Potential of recombinant flagellin fragment from Burkholderia thailandensis as an antigen for melioidosis antibody detection by indirect ELISA

Wajanarogana, Sumet; Nimnuch, Phitchapat; Thongmee, Acharawan; Kritsiriwuthinan, Kanyanan

doi:10.1016/j.mcp.2012.11.001

Cited by 9 publications

(12 citation statements)

References 31 publications

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“…For example, flagellin has been used for a generation of vaccines against Salmonella enterica serovar Paratyphi A, P. aeruginosa , and E. coli O157:H7 [96,97,98], and the colonization of EHEC in cattle was significantly decreased after vaccination with FliC [97]. Due to its high immunogenicity, flagellin has also been used as a vaccine adjuvant together with poorly immunogenic antigens, and FliC or anti-FliC antibodies have been used in diagnostics of e.g., melioidosis and inflammatory bowel disease [99]. Interestingly, flagellin has also been shown to suppress apoptosis and protect against radiation-related damage [100].…”

Section: Discussionmentioning

confidence: 99%

The Role of the Bacterial Flagellum in Adhesion and Virulence

Haiko

Westerlund-Wikström

2013

Biology

454

345

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The bacterial flagellum is a complex apparatus assembled of more than 20 different proteins. The flagellar basal body traverses the cell wall, whereas the curved hook connects the basal body to the whip-like flagellar filament that protrudes several µm from the bacterial cell. The flagellum has traditionally been regarded only as a motility organelle, but more recently it has become evident that flagella have a number of other biological functions. The major subunit, flagellin or FliC, of the flagellum plays a well-documented role in innate immunity and as a dominant antigen of the adaptive immune response. Importantly, flagella have also been reported to function as adhesins. Whole flagella have been indicated as significant in bacterial adhesion to and invasion into host cells. In various pathogens, e.g., Escherichia coli, Pseudomonas aeruginosa and Clostridium difficile, flagellin and/or the distally located flagellar cap protein have been reported to function as adhesins. Recently, FliC of Shiga-toxigenic E. coli was shown to be involved in cellular invasion via lipid rafts. Here, we examine the latest or most important findings regarding flagellar adhesive and invasive properties, especially focusing on the flagellum as a potential virulence factor.

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Section: Discussionmentioning

confidence: 99%

The Role of the Bacterial Flagellum in Adhesion and Virulence

Haiko

Westerlund-Wikström

2013

Biology

454

345

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“…Both of the organisms are broadly similar genetically and immunologically (Yu et al 2006 ). Previous studies indicate that melioidosis protective antibodies can be raised using antigens from B. thailandensis (Ngugi et al 2010 ; Scott et al 2013 ) and certain antigens of B. pseudomallei share epitopes with this closely related species (Wajanarogana et al 2013 ). In addition, some biological properties of B. pseudomallei have been explained by using B. thailandensis as a non-pathogenic model (Haraga et al 2008 ).…”

Section: Introductionmentioning

confidence: 99%

“…A number of serodiagnostic antigens of B. pseudomallei have been revealed using protein microarray (Felgner et al 2009 ) and indirect enzyme-linked immunosorbent assays (ELISA) (Allwood et al 2008 ; Arora et al 2015 ; Wajanarogana et al 2013 ; Hara et al 2013 ). The majority of such antigens are cell surface-associated proteins such as outer membrane proteins (Omps), and secreted molecules.…”

Section: Introductionmentioning

confidence: 99%

“…In this study, we focused on Omps, especially outer membrane protein A (OmpA) and truncated flagellin fragment (FLAG300) of B. thailandensis E264. The FLAG300 indicates as the potential antigen for diagnosis of melioidosis (Wajanarogana et al 2013 ); however, culture-confirmed melioidosis serum samples have not yet been studied. As well as a flagellin fragment, OmpA of B. pseudomallei has immunogenic properties which has been used for serodiagnosis of melioidosis (Allwood et al 2008 ; Arora et al 2015 ; Hara et al 2013 ).…”

Section: Introductionmentioning

confidence: 99%

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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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“…The OmpA-based indirect ELISA exhibited a sensitivity of 82.6-95% and a specificity of 93.75-98% [1,2]. In addition, Chen et al and Wajanarogana et al demonstrated that indirect ELISA applying FliC as the antigen achieved 82.7-93.8% sensitivity and 94.6-96.3% specificity and offered a more efficient serodiagnosis of melioidosis [8,25]. However, the differences in specificity and sensitivity between the two proposed probes in goat melioidosis remain unknown.…”

mentioning

confidence: 99%

Development of an immunoassay using recombinant outer membrane protein A and flagellin for diagnosis of goats with melioidosis

Lee

Shih

et al. 2020

The Journal of Veterinary Medical Science

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Among domestic animals, melioidosis is one of the most common diseases reported in goat, sheep, and swine. To evaluate the specific antibodies in goats with melioidosis, we developed a serology test using recombinant outer membrane protein A (OmpA) and flagellin (FliC) of Burkholderia pseudomallei as antigens. DNA corresponding to each antigen was cloned into a pET32a vector and expressed in Escherichia coli. Essentially, the recombinant OmpA and FliC were expressed in a soluble form that could be isolated with 95% homogeneity. Both recombinants could be recognized by rabbit antibodies prepared against heat-inactivated B.

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Potential of recombinant flagellin fragment from Burkholderia thailandensis as an antigen for melioidosis antibody detection by indirect ELISA

Cited by 9 publications

References 31 publications

The Role of the Bacterial Flagellum in Adhesion and Virulence

The Role of the Bacterial Flagellum in Adhesion and Virulence

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

Development of an immunoassay using recombinant outer membrane protein A and flagellin for diagnosis of goats with melioidosis

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