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Tanja Opriessnig - One of the best experts on this subject based on the ideXlab platform.

  • Erysipelothrix rhusiopathiae serotype 15 associated with recurring pig erysipelas outbreaks
    Veterinary Record, 2018
    Co-Authors: Priscilla F Gerber, Tanja Opriessnig, Alasdair Macleod
    Abstract:

    Erysipelothrix rhusiopathiae is the causative agent of pig erysipelas and can be associated with sporadic cases or larger outbreaks of septicaemia with characteristic skin lesions or chronic polyarthritis.1 Within the genus Erysipelothrix , at least 6 species ( Erysipelothrix rhusiopathiae , Erysipelothrix tonsillarum , Erysipelothrix species strain 1, Erysipelothrix species strain 2, Erysipelothrix species strain 3 and Erysipelothrix inopinata ) and 28 serotypes (1a, 1b, 2–26 and N) have been recognised.1 E rhusiopathiae serotypes 1 and 2 are frequently isolated from clinically affected pigs, although other E rhusiopathiae serotypes have been sporadically associated with clinical disease.1 2 While there is no experimental evidence that Erysipelothrix species other than E rhusiopathiae cause disease in pigs,3 certain Erysipelothrix species strains have been isolated from clinical cases4 5 and from condemned carcases in abattoirs.2 6 Pig erysipelas is generally seen in adults and grow-finish pigs after the decline of maternal antibodies.1 Humoural immunity is considered most important for disease prevention and vaccines containing live or inactivated E rhusiopathiae serotype 1 or 2 isolates are commonly used.7 In the UK, there are two E rhusiopathiae bacterins available commercially based on serotype 2 or serotypes 1 and 2.8 In recent years, the incidence of E rhusiopathiae infection in pigs appears to have increased worldwide2 9–11 and is also increasing in European poultry production systems.12 This study summarises the findings associated with chronic E rhusiopathiae infection in a commercial wean-finish pig herd in the UK (farm A) that received piglets from an E rhusiopathiae vaccinated high health breeding herd free of porcine reproductive and respiratory syndrome virus (PRRSV) and Mycoplasma hyopneumoniae as monitored by serological testing in three-month intervals. Specifically, the breeding farm used a commercial E rhusiopathiae serotype 2-based bacterin (Porcillis Ery, Intervet UK) which was administered …

  • serotypes and spa types of Erysipelothrix rhusiopathiae isolates from british pigs 1987 to 2015
    Veterinary Journal, 2017
    Co-Authors: Mark Mcneil, Priscilla F Gerber, Jill R Thomson, Susanna Williamson, Tanja Opriessnig
    Abstract:

    Erysipelothrix spp. cause a range of clinical signs in pigs and at least 28 different Erysipelothrix spp. serotypes have been identified. In this study, 128 isolates of Erysipelothrix spp. from pigs in Great Britain from 1987 to 2015 were characterised by serotyping and multiplex real time PCR assays targeting the surface protective antigen (Spa) and the main genotypes (Erysipelothrix rhusiopathiae, Erysipelothrix tonsillarum and Erysipelothrix spp. strain 2). All 128 British isolates were characterised as E. rhusiopathiae and were classified as serotypes 1a (n=21), 1b (n=17), 2 (n=75), 5 (n=2), 9 (n=2), 10 (n=2), 11 (n=4) and 15 (n=1), while four isolates were untypeable. All isolates were positive for the spa A gene. Serotypes 1a, 1b and 2 constituted 88.3% of the isolates; current serotype 2 based vaccines should protect against these isolates.

  • Additional file 2: of Genomic analysis of the multi-host pathogen Erysipelothrix rhusiopathiae reveals extensive recombination as well as the existence of three generalist clades with wide geographic distribution
    2016
    Co-Authors: Taya Forde, Tanja Opriessnig, Roman Biek, Ruth Zadoks, Matthew Workentine, Jeroen De Buck, Susan Kutz, Hannah Trewby, Frank Van Der Meer, Karin Orsel
    Abstract:

    Supplementary figures. Figure S1. Erysipelothrix rhusiopathiae core genome. Core genes were plotted against the E. rhusiopathiae Fujisawa reference genome using GView, filtering out low-complexity sequences (e.g. repetitive regions). Bacteriophage sequences in the annotated reference genome and core genes associated with bacterial competence are highlighted. Publicly available E. rhusiopathiae isolates and de novo assembled isolates whose average depth of coverage was greater than 15X were included in this analysis. Figure S2. Recombinant fragments estimated in the Erysipelothrix rhusiopathiae core genome using BratNextGen. Presence of the same color block across multiple isolates within a column represents acquisition of the same recombinant segment; otherwise colors are arbitrary. Figure S3. Recombinant fragments in Clades 1, 2 and 3 respectively, estimated in Gubbins. Red blocks are recombinant fragments that have been inherited by multiple isolates, while blue fragments are unique to that isolate. Figure S4. Population subgroup assignment of Erysipelothrix rhusiopathiae isolates during chromosome painting using ChromoPainter/fineSTRUCTURE as shown in Fig. 4. Figure S5. Homoplasy associated with Erysipelothrix rhusiopathiae serotyping. This figure shows examples of homoplasy in serotypes 1a, 2 and 5, based on the same phylogenetic tree shown in Fig. 3. Labelled arrows show the locations of isolates of the different serotypes. Serotype was previously determined for all isolates shown here except VI11-2_Lu, which was serotyped in this study. Figure S6. Data analysis pipeline. (PDF 695 kb

  • Additional file 1: Table S1. of Genomic analysis of the multi-host pathogen Erysipelothrix rhusiopathiae reveals extensive recombination as well as the existence of three generalist clades with wide geographic distribution
    2016
    Co-Authors: Taya Forde, Tanja Opriessnig, Roman Biek, Ruth Zadoks, Matthew Workentine, Jeroen De Buck, Susan Kutz, Hannah Trewby, Frank Van Der Meer, Karin Orsel
    Abstract:

    Excel spreadsheet of the Erysipelothrix spp. isolates used in this study, including metadata and assembly statistics. (XLSX 24 kb

  • Erysipelothrix rhusiopathiae isolates recovered from fish a harbour seal phoca vitulina and the marine environment are capable of inducing characteristic cutaneous lesions in pigs
    Journal of Comparative Pathology, 2013
    Co-Authors: Tanja Opriessnig, J.s. Bender, Huigang Shen, J R Boehm, Patrick G. Halbur
    Abstract:

    Summary In order to determine the diversity and pathogenicity of Erysipelothrix spp. isolates recovered from marine fish, a harbour seal (Phoca vitulina) and the marine environment, 14 isolates were characterized by genotyping, serotyping, determination of the surface protective antigen (spa) gene type and assessment of virulence in a pig bioassay. All 14 isolates were Erysipelothrix rhusiopathiae. Isolates were determined to be of serotypes 2 (n = 3), 3 (n = 1), 4 (n = 1), 12 (n = 1), 15 (n = 1) or 21 (n = 6), and one isolate cross-reacted with serotypes 5 and 21. The spa gene analysis determined that 64.3% (n = 9) were spaA and 35.7% (n = 5) were spaB1. In pigs, 10/14 isolates induced small plaques to diamond-shaped cutaneous lesions consistent with Erysipelothrix spp. infection. The results of this study indicate that the marine E. rhusiopathiae isolates have greater genetic and antigenic diversity than pig isolates and are capable of inducing classical skin lesions in pigs.

James T Perkins - One of the best experts on this subject based on the ideXlab platform.

  • visceral botryomycosis in a case of Erysipelothrix rhusiopathiae endocarditis
    Human Pathology, 2005
    Co-Authors: Henry J. Carson, James T Perkins
    Abstract:

    Summary Botryomycosis is a rare bacterial infection of the skin and, rarely, viscera that is characterized by the formation of characteristic hyaline grains. We encountered a patient with Erysipelothrix rhusiopathiae endocarditis who developed visceral botryomycosis. He was a 54-year-old black man who presented in sepsis with a history of progressive weakness and severe weight loss. He died 10 hours after admission. At autopsy, endocarditis was identified, along with infarcts of the spleen and kidneys. Microscopically, visceral botryomycosis was seen. With these bacteria, an animal source is usually identified, although one was not found in this man. Erysipelothrix rhusiopathiae is an organism that is becoming identified in a range of infections in humans, the manifestations and pathophysiology of which are still being discovered.

  • visceral botryomycosis in a case of Erysipelothrix rhusiopathiae endocarditis
    Human Pathology, 2005
    Co-Authors: Henry J. Carson, James T Perkins
    Abstract:

    Summary Botryomycosis is a rare bacterial infection of the skin and, rarely, viscera that is characterized by the formation of characteristic hyaline grains. We encountered a patient with Erysipelothrix rhusiopathiae endocarditis who developed visceral botryomycosis. He was a 54-year-old black man who presented in sepsis with a history of progressive weakness and severe weight loss. He died 10 hours after admission. At autopsy, endocarditis was identified, along with infarcts of the spleen and kidneys. Microscopically, visceral botryomycosis was seen. With these bacteria, an animal source is usually identified, although one was not found in this man. Erysipelothrix rhusiopathiae is an organism that is becoming identified in a range of infections in humans, the manifestations and pathophysiology of which are still being discovered.

Henry J. Carson - One of the best experts on this subject based on the ideXlab platform.

  • visceral botryomycosis in a case of Erysipelothrix rhusiopathiae endocarditis
    Human Pathology, 2005
    Co-Authors: Henry J. Carson, James T Perkins
    Abstract:

    Summary Botryomycosis is a rare bacterial infection of the skin and, rarely, viscera that is characterized by the formation of characteristic hyaline grains. We encountered a patient with Erysipelothrix rhusiopathiae endocarditis who developed visceral botryomycosis. He was a 54-year-old black man who presented in sepsis with a history of progressive weakness and severe weight loss. He died 10 hours after admission. At autopsy, endocarditis was identified, along with infarcts of the spleen and kidneys. Microscopically, visceral botryomycosis was seen. With these bacteria, an animal source is usually identified, although one was not found in this man. Erysipelothrix rhusiopathiae is an organism that is becoming identified in a range of infections in humans, the manifestations and pathophysiology of which are still being discovered.

  • visceral botryomycosis in a case of Erysipelothrix rhusiopathiae endocarditis
    Human Pathology, 2005
    Co-Authors: Henry J. Carson, James T Perkins
    Abstract:

    Summary Botryomycosis is a rare bacterial infection of the skin and, rarely, viscera that is characterized by the formation of characteristic hyaline grains. We encountered a patient with Erysipelothrix rhusiopathiae endocarditis who developed visceral botryomycosis. He was a 54-year-old black man who presented in sepsis with a history of progressive weakness and severe weight loss. He died 10 hours after admission. At autopsy, endocarditis was identified, along with infarcts of the spleen and kidneys. Microscopically, visceral botryomycosis was seen. With these bacteria, an animal source is usually identified, although one was not found in this man. Erysipelothrix rhusiopathiae is an organism that is becoming identified in a range of infections in humans, the manifestations and pathophysiology of which are still being discovered.

Eric K Pomaranski - One of the best experts on this subject based on the ideXlab platform.

  • description of Erysipelothrix piscisicarius sp nov an emergent fish pathogen and assessment of virulence using a tiger barb puntigrus tetrazona infection model
    International Journal of Systematic and Evolutionary Microbiology, 2020
    Co-Authors: Eric K Pomaranski, Matt J Griffin, Alvin C Camus, Abigail R Armwood, Johnny Shelley, Geoffrey C Waldbieser, Benjamin R Lafrentz, Julio C Garcia, Roy P E Yanong, Esteban Soto
    Abstract:

    A recently described emergent disease of ornamental fish has been associated with an Erysipelothrix species positive for the surface protective antigen (spa) C gene. Whole genome sequencing was performed on five spaC Erysipelothrix isolates from diseased ornamental fish. In addition, these spaC Erysipelothrix isolates were compared to spaA-, spaB- and other spaC-positive Erysipelothrix species isolated from terrestrial and marine mammals, birds and fish using multi-locus sequence analysis (MLSA). The genomes of fish pathogenic spaC isolates were genetically distinct from Erysipelothrix rhusiopathiae , sharing 86.61–86.94 % average nucleotide identity (ANI) and digital DNA–DNA hybridization (dDDH) values of 31.6–32.2 %, but 99.01–99.11 % ANI and 90.8–91.9 % dDDH values with the uncharacterized spaC-positive Erysipelothrix sp. strain 2 isolated from swine. The findings indicate the spaC-positive fish and swine isolates are conspecific and represent a previously unrecognized taxon. While phylogenies inferred from MLSA sequences confirm this conclusion, slight genetic differences between the spaC fish isolates and swine strain 2 were indicated. Bath immersion challenge trials were conducted using tiger barbs (Puntigrus tetrazona) exposed by immersion to 107 c.f.u. ml−1 of three fish pathogenic spaC Erysipelothrix species, and three spaA and two spaB E. rhusiopathiae isolates as a model of infection. Thirty days post-challenge, cumulative mean percentage survival was 37 % for the spaA, 100 % for the spaB and 13 % for the spaC isolates, revealing differences in virulence among the various spa genotypes in fish. Genetic findings and observed differences in virulence demonstrate the fish pathogenic spaC isolates represent a novel species, for which the name Erysipelothrix piscisicarius sp. nov. is proposed. The type strain is E. piscisicarius 15TAL0474T (=NRRL B-65533T=ATCC-TSD-175T=DSM 110099T).

  • first detection of Erysipelothrix sp infection in western mosquitofish gambusia affinis inhabiting catfish aquaculture ponds in mississippi usa
    Diseases of Aquatic Organisms, 2019
    Co-Authors: Justin M Stilwell, Eric K Pomaranski, Matt J Griffin, Esteban Soto, Thomas G Rosser, John H Leary, Katharina Hagenfrei, Charles C Mischke, Alvin C Camus
    Abstract:

    : Native and introduced fish can serve as reservoirs for pathogens of cultured fish species. In the current study, 351 archived western mosquitofish Gambusia affinis collected from experimental catfish production ponds in Mississippi, USA, were surveyed histologically to evaluate their potential as vectors for fish pathogens. In addition to epitheliocystis and multiple metazoan parasites, 8 fish had widespread basophilic colonies of small Gram-positive rods associated primarily with stroma supporting the skeletal muscle and bone, as well as connective tissue components of other tissues and organ systems, such as perivascular adventitia and basement membranes. These findings were consistent with spaC-type Erysipelothrix sp. infections in ornamental fish cultured in the USA. The 16S rRNA, gyrase B (gyrB), and surface protective antigen (spa) genes were amplified and sequenced from bacterial colonies excised from paraffin-embedded tissue sections using laser capture microdissection. Molecular data confirmed the identity of a spaC-type Erysipelothrix sp., which grouped phylogenetically with spaC-type Erysipelothrix sp. from diseased ornamental fish. Given the significance of commercial catfish aquaculture in the southeastern USA and the widespread distribution of mosquitofish in catfish ponds throughout the region, infectivity trials with channel catfish Ictalurus punctatus were conducted. Catfish fingerlings were exposed to a spaC-type Erysipelothrix sp. isolate by intracoelomic injection and gavage. No mortality was observed in catfish exposed by either route, and surviving fish demonstrated no significant histopathologic lesions, suggesting channel catfish have low susceptibility to the bacteria. Further research is warranted to investigate the susceptibility of other cultured fish species to this emergent fish pathogen.

  • characterization of spac type Erysipelothrix sp isolates causing systemic disease in ornamental fish
    Journal of Fish Diseases, 2018
    Co-Authors: Eric K Pomaranski, Roy P E Yanong, Stephen R Reichley, John P Shelley, Deborah B Pouder, J C Wolf, Kirsten V Kenelty, B Van Bonn, F Oliaro, Barbara A Byrne
    Abstract:

    Since 2012, low-to-moderate mortality associated with an Erysipelothrix sp. bacterium has been reported in ornamental fish. Histological findings have included facial cellulitis, necrotizing dermatitis and myositis, and disseminated coelomitis with abundant intralesional Gram-positive bacterial colonies. Sixteen Erysipelothrix sp. isolates identified phenotypically as E. rhusiopathiae were recovered from diseased cyprinid and characid fish. Similar clinical and histological changes were also observed in zebrafish, Danio rerio, challenged by intracoelomic injection. The Erysipelothrix sp. isolates from ornamental fish were compared phenotypically and genetically to E. rhusiopathiae and E. tonsillarum isolates recovered from aquatic and terrestrial animals from multiple facilities. Results demonstrated that isolates from diseased fish were largely clonal and divergent from E. rhusiopathiae and E. tonsillarum isolates from normal fish skin, marine mammals and terrestrial animals. All ornamental fish isolates were PCR positive for spaC, with marked genetic divergence (<92% similarity at gyrB, <60% similarity by rep-PCR) between the ornamental fish isolates and other Erysipelothrix spp. isolates. This study supports previous work citing the genetic variability of Erysipelothrix spp. spa types and suggests isolates from diseased ornamental fish may represent a genetically distinct species.

  • Characterization of spaC-type Erysipelothrix sp. isolates causing systemic disease in ornamental fish.
    Journal of fish diseases, 2017
    Co-Authors: Eric K Pomaranski, Roy P E Yanong, Stephen R Reichley, John P Shelley, Deborah B Pouder, J C Wolf, Kirsten V Kenelty, B Van Bonn, F Oliaro, Barbara A Byrne
    Abstract:

    Since 2012, low-to-moderate mortality associated with an Erysipelothrix sp. bacterium has been reported in ornamental fish. Histological findings have included facial cellulitis, necrotizing dermatitis and myositis, and disseminated coelomitis with abundant intralesional Gram-positive bacterial colonies. Sixteen Erysipelothrix sp. isolates identified phenotypically as E. rhusiopathiae were recovered from diseased cyprinid and characid fish. Similar clinical and histological changes were also observed in zebrafish, Danio rerio, challenged by intracoelomic injection. The Erysipelothrix sp. isolates from ornamental fish were compared phenotypically and genetically to E. rhusiopathiae and E. tonsillarum isolates recovered from aquatic and terrestrial animals from multiple facilities. Results demonstrated that isolates from diseased fish were largely clonal and divergent from E. rhusiopathiae and E. tonsillarum isolates from normal fish skin, marine mammals and terrestrial animals. All ornamental fish isolates were PCR positive for spaC, with marked genetic divergence (

J.s. Bender - One of the best experts on this subject based on the ideXlab platform.

  • Erysipelothrix rhusiopathiae isolates recovered from fish a harbour seal phoca vitulina and the marine environment are capable of inducing characteristic cutaneous lesions in pigs
    Journal of Comparative Pathology, 2013
    Co-Authors: Tanja Opriessnig, J.s. Bender, Huigang Shen, J R Boehm, Patrick G. Halbur
    Abstract:

    Summary In order to determine the diversity and pathogenicity of Erysipelothrix spp. isolates recovered from marine fish, a harbour seal (Phoca vitulina) and the marine environment, 14 isolates were characterized by genotyping, serotyping, determination of the surface protective antigen (spa) gene type and assessment of virulence in a pig bioassay. All 14 isolates were Erysipelothrix rhusiopathiae. Isolates were determined to be of serotypes 2 (n = 3), 3 (n = 1), 4 (n = 1), 12 (n = 1), 15 (n = 1) or 21 (n = 6), and one isolate cross-reacted with serotypes 5 and 21. The spa gene analysis determined that 64.3% (n = 9) were spaA and 35.7% (n = 5) were spaB1. In pigs, 10/14 isolates induced small plaques to diamond-shaped cutaneous lesions consistent with Erysipelothrix spp. infection. The results of this study indicate that the marine E. rhusiopathiae isolates have greater genetic and antigenic diversity than pig isolates and are capable of inducing classical skin lesions in pigs.

  • characterization of Erysipelothrix species isolates from clinically affected pigs environmental samples and vaccine strains from six recent swine erysipelas outbreaks in the united states
    Clinical and Vaccine Immunology, 2010
    Co-Authors: J.s. Bender, Huigang Shen, Christa Irwin, Kent J Schwartz, Tanja Opriessnig
    Abstract:

    The aim of this study was to characterize Erysipelothrix sp. isolates from clinically affected pigs and their environment and compare them to the Erysipelothrix sp. vaccines used at the sites. Samples were collected during swine erysipelas outbreaks in vaccinated pigs in six Midwest United States swine operations from 2007 to 2009. Pig tissue samples were collected from 1 to 3 pigs from each site. Environmental samples (manure, feed, central-line water, oral fluids, and swabs collected from walls, feed lines, air inlets, exhaust fans, and nipple drinkers) and live vaccine samples were collected following the isolation of Erysipelothrix spp. from clinically affected pigs. All Erysipelothrix sp. isolates obtained were further characterized by serotyping. Selected isolates were further characterized by PCR assays for genotype (E. rhusiopathiae, E. tonsillarum, Erysipelothrix sp. strain 1, and Erysipelothrix sp. strain 2) and surface protective antigen (spa) type (A, B1, B2, and C). All 26 isolates obtained from affected pigs were E. rhusiopathiae, specifically, serotypes 1a, 1b, 2, and 21. From environmental samples, 56 isolates were obtained and 52/56 were E. rhusiopathiae (serotypes 1a, 1b, 2, 6, 9, 12, and 21), 3/56 were Erysipelothrix sp. strain 1 (serotypes 13 and untypeable), and one was a novel species designated Erysipelothrix sp. strain 3 (serotype untypeable). Four of six vaccines used at the sites were commercially available products and contained live E. rhusiopathiae serotype 1a. Of the remaining two vaccines, one was an autogenous live vaccine and contained live E. rhusiopathiae serotype 2 and one was a commercially produced inactivated vaccine and was described by the manufacturer to contain serotype 2 antigen. All E. rhusiopathiae isolates were positive for spaA. All Erysipelothrix sp. strain 1 isolates and the novel Erysipelothrix sp. strain 3 isolate were negative for all currently known spa types (A, B1, B2, and C). These results indicate that Erysipelothrix spp. can be isolated from the environment of clinically affected pigs; however, the identified serotypes in pigs differ from those in the environment at the selected sites. At one of the six affected sites, the vaccine strain and the isolates from clinically affected pigs were of homologous serotype; however, vaccinal and clinical isolates were of heterologous serotype at the remaining five sites, suggesting that reevaluation of vaccine efficacy using recent field strains may be warranted.

  • identification of surface protective antigen spa types in Erysipelothrix reference strains and diagnostic samples by spa multiplex real time and conventional pcr assays
    Journal of Applied Microbiology, 2010
    Co-Authors: Huigang Shen, J.s. Bender, Tanja Opriessnig
    Abstract:

    Aim:  To develop spa multiplex real-time and conventional PCR assays to detect and differentiate between spaA, spaB and spaC genes within Erysipelothrix spp. Methods and Results:  For evaluation of the assays, 28 Erysipelothrix spp. reference strains, 25 tissues from pigs inoculated with reference strains of serotypes 1, 2, 5, 10 or 18, and 15 diagnostic samples were used. SpaA was found to be present in Erysipelothrix rhusiopathiae serotypes 1a, 1b, 2, 5, 9, 12, 15, 16, 17, 23 and N; spaB was detected in E. rhusiopathiae serotypes 4, 6, 8, 11, 19 and 21 and spaC was detected in E. sp. strain 2 serotype 18. Spa-related genes were not detected in E. tonsillarum strains (serotypes 3, 7, 10, 14, 20, 22, 24, 25, 26) or E. sp. strain 1 (serotype 13). With the spa multiplex real-time PCR assay, it was also possible to further differentiate spaB into spaB1 (serotypes 4, 6, 8, 19 and 21) and spaB2 (serotype 11). Overall, spaA was detected in seven experimental tissue samples and six diagnostic tissue samples, and spaC in two experimental tissue samples. The detection limits were determined to be five colony-forming units (CFU) per reaction for the spa multiplex real-time PCR assay and 4000 CFU per reaction for the conventional PCR assay. Conclusions:  Both spa PCR assays were specific and reproducible in the identification of spa types in Erysipelothrix spp. Significance and Impact of the Study:  The described spa PCR assays may be useful tools for investigating spa prevalence among strains isolated from field tissues and to determine the role of the Spa proteins in vaccine protection and pathogenesis.

  • rapid detection and differentiation of Erysipelothrix spp by a novel multiplex real time pcr assay
    Journal of Applied Microbiology, 2010
    Co-Authors: Narinder Pal, J.s. Bender, Tanja Opriessnig
    Abstract:

    Aim:  The aim of this study was to develop a multiplex real-time PCR assay for the identification and discrimination of Erysipelothrix rhusiopathiae, tonsillarum and Erysipelothrix sp. strain 2 for direct detection of Erysipelothrix spp. from animal specimens. Methods and Results:  A primer set and three species-specific probes with different end labelling were designed from the noncoding region downstream of the 5S rRNA coding region. The sensitivity, specificity and repeatability of the assay were validated by analysing 27 Erysipelothrix spp. reference serotype strains and ten septicemia-associated non-Erysipelothrix spp. bacterial isolates. Cross-reactivity with Erysipelothrix sp. strain 1 was not observed with any of the primer probe combinations. The detection limit was determined to be <10 colony forming units and as low as one genome equivalent per PCR . Further evaluation of the Erysipelothrix spp. multiplex PCR was performed by comparing an enrichment isolation culture method and a conventional differential PCR on 15 samples from pigs experimentally inoculated with Erysipelothrix spp. and 22 samples from pigs with suspected natural infection. Conclusion:  The multiplex real-time PCR assay was found to be simple, rapid, reliable, specific and highly sensitive. Significance and Impact of the Study:  The developed real-time multiplex PCR assay does not require cumbersome and lengthy cultivation steps prior to DNA extraction, obtained comparable results to enrichment isolation, and will be useful in diagnostic laboratories for rapid detection of Erysipelothrix spp.

  • Rapid detection and differentiation of Erysipelothrix spp. by a novel multiplex real‐time PCR assay
    Journal of applied microbiology, 2009
    Co-Authors: Narinder Pal, J.s. Bender, Tanja Opriessnig
    Abstract:

    Aim:  The aim of this study was to develop a multiplex real-time PCR assay for the identification and discrimination of Erysipelothrix rhusiopathiae, tonsillarum and Erysipelothrix sp. strain 2 for direct detection of Erysipelothrix spp. from animal specimens. Methods and Results:  A primer set and three species-specific probes with different end labelling were designed from the noncoding region downstream of the 5S rRNA coding region. The sensitivity, specificity and repeatability of the assay were validated by analysing 27 Erysipelothrix spp. reference serotype strains and ten septicemia-associated non-Erysipelothrix spp. bacterial isolates. Cross-reactivity with Erysipelothrix sp. strain 1 was not observed with any of the primer probe combinations. The detection limit was determined to be