Improved diagnosis of melioidosis using a 2-dimensional immunoarray

Sorenson, Alanna E.; Williams, Natasha L.; Morris, Jodie L.; Ketheesan, Natkunam; Norton, Robert; Schaeffer, Patrick M.

doi:10.1016/j.diagmicrobio.2013.07.009

Cited by 9 publications

(10 citation statements)

References 27 publications

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“…Sorenson et al developed and evaluated a two-dimensional immunoarray (2DIA) designed to detect the three most common Burkholderia LPS types (A, B, and B2). Using a panel of 60 serum samples, they found that 2DIA had a much higher sensitivity than IHA (100 vs. 58.6 %), giving positive results for all melioidosis culture-positive/IHA-negative serums, but a lower specificity (87.1 vs. 100 %) [45]. …”

Section: Diagnosticsmentioning

confidence: 99%

Recent Advances in Burkholderia mallei and B. pseudomallei Research

2015

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Burkholderia mallei and Burkholderia pseudomallei are Gram-negative organisms, which are etiological agents of glanders and melioidosis, respectively. Although only B. pseudomallei is responsible for a significant number of human cases, both organisms are classified as Tier 1 Select Agents and their diseases lack effective diagnosis and treatment. Despite a recent resurgence in research pertaining to these organisms, there are still a number of knowledge gaps. This article summarizes the latest research progress in the fields of B. mallei and B. pseudomallei pathogenesis, vaccines, and diagnostics.

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Section: Diagnosticsmentioning

confidence: 99%

Recent Advances in Burkholderia mallei and B. pseudomallei Research

2015

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“…Previous studies reported that B. pseudomallei strains producing different LPS types are epidemiologically different. 20 , 21 The majority of B. pseudomallei strains are of the typical LPS type; however, 14.7% and 2.3% of strains isolated from northern Australia and southeast Asia, respectively, are of the atypical type (B, B2, or rough type). 19 Distribution of B. pseudomallei strains in newly identified endemic areas such as the Indian subcontinent, southern China, Hong Kong, and Taiwan is still largely unknown.…”

Section: Introductionmentioning

confidence: 99%

Development of Immunoassays for Burkholderia pseudomallei Typical and Atypical Lipopolysaccharide Strain Typing

Nualnoi

Norris

Tuanyok

et al. 2017

The American Society of Tropical Medicine and Hygiene

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Abstract. Burkholderia pseudomallei is the causative agent of melioidosis, a severe infection endemic to many tropical regions. Lipopolysaccharide (LPS) is recognized as an important virulence factor used by B. pseudomallei. Isolates of B. pseudomallei have been shown to express one of four different types of LPS (typical LPS, atypical LPS types B and B2, and rough LPS) and in vitro studies have demonstrated that LPS types may impact disease severity. The association between LPS types and clinical manifestations, however, is still unknown, in part because an effective method for LPS type identification is not available. Thus, we developed antigen capture immunoassays capable of distinguishing between the LPS types. Mice were injected with B or B2 LPS for atypical LPS-specific monoclonal antibody (mAb) isolation; only two mAbs (3A2 and 5B4) were isolated from mice immunized with B2 LPS. Immunoblot analysis and surface plasmon resonance demonstrated that 3A2 and 5B4 are reactive with both B2 and B LPS where 3A2 was shown to possess higher affinity. Assays were then developed using capsular polysaccharidespecific mAb 4C4 for bacterial capture and 4C7 (previously shown to bind typical LPS) or 3A2 mAbs for typical or atypical LPS strain detection, respectively. The evaluations performed with 197 strains of Burkholderia and nonBurkholderia species showed that the assays are reactive to B. pseudomallei and Burkholderia mallei strains and have an accuracy of 98.8% (zero false positives and two false negatives) for LPS typing. The results suggest that the assays are effective and applicable for B. pseudomallei LPS typing.

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“…Moreover, bacterial culture is an imperfect reference method because it has low sensitivity and negative predictive value (Limmathurotsakul et al 2010 ). Serological diagnosis of melioidosis is widely used in endemic regions, especially the indirect hemagglutination assay (IHA); however, it has limited utility for diagnosis because of relatively poor sensitivity (Appassakij et al 1990 ; Harris et al 2009 ; Sorenson et al 2013 ) and a high seropositivity background in endemic areas (Ashdown and Guard 1984 ; Kanaphun et al 1993 ). Although it is of limited value for diagnosis in those areas, it may be of assistance in some circumstances, particularly when paired sera are available or high serum titers are reported with the presence of clinical signs.…”

Section: Introductionmentioning

confidence: 99%

“…Although it is of limited value for diagnosis in those areas, it may be of assistance in some circumstances, particularly when paired sera are available or high serum titers are reported with the presence of clinical signs. Currently, a number of serodiagnostic tests for melioidosis detection has been developed using crude extracts (Sorenson et al 2013 ; Cooper et al 2013 ; Parthasarathy et al 2006 ) or purified recombinant proteins from B. pseudomallei (Allwood et al 2008 ; Arora et al 2015 ; Chantratita et al 2007 ; Chen et al 2003 ; Druar et al 2008 ) as antigens. Even though they obviously demonstrate substantial improvement over the clinical standard IHA test, all antigens are derived from a Tier 1 agent, B. pseudomallei , and thus manipulations must be done under biosafety level 3 (BSL-3).…”

Section: Introductionmentioning

confidence: 99%

Efficacy of indirect ELISA for serodiagnosis of melioidosis using immunodominant antigens from non-pathogenic Burkholderia thailandensis

Wajanarogana

Kritsiriwuthinan

2016

SpringerPlus

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Melioidosis caused by gram negative bacteria, B. pseudomallei, is a fatal disease in the tropical and sub-tropical regions. However, sporadic cases have been reported in elsewhere. Early detection is imperative to reduce the mortality rate. Serological tests have being substantially developed using recombinant proteins as specific targeted antigens to melioidosis antibodies. In the present study, we focus on a truncated flagellin fragment (FLAG300) and outer membrane protein A (OmpABT) of B. thailandensis E264 as potential antigens for developing indirect ELISA to improve the serodiagnosis of melioidosis. Recombinant proteins were overexpressed and purified by immobilized metal affinity chromatography with denaturing conditions. The sensitivity and specificity of individual test were calculated within culture-confirmed melioidosis sera (n = 42) and non-melioidosis serum samples (n = 241) using the cut-off point at average of absorbance plus 2 standard deviations. The results demonstrated that a FLAG 300 based indirect ELISA showed 90.48 % sensitivity and 87.14 % specificity and an OmpABT based this assay revealed sensitivity of 80.95 % and specificity of 89.21 %. Their use in a double-antigen ELISA resulted in improve specificity (92.95 %) and still high degree of sensitivity (85.71 %). These data suggest a facile method for serodiagnosis of melioidosis by the use of antigens from a non-pathogenic strain.

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Improved diagnosis of melioidosis using a 2-dimensional immunoarray

Cited by 9 publications

References 27 publications

Recent Advances in Burkholderia mallei and B. pseudomallei Research

Recent Advances in Burkholderia mallei and B. pseudomallei Research

Development of Immunoassays for Burkholderia pseudomallei Typical and Atypical Lipopolysaccharide Strain Typing

Efficacy of indirect ELISA for serodiagnosis of melioidosis using immunodominant antigens from non-pathogenic Burkholderia thailandensis

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