Development of an Immunoassay for Detection of Staphylococcal Enterotoxin-Like J, A Non-Characterized Toxin
Staphylococcal enterotoxins (SEs) are the cause of staphylococcal food poisoning (SFP) outbreaks. Recently, many new types of SEs and SE-like toxins have been reported, but it has not been proved whether these new toxins cause food poisoning. To develop an immunoassay for detection of SE-like J (SEl...
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| Format: | Article |
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MDPI AG
2018-11-01
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| Series: | Toxins |
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| Online Access: | https://www.mdpi.com/2072-6651/10/11/458 |
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| author | Hisaya K. Ono Nobuaki Hachiya Yasunori Suzuki Ikunori Naito Shouhei Hirose Krisana Asano Katsuhiko Omoe Akio Nakane Dong-Liang Hu |
| author_facet | Hisaya K. Ono Nobuaki Hachiya Yasunori Suzuki Ikunori Naito Shouhei Hirose Krisana Asano Katsuhiko Omoe Akio Nakane Dong-Liang Hu |
| author_sort | Hisaya K. Ono |
| collection | DOAJ |
| description | Staphylococcal enterotoxins (SEs) are the cause of staphylococcal food poisoning (SFP) outbreaks. Recently, many new types of SEs and SE-like toxins have been reported, but it has not been proved whether these new toxins cause food poisoning. To develop an immunoassay for detection of SE-like J (SElJ), a non-characterized toxin in SFP, a mutant SElJ with C-terminus deletion (SElJ∆C) was expressed and purified in an <i>E. coli</i> expression system. Anti-SElJ antibody was produced in rabbits immunized with the SElJ∆C. Western blotting and sandwich enzyme-linked immunosorbent assay (ELISA) detection systems were established and showed that the antibody specifically recognizes SElJ without cross reaction to other SEs tested. The limit of detection for the sandwich ELISA was 0.078 ng/mL, showing high sensitivity. SElJ production in <i>S. aureus</i> was detected by using the sandwich ELISA and showed that <i>selj</i>-horboring isolates produced a large amount of SElJ in the culture supernatants, especially in that of the strain isolated from a food poisoning outbreak in Japan. These results demonstrate that the immunoassay for detection of SElJ is specific and sensitive and is useful for determining the native SElJ production in <i>S. aureus</i> isolated from food poisoning cases. |
| first_indexed | 2024-04-13T07:59:09Z |
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| institution | Directory Open Access Journal |
| issn | 2072-6651 |
| language | English |
| last_indexed | 2024-04-13T07:59:09Z |
| publishDate | 2018-11-01 |
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| record_format | Article |
| series | Toxins |
| spelling | doaj.art-77d3e8632d0b4eaba90326b6742126d52022-12-22T02:55:20ZengMDPI AGToxins2072-66512018-11-01101145810.3390/toxins10110458toxins10110458Development of an Immunoassay for Detection of Staphylococcal Enterotoxin-Like J, A Non-Characterized ToxinHisaya K. Ono0Nobuaki Hachiya1Yasunori Suzuki2Ikunori Naito3Shouhei Hirose4Krisana Asano5Katsuhiko Omoe6Akio Nakane7Dong-Liang Hu8Department of Zoonoses, Kitasato University School of Veterinary Medicine, 35-1 Higashi-23-ban-cho, Towada, Aomori 034-8628, JapanDepartment of Zoonoses, Kitasato University School of Veterinary Medicine, 35-1 Higashi-23-ban-cho, Towada, Aomori 034-8628, JapanDepartment of Microbiology, Tokyo Metropolitan Institute of Public Health, Shinjuku-ku, Tokyo 169-0073, JapanDepartment of Veterinary Medicine, Faculty of Agriculture, Iwate University, Ueda 3-18-8, Morioka, Iwate 020-8550, JapanDepartment of Microbiology and Immunology, Hirosaki University Graduate School of Medicine, 5 Zaifu-cho, Hirosaki 036-8562, JapanDepartment of Microbiology and Immunology, Hirosaki University Graduate School of Medicine, 5 Zaifu-cho, Hirosaki 036-8562, JapanDepartment of Veterinary Medicine, Faculty of Agriculture, Iwate University, Ueda 3-18-8, Morioka, Iwate 020-8550, JapanDepartment of Microbiology and Immunology, Hirosaki University Graduate School of Medicine, 5 Zaifu-cho, Hirosaki 036-8562, JapanDepartment of Zoonoses, Kitasato University School of Veterinary Medicine, 35-1 Higashi-23-ban-cho, Towada, Aomori 034-8628, JapanStaphylococcal enterotoxins (SEs) are the cause of staphylococcal food poisoning (SFP) outbreaks. Recently, many new types of SEs and SE-like toxins have been reported, but it has not been proved whether these new toxins cause food poisoning. To develop an immunoassay for detection of SE-like J (SElJ), a non-characterized toxin in SFP, a mutant SElJ with C-terminus deletion (SElJ∆C) was expressed and purified in an <i>E. coli</i> expression system. Anti-SElJ antibody was produced in rabbits immunized with the SElJ∆C. Western blotting and sandwich enzyme-linked immunosorbent assay (ELISA) detection systems were established and showed that the antibody specifically recognizes SElJ without cross reaction to other SEs tested. The limit of detection for the sandwich ELISA was 0.078 ng/mL, showing high sensitivity. SElJ production in <i>S. aureus</i> was detected by using the sandwich ELISA and showed that <i>selj</i>-horboring isolates produced a large amount of SElJ in the culture supernatants, especially in that of the strain isolated from a food poisoning outbreak in Japan. These results demonstrate that the immunoassay for detection of SElJ is specific and sensitive and is useful for determining the native SElJ production in <i>S. aureus</i> isolated from food poisoning cases.https://www.mdpi.com/2072-6651/10/11/458staphylococcal enterotoxinimmunoassayELISAfood poisoningsuperantigenic activity |
| spellingShingle | Hisaya K. Ono Nobuaki Hachiya Yasunori Suzuki Ikunori Naito Shouhei Hirose Krisana Asano Katsuhiko Omoe Akio Nakane Dong-Liang Hu Development of an Immunoassay for Detection of Staphylococcal Enterotoxin-Like J, A Non-Characterized Toxin Toxins staphylococcal enterotoxin immunoassay ELISA food poisoning superantigenic activity |
| title | Development of an Immunoassay for Detection of Staphylococcal Enterotoxin-Like J, A Non-Characterized Toxin |
| title_full | Development of an Immunoassay for Detection of Staphylococcal Enterotoxin-Like J, A Non-Characterized Toxin |
| title_fullStr | Development of an Immunoassay for Detection of Staphylococcal Enterotoxin-Like J, A Non-Characterized Toxin |
| title_full_unstemmed | Development of an Immunoassay for Detection of Staphylococcal Enterotoxin-Like J, A Non-Characterized Toxin |
| title_short | Development of an Immunoassay for Detection of Staphylococcal Enterotoxin-Like J, A Non-Characterized Toxin |
| title_sort | development of an immunoassay for detection of staphylococcal enterotoxin like j a non characterized toxin |
| topic | staphylococcal enterotoxin immunoassay ELISA food poisoning superantigenic activity |
| url | https://www.mdpi.com/2072-6651/10/11/458 |
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