Highly Sensitive Suspension Immunoassay for Multiplex Detection, Differentiation, and Quantification of Eight <i>Staphylococcus aureus</i> Enterotoxins (SEA to SEI)
Staphylococcal enterotoxins (SEs) are major contributors to foodborne intoxications. Reliable detection methods for SEs are essential to maintain food safety and protect public health. Since the heat-stable toxins also exert their toxic effect in the absence of the bacterium, reliance on DNA detecti...
Saved in:
| Main Authors: | , , , , , , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
MDPI AG
2025-05-01
|
| Series: | Toxins |
| Subjects: | |
| Online Access: | https://www.mdpi.com/2072-6651/17/6/265 |
| Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
| _version_ | 1849433885862002688 |
|---|---|
| author | Paulin Dettmann Martin Skiba Daniel Stern Jasmin Weisemann Hans Werner Mages Nadja Krez Martin B. Dorner Sara Schaarschmidt Marc A. Avondet Marcus Fulde Andreas Rummel Birgit Strommenger Sven Maurischat Brigitte G. Dorner |
| author_facet | Paulin Dettmann Martin Skiba Daniel Stern Jasmin Weisemann Hans Werner Mages Nadja Krez Martin B. Dorner Sara Schaarschmidt Marc A. Avondet Marcus Fulde Andreas Rummel Birgit Strommenger Sven Maurischat Brigitte G. Dorner |
| author_sort | Paulin Dettmann |
| collection | DOAJ |
| description | Staphylococcal enterotoxins (SEs) are major contributors to foodborne intoxications. Reliable detection methods for SEs are essential to maintain food safety and protect public health. Since the heat-stable toxins also exert their toxic effect in the absence of the bacterium, reliance on DNA detection alone can be misleading: it does not allow for determining which specific toxins encoded by a given strain are produced and epidemiologically linked with a given outbreak. Commercially available diagnostic assays for SE detection are so far limited in sensitivity and specificity as well as in the range of targeted toxins (SEA–SEE), thus non-targeted SEs linked to foodborne illness remain undetected at the protein level. This study aimed to develop a highly sensitive and specific multiplex suspension immunoassay (SIA) for SEA to SEI. To this end, high-affinity monoclonal antibodies (mAbs) for the specific detection of the individual SEs were generated. When implemented in sandwich ELISAs and multiplex SIA, these mAbs demonstrated exceptional sensitivity with detection limits in the low picogram per millilitre range. When applied for the analysis of SE production in liquid cultures of a panel of 145 whole-genome sequenced strains of <i>Staphylococcus</i> spp. and <i>Enterococcus faecalis</i>, the novel multiplex SIA detected and differentiated the eight SEs with assay accuracies of 86.9–100%. Notably, the multiplex SIA covered one to four sequence variants for each of the individual SEs. Validation confirmed high recovery rates and reliable performance in three representative complex food matrices. The implementation of the novel mAbs in a multiplex SIA enabled, for the first time, simultaneous detection, differentiation, and quantification of multiple SEs from minimal sample volumes using Luminex<sup>®</sup> technology. As a result, the multiplex SIA will help strengthen food safety protocols and public health response capabilities. |
| format | Article |
| id | doaj-art-0a32993ed73d4d7383e3682d751cd557 |
| institution | Kabale University |
| issn | 2072-6651 |
| language | English |
| publishDate | 2025-05-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Toxins |
| spelling | doaj-art-0a32993ed73d4d7383e3682d751cd5572025-08-20T03:26:52ZengMDPI AGToxins2072-66512025-05-0117626510.3390/toxins17060265Highly Sensitive Suspension Immunoassay for Multiplex Detection, Differentiation, and Quantification of Eight <i>Staphylococcus aureus</i> Enterotoxins (SEA to SEI)Paulin Dettmann0Martin Skiba1Daniel Stern2Jasmin Weisemann3Hans Werner Mages4Nadja Krez5Martin B. Dorner6Sara Schaarschmidt7Marc A. Avondet8Marcus Fulde9Andreas Rummel10Birgit Strommenger11Sven Maurischat12Brigitte G. Dorner13Centre for Biological Threats and Special Pathogens, Biological Toxins (ZBS3), Robert Koch Institute, 13353 Berlin, GermanyCentre for Biological Threats and Special Pathogens, Biological Toxins (ZBS3), Robert Koch Institute, 13353 Berlin, GermanyCentre for Biological Threats and Special Pathogens, Biological Toxins (ZBS3), Robert Koch Institute, 13353 Berlin, GermanyInstitute for Toxicology, Hannover Medical School, 30625 Hannover, GermanyCentre for Biological Threats and Special Pathogens, Biological Toxins (ZBS3), Robert Koch Institute, 13353 Berlin, GermanyInstitute for Toxicology, Hannover Medical School, 30625 Hannover, GermanyCentre for Biological Threats and Special Pathogens, Biological Toxins (ZBS3), Robert Koch Institute, 13353 Berlin, GermanyDepartment Biological Safety, German Federal Institute for Risk Assessment, 12277 Berlin, GermanyFederal Office for Civil Protection, Spiez Laboratory, 3700 Spiez, SwitzerlandCentre of Infection Medicine, Institute of Microbiology and Epizootics, Freie Universität Berlin, 14163 Berlin, GermanyInstitute for Toxicology, Hannover Medical School, 30625 Hannover, GermanyDepartment of Infectious Diseases, National Reference Centre (NRC) for Staphylococci and Enterococci, Division of Nosocomial Pathogens and Antibiotic Resistances, Robert Koch Institute, 38855 Wernigerode, GermanyDepartment Biological Safety, German Federal Institute for Risk Assessment, 12277 Berlin, GermanyCentre for Biological Threats and Special Pathogens, Biological Toxins (ZBS3), Robert Koch Institute, 13353 Berlin, GermanyStaphylococcal enterotoxins (SEs) are major contributors to foodborne intoxications. Reliable detection methods for SEs are essential to maintain food safety and protect public health. Since the heat-stable toxins also exert their toxic effect in the absence of the bacterium, reliance on DNA detection alone can be misleading: it does not allow for determining which specific toxins encoded by a given strain are produced and epidemiologically linked with a given outbreak. Commercially available diagnostic assays for SE detection are so far limited in sensitivity and specificity as well as in the range of targeted toxins (SEA–SEE), thus non-targeted SEs linked to foodborne illness remain undetected at the protein level. This study aimed to develop a highly sensitive and specific multiplex suspension immunoassay (SIA) for SEA to SEI. To this end, high-affinity monoclonal antibodies (mAbs) for the specific detection of the individual SEs were generated. When implemented in sandwich ELISAs and multiplex SIA, these mAbs demonstrated exceptional sensitivity with detection limits in the low picogram per millilitre range. When applied for the analysis of SE production in liquid cultures of a panel of 145 whole-genome sequenced strains of <i>Staphylococcus</i> spp. and <i>Enterococcus faecalis</i>, the novel multiplex SIA detected and differentiated the eight SEs with assay accuracies of 86.9–100%. Notably, the multiplex SIA covered one to four sequence variants for each of the individual SEs. Validation confirmed high recovery rates and reliable performance in three representative complex food matrices. The implementation of the novel mAbs in a multiplex SIA enabled, for the first time, simultaneous detection, differentiation, and quantification of multiple SEs from minimal sample volumes using Luminex<sup>®</sup> technology. As a result, the multiplex SIA will help strengthen food safety protocols and public health response capabilities.https://www.mdpi.com/2072-6651/17/6/265staphylococcal enterotoxinfood poisoningfood safetymonoclonal antibodiesmultiplexsuspension immunoassay |
| spellingShingle | Paulin Dettmann Martin Skiba Daniel Stern Jasmin Weisemann Hans Werner Mages Nadja Krez Martin B. Dorner Sara Schaarschmidt Marc A. Avondet Marcus Fulde Andreas Rummel Birgit Strommenger Sven Maurischat Brigitte G. Dorner Highly Sensitive Suspension Immunoassay for Multiplex Detection, Differentiation, and Quantification of Eight <i>Staphylococcus aureus</i> Enterotoxins (SEA to SEI) Toxins staphylococcal enterotoxin food poisoning food safety monoclonal antibodies multiplex suspension immunoassay |
| title | Highly Sensitive Suspension Immunoassay for Multiplex Detection, Differentiation, and Quantification of Eight <i>Staphylococcus aureus</i> Enterotoxins (SEA to SEI) |
| title_full | Highly Sensitive Suspension Immunoassay for Multiplex Detection, Differentiation, and Quantification of Eight <i>Staphylococcus aureus</i> Enterotoxins (SEA to SEI) |
| title_fullStr | Highly Sensitive Suspension Immunoassay for Multiplex Detection, Differentiation, and Quantification of Eight <i>Staphylococcus aureus</i> Enterotoxins (SEA to SEI) |
| title_full_unstemmed | Highly Sensitive Suspension Immunoassay for Multiplex Detection, Differentiation, and Quantification of Eight <i>Staphylococcus aureus</i> Enterotoxins (SEA to SEI) |
| title_short | Highly Sensitive Suspension Immunoassay for Multiplex Detection, Differentiation, and Quantification of Eight <i>Staphylococcus aureus</i> Enterotoxins (SEA to SEI) |
| title_sort | highly sensitive suspension immunoassay for multiplex detection differentiation and quantification of eight i staphylococcus aureus i enterotoxins sea to sei |
| topic | staphylococcal enterotoxin food poisoning food safety monoclonal antibodies multiplex suspension immunoassay |
| url | https://www.mdpi.com/2072-6651/17/6/265 |
| work_keys_str_mv | AT paulindettmann highlysensitivesuspensionimmunoassayformultiplexdetectiondifferentiationandquantificationofeightistaphylococcusaureusienterotoxinsseatosei AT martinskiba highlysensitivesuspensionimmunoassayformultiplexdetectiondifferentiationandquantificationofeightistaphylococcusaureusienterotoxinsseatosei AT danielstern highlysensitivesuspensionimmunoassayformultiplexdetectiondifferentiationandquantificationofeightistaphylococcusaureusienterotoxinsseatosei AT jasminweisemann highlysensitivesuspensionimmunoassayformultiplexdetectiondifferentiationandquantificationofeightistaphylococcusaureusienterotoxinsseatosei AT hanswernermages highlysensitivesuspensionimmunoassayformultiplexdetectiondifferentiationandquantificationofeightistaphylococcusaureusienterotoxinsseatosei AT nadjakrez highlysensitivesuspensionimmunoassayformultiplexdetectiondifferentiationandquantificationofeightistaphylococcusaureusienterotoxinsseatosei AT martinbdorner highlysensitivesuspensionimmunoassayformultiplexdetectiondifferentiationandquantificationofeightistaphylococcusaureusienterotoxinsseatosei AT saraschaarschmidt highlysensitivesuspensionimmunoassayformultiplexdetectiondifferentiationandquantificationofeightistaphylococcusaureusienterotoxinsseatosei AT marcaavondet highlysensitivesuspensionimmunoassayformultiplexdetectiondifferentiationandquantificationofeightistaphylococcusaureusienterotoxinsseatosei AT marcusfulde highlysensitivesuspensionimmunoassayformultiplexdetectiondifferentiationandquantificationofeightistaphylococcusaureusienterotoxinsseatosei AT andreasrummel highlysensitivesuspensionimmunoassayformultiplexdetectiondifferentiationandquantificationofeightistaphylococcusaureusienterotoxinsseatosei AT birgitstrommenger highlysensitivesuspensionimmunoassayformultiplexdetectiondifferentiationandquantificationofeightistaphylococcusaureusienterotoxinsseatosei AT svenmaurischat highlysensitivesuspensionimmunoassayformultiplexdetectiondifferentiationandquantificationofeightistaphylococcusaureusienterotoxinsseatosei AT brigittegdorner highlysensitivesuspensionimmunoassayformultiplexdetectiondifferentiationandquantificationofeightistaphylococcusaureusienterotoxinsseatosei |