Development of an enzyme-linked immunosorbent assay for detecting <i>Campylobacter hepaticus</i> specific antibodies in chicken sera – a key tool in Spotty Liver Disease screening and vaccine development

Muralidharan, Chithralekha; Anwar, Arif; Wilson, Timothy B.; Scott, Peter C.; Moore, Robert J.; Van, Thi Thu Hao

doi:10.1080/03079457.2020.1813252

Cited by 8 publications

(12 citation statements)

References 22 publications

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“…The recombinant protein fragment used in SLD-ELISA2 specifically detect anti- C. hepaticus antibodies in bird sera and was not cross-reactive with anti- Campylobacter jejuni or Campylobacter coli antibodies. SLD-ELISA1 showed 96% specificity and 98% sensitivity ( 23 ) whereas SLD-ELISA2 showed 95% specificity and 93% sensitivity (to be published elsewhere). Cross-reactivity of SLD-ELISA2 with C. bilis anti-FHA antibody is yet to be determined.…”

Section: Introductionmentioning

confidence: 96%

“…Two ELISAs have been developed to detect C. hepaticus specific antibodies, namely SLD-ELISA1, based on C. hepaticus total proteins ( 23 ) and SLD-ELISA2, based on a recombinant immunogenic fragment of filamentous hemagglutinin adhesin (FHA) protein of C. hepaticus (paper submitted for publication). The recombinant protein fragment used in SLD-ELISA2 specifically detect anti- C. hepaticus antibodies in bird sera and was not cross-reactive with anti- Campylobacter jejuni or Campylobacter coli antibodies.…”

Section: Introductionmentioning

confidence: 99%

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Prevalence of Campylobacter hepaticus specific antibodies among commercial free-range layers in Australia

Muralidharan

Huang²,

Anwar³

et al. 2022

Front. Vet. Sci.

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Spotty liver disease (SLD) caused by Campylobacter hepaticus affects the health and productivity of layer hens and is a disease of concern in poultry. In this study, blood and cloacal swab samples were collected from 709 birds across 11 free-range layer farms from different regions of Australia. The prevalence of C. hepaticus specific antibodies and DNA was assessed using a C. hepaticus specific ELISA and PCR and its correlation with mortalities and changes in egg production was analyzed to better understand the seroprevalence of C. hepaticus in Australian free-range layer farms. C. hepaticus specific antibodies were detected from birds in four of the five farms that had no history of SLD with seroprevalence as high as 41% in one of the farms. Seroprevalence of anti-C. hepaticus antibodies among flocks that had an active or previous SLD outbreak varied between 2 and 64%. C. hepaticus DNA was detected from birds in three farms with no known SLD history and five farms with confirmed SLD outbreaks. A good correlation was observed between the ELISA and PCR results with a Pearson correlation coefficient value of 0.85 (p-value = 0.001). No correlation was observed between the flock size or flock age and ELISA or PCR outcomes, and no significant difference between the seroprevalence of anti-C. hepaticus antibodies among flocks with or without a known history of SLD was established (p = 0.143). This study demonstrates the usefulness of C. hepaticus specific ELISA and PCR in identifying the occurrence of mild or sub-clinical SLD and provides a broader and more complete understanding of SLD epidemiology that will inform future research aimed at the development of methods to control SLD, such as appropriate biosecurity measures, vaccines, and feed additives.

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Section: Introductionmentioning

confidence: 96%

Section: Introductionmentioning

confidence: 99%

Prevalence of Campylobacter hepaticus specific antibodies among commercial free-range layers in Australia

Muralidharan

Huang²,

Anwar³

et al. 2022

Front. Vet. Sci.

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“…Pre-absorption removes cross-reactive antibodies from bird sera that would otherwise have bound to C. hepaticus proteins and resulted in false positive readings. The successful differentiation of SLD positive and negative sera samples using SLD-ELISA1 indicated the presence of C. hepaticus specific immunogenic proteins that are not cross reactive with C. jejuni antibodies commonly present in bird sera ( 10 ). Although the assay was highly specific and sensitive, there were a few shortcomings that made it less than ideal for large scale use.…”

Section: Introductionmentioning

confidence: 99%

The use of filamentous hemagglutinin adhesin to detect immune responses to Campylobacter hepaticus infections in layer hens

Muralidharan

Quinteros²,

Anwar³

et al. 2022

Front. Vet. Sci.

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Campylobacter hepaticus is the aetiological agent of Spotty Liver Disease (SLD). SLD can cause significant production loss and mortalities among layer hens at and around peak of lay. We previously developed an enzyme linked immunosorbent assay (ELISA), SLD-ELISA1, to detect C. hepaticus specific antibodies from bird sera using C. hepaticus total proteins and sera pre-absorbed with Campylobacter jejuni proteins. The high specificity achieved with SLD-ELISA1 indicated the presence of C. hepaticus specific antibodies in sera of infected birds. However, some of the reagents used in SLD-ELISA1 are time consuming to prepare and difficult to quality control. This understanding led to the search for C. hepaticus specific immunogenic proteins that could be used in recombinant forms as antibody capture antigens in immunoassay design. In this study, an immunoproteomic approach that combined bioinformatics analysis, western blotting, and LC MS/MS protein profiling was used, and a fragment of filamentous hemagglutinin adhesin (FHA), FHA1,628−1,899 with C. hepaticus specific antigenicity was identified. Recombinant FHA1,628−1,899 was used as antigen coating on ELISA plates to capture FHA1,628−1,899 specific antibodies in sera of infected birds. SLD-ELISA2, based on the purified recombinant FHA fragment, is more user-friendly and standardizable than SLD-ELISA1 for screening antibody responses to C. hepaticus exposure in hens. This study is the first report of the use of FHA from a Campylobacter species in immunoassays, and it also opens future research directions to investigate the role of FHA in C. hepaticus pathogenesis and its effectiveness as a vaccine candidate.

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“…2,3 However, currently available methods for Salmonella detection are subject to some limitations, such as long time (Culture), 4 complex nucleic acid extraction (polymerase chain reaction, PCR), 5 and insufficient sensitivity (enzyme-linked immunosorbent assay, ELISA). 6 Hence, in-field, fast, and sensitive methods to detect Salmonella in foods are urgently needed.…”

Section: Introductionmentioning

confidence: 99%

“…The World Health Organization (WHO) estimated that nearly 1 in 10 people suffer from foodborne diseases in the world each year, and 33 million of healthy life years are lost annually . Among foodborne pathogens, the infections caused by Salmonella is the highest, and Salmonella is one of the four main global causes of diarrhea. , However, currently available methods for Salmonella detection are subject to some limitations, such as long time (Culture), complex nucleic acid extraction (polymerase chain reaction, PCR), and insufficient sensitivity (enzyme-linked immunosorbent assay, ELISA) . Hence, in-field, fast, and sensitive methods to detect Salmonella in foods are urgently needed.…”

Section: Introductionmentioning

confidence: 99%

Fe-MIL-88NH₂ Metal–Organic Framework Nanocubes Decorated with Pt Nanoparticles for the Detection of Salmonella

Guo

Cai

et al. 2021

ACS Appl. Nano Mater.

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Early screening of Salmonella is of utmost importance to minimize the incidence of related foodborne diseases. Here, we reported an immunosensor to fast detect Salmonella using Fe-MIL-88NH2 metal–organic framework (Fe-MOF) nanocubes to load platinum nanoparticles (PtNPs) for signal amplification and smartphone thermal imaging to monitor the temperature change. The Fe-MOF nanocubes decorated with PtNPs (Fe-MOF/PtNPs) were prepared as peroxidase mimics to catalyze hydrogen peroxide (H2O2) to produce O2. The immune magnetic nanoparticles (MNPs), the target bacteria, and the immune Fe-MOF/PtNPs were first mixed by the three-dimensional (3D) chaotic micromixer and conjugated in the microfluidic chip to form MNP–bacteria–Fe-MOF/PtNPs sandwich complexes. After magnetic separation and washing, H2O2 was injected and decomposed by Fe-MOF/PtNPs on the sandwich complexes to generate O2 in a sealed chamber. The increase in the pressure pushed the preloaded water to react with calcium oxide powder, resulting in the generation of heat, which was measured using the smartphone-based thermal sensor to determine the bacteria. The microfluidic immunosensor could detect Salmonella as low as 93 CFU/mL within 1 h.

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Development of an enzyme-linked immunosorbent assay for detecting Campylobacter hepaticus specific antibodies in chicken sera – a key tool in Spotty Liver Disease screening and vaccine development

Abstract: Van (2020): Development of an enzyme-linked immunosorbent assay for detecting Campylobacterhepaticus specific antibodies in chicken sera-a key tool in Spotty Liver Disease screening and vaccine development, Avian Pathology,

Cited by 8 publications

References 22 publications

Prevalence of Campylobacter hepaticus specific antibodies among commercial free-range layers in Australia

Prevalence of Campylobacter hepaticus specific antibodies among commercial free-range layers in Australia

The use of filamentous hemagglutinin adhesin to detect immune responses to Campylobacter hepaticus infections in layer hens

Fe-MIL-88NH₂ Metal–Organic Framework Nanocubes Decorated with Pt Nanoparticles for the Detection of Salmonella

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Development of an enzyme-linked immunosorbent assay for detecting Campylobacter hepaticus specific antibodies in chicken sera – a key tool in Spotty Liver Disease screening and vaccine development

Abstract: Van (2020): Development of an enzyme-linked immunosorbent assay for detecting Campylobacterhepaticus specific antibodies in chicken sera-a key tool in Spotty Liver Disease screening and vaccine development, Avian Pathology,

Cited by 8 publications

References 22 publications

Prevalence of Campylobacter hepaticus specific antibodies among commercial free-range layers in Australia

Prevalence of Campylobacter hepaticus specific antibodies among commercial free-range layers in Australia

The use of filamentous hemagglutinin adhesin to detect immune responses to Campylobacter hepaticus infections in layer hens

Fe-MIL-88NH2 Metal–Organic Framework Nanocubes Decorated with Pt Nanoparticles for the Detection of Salmonella

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Product

Resources

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Fe-MIL-88NH₂ Metal–Organic Framework Nanocubes Decorated with Pt Nanoparticles for the Detection of Salmonella