Novel multiplex PCR approach for the detection of Erysipelothrix rhusiopathiae, Streptococcus suis, and Staphylococcus hyicus in swine
Erysipelothrix rhusiopathiae, Streptococcus suis, and Staphylococcus hyicus are neglected, emerging and zoonotic pathogens in pigs that share common clinical signs and lesions, particularly septicaemia and skin lesions, highlighting the need for differential diagnosis. Currently, no single diagnosti...
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| Language: | English |
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Frontiers Media S.A.
2025-06-01
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| Series: | Frontiers in Veterinary Science |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fvets.2025.1605316/full |
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| author | A. Arun Prince Milton Sabia Khan Kandhan Srinivas Torik Basar Zakir Hussain G. Bhuvana Priya M. Saminathan L. H. Puii Sandeep Ghatak Samir Das |
| author_facet | A. Arun Prince Milton Sabia Khan Kandhan Srinivas Torik Basar Zakir Hussain G. Bhuvana Priya M. Saminathan L. H. Puii Sandeep Ghatak Samir Das |
| author_sort | A. Arun Prince Milton |
| collection | DOAJ |
| description | Erysipelothrix rhusiopathiae, Streptococcus suis, and Staphylococcus hyicus are neglected, emerging and zoonotic pathogens in pigs that share common clinical signs and lesions, particularly septicaemia and skin lesions, highlighting the need for differential diagnosis. Currently, no single diagnostic assay is available to test samples for confirmation of the presence of all three pathogens. To address this, a novel multiplex PCR-based detection assay was optimized and evaluated for the simultaneous detection of E. rhusiopathiae, S. suis, and S. hyicus targeting the, 23S rRNA, recN, and sodA genes, respectively. The optimized multiplex PCR demonstrated 100% analytical specificity, with a detection limit of 104 copies/μL for E. rhusiopathiae and S. hyicus, and 106 copies/μL for S. suis. The assay was further validated by comparison with previously established uniplex PCR assays and tested on field-collected pig blood samples to assess real-world applicability. The developed assay is rapid, specific and cost-effective and can serve as a handy diagnostic tool for testing suspect samples for these neglected bacterial pathogens of pigs. |
| format | Article |
| id | doaj-art-8dcd7c899c3349848d015bd401a06b66 |
| institution | Kabale University |
| issn | 2297-1769 |
| language | English |
| publishDate | 2025-06-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Veterinary Science |
| spelling | doaj-art-8dcd7c899c3349848d015bd401a06b662025-08-20T03:30:19ZengFrontiers Media S.A.Frontiers in Veterinary Science2297-17692025-06-011210.3389/fvets.2025.16053161605316Novel multiplex PCR approach for the detection of Erysipelothrix rhusiopathiae, Streptococcus suis, and Staphylococcus hyicus in swineA. Arun Prince Milton0Sabia Khan1Kandhan Srinivas2Torik Basar3Zakir Hussain4G. Bhuvana Priya5M. Saminathan6L. H. Puii7Sandeep Ghatak8Samir Das9Division of Animal and Fisheries Sciences, ICAR Research Complex for NEH Region, Umiam, IndiaDivision of Animal and Fisheries Sciences, ICAR Research Complex for NEH Region, Umiam, IndiaDivision of Animal and Fisheries Sciences, ICAR Research Complex for NEH Region, Umiam, IndiaDivision of Animal and Fisheries Sciences, ICAR Research Complex for NEH Region, Umiam, IndiaDivision of Animal and Fisheries Sciences, ICAR Research Complex for NEH Region, Umiam, IndiaCollege of Agriculture, Central Agricultural University (Imphal), Kyrdemkulai, IndiaICAR Indian Veterinary Research Institute, Bareilly, IndiaDivision of Animal and Fisheries Sciences, ICAR Research Complex for NEH Region, Umiam, IndiaDivision of Animal and Fisheries Sciences, ICAR Research Complex for NEH Region, Umiam, IndiaDivision of Animal and Fisheries Sciences, ICAR Research Complex for NEH Region, Umiam, IndiaErysipelothrix rhusiopathiae, Streptococcus suis, and Staphylococcus hyicus are neglected, emerging and zoonotic pathogens in pigs that share common clinical signs and lesions, particularly septicaemia and skin lesions, highlighting the need for differential diagnosis. Currently, no single diagnostic assay is available to test samples for confirmation of the presence of all three pathogens. To address this, a novel multiplex PCR-based detection assay was optimized and evaluated for the simultaneous detection of E. rhusiopathiae, S. suis, and S. hyicus targeting the, 23S rRNA, recN, and sodA genes, respectively. The optimized multiplex PCR demonstrated 100% analytical specificity, with a detection limit of 104 copies/μL for E. rhusiopathiae and S. hyicus, and 106 copies/μL for S. suis. The assay was further validated by comparison with previously established uniplex PCR assays and tested on field-collected pig blood samples to assess real-world applicability. The developed assay is rapid, specific and cost-effective and can serve as a handy diagnostic tool for testing suspect samples for these neglected bacterial pathogens of pigs.https://www.frontiersin.org/articles/10.3389/fvets.2025.1605316/fullmultiplex PCRErysipelothrix rhusiopathiaeStreptococcus suisStaphylococcus hyicuspigs |
| spellingShingle | A. Arun Prince Milton Sabia Khan Kandhan Srinivas Torik Basar Zakir Hussain G. Bhuvana Priya M. Saminathan L. H. Puii Sandeep Ghatak Samir Das Novel multiplex PCR approach for the detection of Erysipelothrix rhusiopathiae, Streptococcus suis, and Staphylococcus hyicus in swine Frontiers in Veterinary Science multiplex PCR Erysipelothrix rhusiopathiae Streptococcus suis Staphylococcus hyicus pigs |
| title | Novel multiplex PCR approach for the detection of Erysipelothrix rhusiopathiae, Streptococcus suis, and Staphylococcus hyicus in swine |
| title_full | Novel multiplex PCR approach for the detection of Erysipelothrix rhusiopathiae, Streptococcus suis, and Staphylococcus hyicus in swine |
| title_fullStr | Novel multiplex PCR approach for the detection of Erysipelothrix rhusiopathiae, Streptococcus suis, and Staphylococcus hyicus in swine |
| title_full_unstemmed | Novel multiplex PCR approach for the detection of Erysipelothrix rhusiopathiae, Streptococcus suis, and Staphylococcus hyicus in swine |
| title_short | Novel multiplex PCR approach for the detection of Erysipelothrix rhusiopathiae, Streptococcus suis, and Staphylococcus hyicus in swine |
| title_sort | novel multiplex pcr approach for the detection of erysipelothrix rhusiopathiae streptococcus suis and staphylococcus hyicus in swine |
| topic | multiplex PCR Erysipelothrix rhusiopathiae Streptococcus suis Staphylococcus hyicus pigs |
| url | https://www.frontiersin.org/articles/10.3389/fvets.2025.1605316/full |
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