Rapid Detection of Burkholderia pseudomallei in Blood Cultures Using a Monoclonal Antibody-Based Immunofluorescent Assay

Chantratita, Narisara; Tandhavanant, Sarunporn; Wongsuvan, Gumphol; Wuthiekanun, Vanaporn; Teerawattanasook, Nittaya; Day, Nicholas P. J.; Limmathurotsakul, Direk; Peacock, Sharon J.

doi:10.4269/ajtmh.13-0212

Cited by 24 publications

(23 citation statements)

References 9 publications

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“…These characteristics are comparable to the original report of this test elsewhere in Thailand, 5 confirming validity of the assay. Notably, in the previous study the IFA was performed on all positive blood cultures, whereas in this study we restricted testing to positive blood cultures with Gram-negative organisms.…”

Section: Discussionsupporting

confidence: 86%

Validation of a monoclonal antibody-based immunofluorescent assay to detect Burkholderia pseudomallei in blood cultures

Dulsuk

Paksanont

Sangchankoom

et al. 2017

Trans R Soc Trop Med Hyg

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BackgroundIdentification of Burkholderia pseudomallei, the cause of melioidosis, using routine methods takes several days. Use of a monoclonal antibody-based immunofluorescent assay (IFA) on positive blood cultures may speed diagnosis.MethodsWe tested the diagnostic accuracy of the IFA on 545 blood cultures positive for Gram-negative organisms at Udon Thani Hospital, Thailand, between June 2015 and August 2016.ResultsSensitivity of the IFA was 100% and specificity was 99.6%. The median decrease in time to pathogen identification between the IFA result and routine methods was 28 h (IQR 25–51), p<0.0001.ConclusionsThe IFA accurately expedites the diagnosis of melioidosis.

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Section: Discussionsupporting

confidence: 86%

Validation of a monoclonal antibody-based immunofluorescent assay to detect Burkholderia pseudomallei in blood cultures

Dulsuk

Paksanont

Sangchankoom

et al. 2017

Trans R Soc Trop Med Hyg

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“…pseudomallei CPS were developed. These diagnostic tools showed similar specificity but higher sensitivity when compared to the ‘gold standard’ diagnostic bacterial culture, suggesting that these techniques have the potential to be used clinically [7,10,23]. To use these tools as a routine diagnostic method, however, it is important to understand the fate of CPS in vivo .…”

Section: Discussionmentioning

confidence: 99%

In vivo Distribution and Clearance of Purified Capsular Polysaccharide from Burkholderia pseudomallei in a Murine Model

et al. 2016

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Burkholderia pseudomallei is the causative agent of melioidosis, a severe infection prominent in northern Australia and Southeast Asia. The “gold standard” for melioidosis diagnosis is bacterial isolation, which takes several days to complete. The resulting delay in diagnosis leads to delayed treatments, which could result in death. In an attempt to develop better methods for early diagnosis of melioidosis, B. pseudomallei capsular polysaccharide (CPS) was identified as an important diagnostic biomarker. A rapid lateral flow immunoassay utilizing CPS-specific monoclonal antibody was developed and tested in endemic regions worldwide. However, the in vivo fate and clearance of CPS has never been thoroughly investigated. Here, we injected mice with purified CPS intravenously and determined CPS concentrations in serum, urine, and major organs at various intervals. The results indicate that CPS is predominantly eliminated through urine and no CPS accumulation occurs in the major organs. Immunoblot analysis demonstrated that intact CPS was excreted through urine. To understand how a large molecule like CPS was eliminated without degradation, a 3-dimenational structure of CPS was modeled. The predicted CPS structure has a rod-like shape with a small diameter that could allow it to flow through the glomerulus of the kidney. CPS clearance was determined using exponential decay models and the corrected Akaike Information Criterion. The results show that CPS has a relatively short serum half-life of 2.9 to 4.4 hours. Therefore, the presence of CPS in the serum and/or urine suggests active melioidosis infection and provides a marker to monitor treatment of melioidosis.

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“…120 Alternatively, flagged blood cultures or bacterial colonies on culture plates can now rapidly and accurately be identified using real-time PCR targeting the B. pseudomallei type III secretion system (TTS1) gene cluster 121 or using various locally developed antigen detection systems that are not widely available, such as B. pseudomallei-specific latex agglutination and immunofluorescence. 122,123 Direct real-time PCR assays of clinical samples have been trialed but, while providing a more rapid diagnosis, they have to date been less sensitive than blood cultures for detecting bacteremic melioidosis. [124][125][126] Rapid immunofluorescence microscopy of pus, sputum, and urine has been useful in Thailand for rapid diagnosis but is not generally available elsewhere.…”

Section: Diagnosis: Culture Of B Pseudomallei Remains the Gold Standardmentioning

confidence: 99%

Melioidosis: Evolving Concepts in Epidemiology, Pathogenesis, and Treatment

Currie

2015

Semin Respir Crit Care Med

275

347

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Infection with Burkholderia pseudomallei can result in asymptomatic seroconversion, a single skin lesion that may or may not heal spontaneously, a pneumonia which can be subacute or chronic and mimic tuberculosis or rapidly progressive resulting in fatal overwhelming sepsis. Latency with subsequent activation of disease is well recognized, but very uncommon. Melioidosis also has a myriad of other clinical presentations and diagnosis is often delayed because of this and because of difficulties with laboratory diagnosis and lack of recognition outside melioidosis-endemic regions. The perception of B. pseudomallei as a top tier biothreat agent has driven large funding for research, yet resources for diagnosis and therapy of melioidosis in many endemic locations remain extremely limited, with mortality as high as 50% in comparison to around 10% in regions where state-of-the-art intensive care therapy for sepsis is available. Fatal melioidosis is extremely unlikely from natural infection in a healthy person, provided the diagnosis is made early, ceftazidime or meropenem is commenced and intensive care therapy is available. While biothreat research is directed toward potential aerosol exposure to B. pseudomallei, the overall proportion of melioidosis cases resulting from inhalation rather than from percutaneous inoculation remains entirely uncertain, although the epidemiology supports a shift to inhalation during severe weather events such as cyclones and typhoons. What makes B. pseudomallei such a dangerous organism for patients with diabetes and other selective risk factors remains unclear, but microbial genome-wide association studies linking clinical aspects of melioidosis cases to nonubiquitous or polymorphic B. pseudomallei genes or genomic islands are beginning to uncover specific virulence signatures. Finally, what also remains uncertain is the global phylogeography of B. pseudomallei and whether melioidosis is spreading beyond historical locations or is just being unmasked in Africa and the Americas by better recognition and increased surveillance.

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Rapid Detection of Burkholderia pseudomallei in Blood Cultures Using a Monoclonal Antibody-Based Immunofluorescent Assay

Cited by 24 publications

References 9 publications

Validation of a monoclonal antibody-based immunofluorescent assay to detect Burkholderia pseudomallei in blood cultures

Validation of a monoclonal antibody-based immunofluorescent assay to detect Burkholderia pseudomallei in blood cultures

In vivo Distribution and Clearance of Purified Capsular Polysaccharide from Burkholderia pseudomallei in a Murine Model

Melioidosis: Evolving Concepts in Epidemiology, Pathogenesis, and Treatment

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