

Sarovich

Price

et al. 2014

Antimicrob Agents Chemother

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h Melioidosis is a potentially fatal disease caused by the saprophytic bacterium Burkholderia pseudomallei. Resistance to gentamicin is generally a hallmark of B. pseudomallei, and gentamicin is a selective agent in media used for diagnosis of melioidosis. In this study, we determined the prevalence and mechanism of gentamicin susceptibility found in B. pseudomallei isolates from Sarawak, Malaysian Borneo. We performed multilocus sequence typing and antibiotic susceptibility testing on 44 B. pseudomallei clinical isolates from melioidosis patients in Sarawak district hospitals. Whole-genome sequencing was used to identify the mechanism of gentamicin susceptibility. A novel allelic-specific PCR was designed to differentiate gentamicin-sensitive isolates from wild-type B. pseudomallei. A reversion assay was performed to confirm the involvement of this mechanism in gentamicin susceptibility. A substantial proportion (86%) of B. pseudomallei clinical isolates in Sarawak, Malaysian Borneo, were found to be susceptible to the aminoglycoside gentamicin, a rare occurrence in other regions where B. pseudomallei is endemic. Gentamicin sensitivity was restricted to genetically related strains belonging to sequence type 881 or its single-locus variant, sequence type 997. Whole-genome sequencing identified a novel nonsynonymous mutation within amrB, encoding an essential component of the AmrAB-OprA multidrug efflux pump. We confirmed the role of this mutation in conferring aminoglycoside and macrolide sensitivity by reversion of this mutation to the wild-type sequence. Our study demonstrates that alternative B. pseudomallei selective media without gentamicin are needed for accurate melioidosis laboratory diagnosis in Sarawak. This finding may also have implications for environmental sampling of other locations to test for B. pseudomallei endemicity.

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Section: Resultsmentioning

confidence: 68%

Burkholderia pseudomallei Isolates from Sarawak, Malaysian Borneo, Are Predominantly Susceptible to Aminoglycosides and Macrolides

Podin

Sarovich

Price

et al. 2014

Antimicrob Agents Chemother

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“…These specimens included blood, pus (wound swabs or ear-nosethroat specimens), tissue, urine and stool, originating from 279 patients who had been hospitalized in various medical and surgical wards for more than 48 h. All blood specimens were isolated using the BD BACTEC TM blood culture system. Each specimen that was identified as Gram negative rods on microscopic examination was directly inoculated onto horseblood agar and MacConkey, (Oxoid, Basingstoke, United Kingdom) and incubated in air at 37°C for 18 to 24 h. Gram negative colonies were inoculated unto Francis media (prepared as in Francis et al, 2006) and homogenized in 1 ml of sterile physiological saline (0.85%) into 0.5 McFarland suspension and plated spirally onto Muller Hinton agar with antibiotic disc (30 µgCefotaxime, 10 µg Amoxicillin-clavulanic acid and 30 µgCeftazidime placed 1.5 cm distance in a row) for ESBL screening and incubated at 35°C for 18 to 24 h. Growth on Francis agar is linked to the resistance to a specific antibiotic and signified by presence of yellow colonies and yellow discoloration of Francis agar were regarded as ESBL isolates (Figure 1).…”

Section: Methodsmentioning

confidence: 99%

“…Francis media was developed for the differential screening of Burkholderia pseudomallei and Burkholderia cepacia (Francis et al, 2006). This media which uses gentamicin as its inhibitory components, was later found to have additional function of selecting ESBL producing isolates.…”

Section: Introductionmentioning

confidence: 99%

Evaluation of Francis media for extended spectrum beta lactamase (ESBL) screening

Ramli¹,

Hashim²,

Francis³

2014

Afr. J. Microbiol. Res.

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Francis media was developed for the differential screening of Burkholderia pseudomallei. It was later found to have additional function of selecting extended spectrum beta lactamase (ESBL) from Enterobacteriaceae. A total of 305 Enterobacteriaceae isolates from clinical specimens (Klebsiella spp. [191], Escherichia coli [96], Enterobacter spp. [9], Citrobacter spp. [3] and others [6]) were tested for the presence of ESBL.Out of 305, 135 were ESBL producing Enterobacteriaceae tested on Francis media for the presence of yellow colonies and haze after 24 h of incubation. Francis media revealed sensitivity and specificity of 89 and 99%, respectively in detecting ESBL producing Enterobactericeae.

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“…Although researchers have extracted and isolated B. pseudomallei from environmental matrices and in clinical settings for decades, the focus has been largely on the selective growth medium rather than extraction procedure (12)(13)(14). However, quantitative extraction and enumeration of microorganisms from soil generally in- corporates a homogenization step where soil is shaken and/or incubated with diluent.…”

Section: Discussionmentioning

confidence: 99%

Survival, Sublethal Injury, and Recovery of Environmental Burkholderia pseudomallei in Soil Subjected to Desiccation

Larsen

Smith

Norton

et al. 2013

Appl Environ Microbiol

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bEnvironmental Burkholderia pseudomallei isolated from sandy soil at Castle Hill, Townsville, in the dry tropic region of Queensland, Australia, was inoculated into sterile-soil laboratory microcosms subjected to variable soil moisture. Survival and sublethal injury of the B. pseudomallei strain were monitored by recovery using culture-based methods. Soil extraction buffer yielded higher recoveries as an extraction agent than sterile distilled water. B. pseudomallei was not recoverable when inoculated into desiccated soil but remained recoverable from moist soil subjected to 91 days' desiccation and showed a growth response to increased soil moisture over at least 113 days. Results indicate that endemic dry tropic soil may act as a reservoir during the dry season, with an increase in cell number and potential for mobilization from soil into water in the wet season.

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An improved selective and differential medium for the isolation of Burkholderia pseudomallei from clinical specimens

Cited by 23 publications

References 11 publications

Burkholderia pseudomallei Isolates from Sarawak, Malaysian Borneo, Are Predominantly Susceptible to Aminoglycosides and Macrolides

Burkholderia pseudomallei Isolates from Sarawak, Malaysian Borneo, Are Predominantly Susceptible to Aminoglycosides and Macrolides

Evaluation of Francis media for extended spectrum beta lactamase (ESBL) screening

Survival, Sublethal Injury, and Recovery of Environmental Burkholderia pseudomallei in Soil Subjected to Desiccation

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