New Route for Fast Detection of Antibodies against Zoonotic Pathogens in Sera of Slaughtered Pigs by Means of Flow-through Chemiluminescence Immunochips

Wutz, Klaus; Meyer, Verena Katharina; Wacheck, Silke; Krol, P.; Gareis, M.; Nölting, Christina; Struck, Friedhelm; Soutschek, Erwin; Böcher, Oliver; Seidel, Michael

doi:10.1021/ac400781t

Cited by 15 publications

(11 citation statements)

References 49 publications

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“…However, traditional microarray detection requires expensive fluorescence scanners, which limit its use. Analytical CL is a versatile, sensitive method of detection with a wide range of uses, including enzyme-linked immunosorbent assays (Liu et al 2012; Maiolini et al 2013), lateral flow immunoassays (Maiolini et al 2013; Wolter et al 2008; Wutz et al 2013), flow-through hybridization assays (Hommatsu et al 2013), capillary electrophoresis (Jiang et al 2013), flow injection CL analysis (Tan and Song 2013), and magnetic bead-based DNA hybridization assays (Li and He 2009). This is the first paper to subtype influenza viruses using a chemiluminescent oligonucleotide microarray.…”

Section: Discussionmentioning

confidence: 99%

Genotyping and detection of common avian and human origin-influenza viruses using a portable chemiluminescence imaging microarray

et al. 2016

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Background Influenza viruses are divided into three types, A, B, and C. Human influenza A and B viruses can cause seasonal epidemics, but influenza C causes only a mild respiratory illness. Influenza A virus can infect various host species. In 2013, human-infectious avian influenza A (H7N9) was first reported in China. By the second week of 2014, there were 210 laboratory-confirmed human cases in the country, and the mortality rate eventually reached 22 %. Rapid and accurate diagnosis of influenza viruses is important for clinical management and epidemiology. MethodsIn this assay, a cost-effective chemiluminescence (CL) detection oligonucleotide microarray was developed to genotype and detect avian influenza A (H7N9), avian influenza A (H5N1), 2009 influenza A (H1N1), seasonal influenza A (H1N1), and seasonal influenza A (H3N2). Influenza A viruses and influenza B viruses were also generally detected using this microarray.ResultsThe results of detection of 40 cultivated influenza virus strains showed that the microarray was able to distinguish the subtypes of these influenza viruses very well. The microarray possessed similar or 10 fold higher limit of detection than the real-time RT-PCR method. Sixty-six clinical swab samples were detected using this microarray and verified with real time RT-PCR to evaluate the efficiency of this microarray for clinical testing.ConclusionsA reliable CL detection oligonucleotide microarray had been developed to genotype and detected these influenza viruses.

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Section: Discussionmentioning

confidence: 99%

Genotyping and detection of common avian and human origin-influenza viruses using a portable chemiluminescence imaging microarray

et al. 2016

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“…Sufficient amounts of meat juice can be obtained from a 10‐g piece of muscle and frozen for later use, for example in animal disease surveys. Novel automated technologies may even further facilitate the future use of serological tests (Wutz et al., ).…”

Section: Discussionmentioning

confidence: 99%

Meat Juice Serology and Improved Food Chain Information as Control Tools for Pork‐Related Public Health Hazards

Felin

Jukola

Raulo

et al. 2014

Zoonoses and Public Health

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The seroprevalence of Salmonella spp., pathogenic Yersinia spp., Toxoplasma gondii and Trichinella spp. was studied in 1353 finishing pigs from 259 farms that were allocated according to farm types: large fattening farms (≥ 1000 pig places), small fattening farms (< 1000 pig places) and farrow-to-finish farms. The antibodies were analysed with commercial ELISA kits in meat juice samples that were collected at Finnish slaughterhouses. Salmonella antibodies were rare (3% of pigs, 14% of farms) when the cut-off optical density (OD) value 0.2 was used. Antibodies to pathogenic Yersinia spp. and T. gondii were detected in 57% of pigs and 85% of farms (OD ≥ 0.3) and in 3% of pigs and 9% of farms (OD ≥ 0.15), respectively. No antibodies to Trichinella spp. were detected (OD ≥ 0.3). The European Food Safety Authority (EFSA) considers Salmonella spp., Yersinia enterocolitica, T. gondii and Trichinella spp. as the most relevant biological hazards in the context of meat inspection of pigs. The seroprevalence of these important zoonotic pathogens was low in Finland, except that of Yersinia. The seroprevalence of Toxoplasma was significantly higher in pigs originating from small-scale fattening farms (P < 0.05). Strong positive correlation was observed at the animal level between Salmonella and Yersinia seropositivity and between Salmonella and Toxoplasma seropositivity (P < 0.05). We suggest that these results reflect the level and importance of biosecurity measures applied on the farms. Meat juice serology at slaughter is a useful tool for targeting measures to control these pathogens. The information obtained from analyses should be used as part of the food chain information (FCI).

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“…The preparation process of rAg microarrays was performed as described elsewhere [ 24 ]. The recombinant antigens of HEV and Yersinia spp.…”

Section: Methodsmentioning

confidence: 99%

“…The measurement program for the analysis of antibodies in sera of slaughtered pigs is described elsewhere in detail [ 24 ]. The microarray chip was placed in the detection unit of the MCR3 from GWK Präzisionstechnik GmbH (Munich, Germany) and was connected with the flow system by closing the upper part of the detection unit (see Figure 1 ).…”

Section: Methodsmentioning

confidence: 99%

“…An immunochip was developed that is able to detect antibodies against emergent zoonotic pathogens like Yersinia spp. and HEV in swine sera by affinity binding to recombinant antigens [ 24 ]. On the MCR3, the regeneration of microarray chips has only been demonstrated so far for indirect competitive microarray immunoassays [ 25 ] with small organic molecules like antibiotics [ 26 ], phycotoxins [ 27 ], mycotoxins [ 28 ], or carbohydrates [ 29 ] immobilized on the surface.…”

Section: Introductionmentioning

confidence: 99%

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Regeneration of Recombinant Antigen Microarrays for the Automated Monitoring of Antibodies against Zoonotic Pathogens in Swine Sera

Meyer

Kober

Seidel

2015

Sensors

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The ability to regenerate immobilized proteins like recombinant antigens (rAgs) on surfaces is an unsolved problem for flow-based immunoassays on microarray analysis systems. The regeneration on microarray chip surfaces is achieved by changing the protein structures and desorption of antibodies. Afterwards, reactivation of immobilized protein antigens is necessary for reconstitution processes. Any backfolding should be managed in a way that antibodies are able to detect the protein antigens in the next measurement cycle. The regeneration of rAg microarrays was examined for the first time on the MCR3 flow-based chemiluminescence (CL) microarray analysis platform. The aim was to reuse rAg microarray chips in order to reduce the screening effort and costs. An antibody capturing format was used to detect antibodies against zoonotic pathogens in sera of slaughtered pigs. Different denaturation and reactivation buffers were tested. Acidic glycine-SDS buffer (pH 2.5) and 8 M guanidinium hydrochloride showed the best results in respect of denaturation efficiencies. The highest CL signals after regeneration were achieved with a carbonate buffer containing 10 mM DTT and 0.1% BSA for reactivation. Antibodies against Yersinia spp. and hepatitis E virus (HEV) were detected in swine sera on one immunochip over 4 days and 25 measurement cycles. Each cycle took 10 min for detection and regeneration. By using the rAg microarray chip, a fast and automated screening of antibodies against pathogens in sera of slaughtered pigs would be possible for zoonosis monitoring.

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New Route for Fast Detection of Antibodies against Zoonotic Pathogens in Sera of Slaughtered Pigs by Means of Flow-through Chemiluminescence Immunochips

Cited by 15 publications

References 49 publications

Genotyping and detection of common avian and human origin-influenza viruses using a portable chemiluminescence imaging microarray

Genotyping and detection of common avian and human origin-influenza viruses using a portable chemiluminescence imaging microarray

Meat Juice Serology and Improved Food Chain Information as Control Tools for Pork‐Related Public Health Hazards

Regeneration of Recombinant Antigen Microarrays for the Automated Monitoring of Antibodies against Zoonotic Pathogens in Swine Sera

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