Evaluation of PCR, DNA hybridization and immunomagnetic separation — PCR for detection of Burkholderia mallei in artificially inoculated environmental samples

Systematic monitoring of glanders-infected horses by complement fixation test, bacterial isolation, and PCR

“…A suspension of each clinical sample for microbiological analysis was seeded on 5% sheep blood agar with 5% glycerin and 2500 IU potassium benzylpenicillin (selective medium for B. mallei ) and incubated for 48–72 hours at 37°C ( Quinn et al., 2005 ; Winn et al., 2008 ; Merwyn et al., 2010 ). Morphological characteristics of the resultant bacterial colonies were recorded, including size, shape, color, and presence and type of hemolysis.…”

“…Isolating B. mallei from clinical specimens, mainly nasal swabs, presents a challenge due to high occurrence of other bacteria ( Merwyn, Kumar, Agarwal and Rai, 2010 ). This is compounded by the low bacterial load usually present in infected equines, which generally do not show clinical manifestation of chronic disease ( Scholz et al., 2006 ).…”

“…Polymerase chain reaction (PCR) can be an important tool for detecting B. mallei , since it has high sensitivity and does not depend on the viability of the agent for its identification ( Merwyn et al., 2010 ). Detection of B. mallei by molecular methods is recommended for both acute and chronic manifestations, due to the rapid identification of the agent, differential diagnosis from melioidosis Burkholderia pseudomallei , and conclusiveness of the diagnosis ( Suppiah, Thimma, Cheah and Vadivelu, 2009 ; OIE, 2015 ).…”

Burkholderia Mallei Isolation From Milk of a Mare and Evidence of Congenital Transmission of Glanders in Equids: Case Reports

“…The bagged organ and milk samples were suspended 1:5 w/vin sterile 0.85% saline solution, and 10 µL of the suspension was seeded onto 5% sheep blood agar with 5% glycerin and 2500 IU potassium benzyl penicillin (selective medium for B. mallei) and incubated for 48-72 h at 37°C (MERWYN et al, 2010;QUINN et al, 2011;WINN et al, 2008). Morphological characteristics of the resultant bacterial colonies were recorded, including size, shape, color, and presence and type of hemolysis.…”

“…A total of 23 samples were cultured for bacteria isolation: nine from horse #1, six from the fetus of horse #1, and eight from horse #2 (Table 1). Only the milk sample from horse #2 showed growth of B. mallei in a blood enriched medium (selective), suggesting that using this protocol could be more appropriate to isolation of B. mallei compared with other conventional isolation methods (MERWYN et al, 2010). Growth of Bacillus spp., Staphylococcus spp., Escherichia coli, Pantoea agglomerans, and yeasts (contaminants) was observed, with eight samples showing no growth after bacteriological culture.…”