Publication: Rapid identification of Burkholderia pseudomallei in blood cultures by a monoclonal antibody assay
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Issued Date
1999
Resource Type
File Type
application/pdf
ISSN
951137
Other identifier(s)
2-s2.0-0032834666
Rights Holder(s)
มหาวิทยาลัยศรีนครินทรวิโรฒ
Bibliographic Citation
Journal of Clinical Microbiology. Vol 37, No.11 (1999), p.3662-3667
Suggested Citation
Pongsunk S., Thirawattanasuk N., Piyasangthong N., Ekpo P. Rapid identification of Burkholderia pseudomallei in blood cultures by a monoclonal antibody assay. Journal of Clinical Microbiology. Vol 37, No.11 (1999), p.3662-3667. doi:10.1128/jcm.37.11.3662-3667.1999 Retrieved from: https://hdl.handle.net/20.500.14740/7178
Author(s)
Abstract
Burkholderia pseudomallei is the causative agent of melioidosis. In northeast Thailand, this gram-negative bacterium is a major cause of mortality from septicemia. The definitive diagnosis of this disease is made by bacterial culture. In this study, we produced a monoclonal antibody (MAb) specific to the 30-kDa protein of B. pseudomallei by in vivo and in vitro immunization of BALB/c mice with a crude culture filtrate antigen. The MAb could directly agglutinate with all 243 clinical isolates of B. pseudomallei but not with other gram-negative bacteria, except for one strain of Burkholderia mallei. However, the MAb cross-reacted with the gram-positive Bacillus sp. and Streptococcus pyogenes. B. pseudomallei in brain heart infusion broth (BHIB) subcultured from a BacT/Alert automated blood culture system could be identified by simple agglutination with this MAb assay. The sensitivity and specificity of direct agglutination compared to the 'gold standard,' the culture method, were 94.12 and 98.25%, respectively. However, the MAb adsorbed to polystyrene beads or latex particles directly identified the bacterium in blood culture specimens and in BHIB subcultured from a BacT/Alert automated blood culture system. The sensitivity of the latex agglutination test was 100% for both blood culture and BHIB specimens. The specificity was 85.96 and 96.49% for the blood culture and BHIB specimens, respectively. The specificity could be increased if the nonspecific materials in the blood culture broths were eradicated by centrifugation at low speeds. Thus, a combination of blood culture and the agglutination method could be used for the rapid diagnosis of melioidosis in the routine bacteriological laboratory. This method could speed up detection of the bacterium in blood culture by at least 2 days, compared to the conventional bacterial culture method. In addition, the MAb is stable at room temperature for 2 weeks and at 4, -20, and -70°C for at least 1 year. The latex reagent was stable for at least 6 months at 4°C.
Subject(s)
Monoclonal antibody
Antibiotic resistance
Article
Bacillus
Bacterium culture
Bacterium identification
Blood culture
Burkholderia pseudomallei
DNA determination
Enzyme linked immunosorbent assay
Gram negative bacterium
Immunization
Immunoblotting
Melioidosis
Mortality
Priority journal
Septicemia
Streptococcus pyogenes
Thailand
Burkholderia pseudomallei
Antibiotic resistance
Article
Bacillus
Bacterium culture
Bacterium identification
Blood culture
Burkholderia pseudomallei
DNA determination
Enzyme linked immunosorbent assay
Gram negative bacterium
Immunization
Immunoblotting
Melioidosis
Mortality
Priority journal
Septicemia
Streptococcus pyogenes
Thailand
Burkholderia pseudomallei