Novel helix loop-mediated isothermal amplification (HAMP) assay for colorimetric detection of Staphylococcus aureus in milk
Staphylococcus aureus is an important and leading cause of foodborne diseases worldwide. Prompt detection and recall of contaminated foods are crucial to prevent untoward health consequences caused by S. aureus . Helix loop-mediated isothermal amplification (HAMP) is an exciting recent addition to t...
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| Veröffentlicht in: | World journal of microbiology & biotechnology 2024-01, Vol.40 (1), p.14-14, Article 14 |
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| container_title | World journal of microbiology & biotechnology |
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| creator | Milton, A. Arun Prince Prasad, M.C.B. Priya, G. Bhuvana Momin, K.M. Lyngdoh, Vanita Srinivas, K. Das, Samir Ghatak, Sandeep |
| description | Staphylococcus aureus
is an important and leading cause of foodborne diseases worldwide. Prompt detection and recall of contaminated foods are crucial to prevent untoward health consequences caused by
S. aureus
. Helix loop-mediated isothermal amplification (HAMP) is an exciting recent addition to the array of available isothermal-based nucleic acid amplification techniques. This study aimed to develop and evaluate a HAMP assay for detecting
S. aureus
in milk and milk products. The assay is completed in 75 minutes of isothermal temperature incubation (64 ˚C) and dye-based visual interpretation of results based on colour change. The specificity of the developed assay was ascertained using 27
S. aureus
and 17 non
S. aureus
bacterial strains. The analytical sensitivity of the developed HAMP assay was 9.7 fg/µL of pure
S. aureus
DNA. The detection limit of the HAMP assay in milk (86 CFU/mL) was 1000x greater than the routinely used endpoint PCR (86 × 10
3
CFU/mL). The practicality of applying the HAMP assay was also assessed by analysing milk and milk product samples (n = 95) obtained from different dairy farms and retail outlets. The developed test is a more rapid, sensitive, and user-friendly method for the high-throughput screening of
S. aureus
in food samples and may therefore be suitable for field laboratories. To our knowledge, this is the first study to develop and evaluate the HAMP platform for detecting
S. aureus. |
| doi_str_mv | 10.1007/s11274-023-03838-3 |
| format | Article |
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is an important and leading cause of foodborne diseases worldwide. Prompt detection and recall of contaminated foods are crucial to prevent untoward health consequences caused by
S. aureus
. Helix loop-mediated isothermal amplification (HAMP) is an exciting recent addition to the array of available isothermal-based nucleic acid amplification techniques. This study aimed to develop and evaluate a HAMP assay for detecting
S. aureus
in milk and milk products. The assay is completed in 75 minutes of isothermal temperature incubation (64 ˚C) and dye-based visual interpretation of results based on colour change. The specificity of the developed assay was ascertained using 27
S. aureus
and 17 non
S. aureus
bacterial strains. The analytical sensitivity of the developed HAMP assay was 9.7 fg/µL of pure
S. aureus
DNA. The detection limit of the HAMP assay in milk (86 CFU/mL) was 1000x greater than the routinely used endpoint PCR (86 × 10
3
CFU/mL). The practicality of applying the HAMP assay was also assessed by analysing milk and milk product samples (n = 95) obtained from different dairy farms and retail outlets. The developed test is a more rapid, sensitive, and user-friendly method for the high-throughput screening of
S. aureus
in food samples and may therefore be suitable for field laboratories. To our knowledge, this is the first study to develop and evaluate the HAMP platform for detecting
S. aureus.</description><identifier>ISSN: 0959-3993</identifier><identifier>EISSN: 1573-0972</identifier><identifier>DOI: 10.1007/s11274-023-03838-3</identifier><identifier>PMID: 37966568</identifier><language>eng</language><publisher>Dordrecht: Springer Netherlands</publisher><subject>Animals ; Applied Microbiology ; Assaying ; Biochemistry ; Biomedical and Life Sciences ; Biotechnology ; color ; Colorimetry ; Dairy farms ; detection limit ; DNA ; Environmental Engineering/Biotechnology ; Food contamination ; Foodborne diseases ; Gene amplification ; Hepcidins ; High-throughput screening ; Humans ; Life Sciences ; loop-mediated isothermal amplification ; Microbiology ; Milk ; Milk products ; Nucleic Acid Amplification Techniques ; Nucleic acids ; Sensitivity analysis ; Staphylococcal Infections ; Staphylococcus aureus ; Staphylococcus aureus - genetics ; temperature</subject><ispartof>World journal of microbiology & biotechnology, 2024-01, Vol.40 (1), p.14-14, Article 14</ispartof><rights>The Author(s), under exclusive licence to Springer Nature B.V. 2023. Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.</rights><rights>2023. The Author(s), under exclusive licence to Springer Nature B.V.</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><cites>FETCH-LOGICAL-c359t-6634003b6e2ef4091b8a229be081711249dce4212b5824bf3018aa9be5e3e20a3</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://link.springer.com/content/pdf/10.1007/s11274-023-03838-3$$EPDF$$P50$$Gspringer$$H</linktopdf><linktohtml>$$Uhttps://link.springer.com/10.1007/s11274-023-03838-3$$EHTML$$P50$$Gspringer$$H</linktohtml><link.rule.ids>314,776,780,27901,27902,41464,42533,51294</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/37966568$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Milton, A. Arun Prince</creatorcontrib><creatorcontrib>Prasad, M.C.B.</creatorcontrib><creatorcontrib>Priya, G. Bhuvana</creatorcontrib><creatorcontrib>Momin, K.M.</creatorcontrib><creatorcontrib>Lyngdoh, Vanita</creatorcontrib><creatorcontrib>Srinivas, K.</creatorcontrib><creatorcontrib>Das, Samir</creatorcontrib><creatorcontrib>Ghatak, Sandeep</creatorcontrib><title>Novel helix loop-mediated isothermal amplification (HAMP) assay for colorimetric detection of Staphylococcus aureus in milk</title><title>World journal of microbiology & biotechnology</title><addtitle>World J Microbiol Biotechnol</addtitle><addtitle>World J Microbiol Biotechnol</addtitle><description>Staphylococcus aureus
is an important and leading cause of foodborne diseases worldwide. Prompt detection and recall of contaminated foods are crucial to prevent untoward health consequences caused by
S. aureus
. Helix loop-mediated isothermal amplification (HAMP) is an exciting recent addition to the array of available isothermal-based nucleic acid amplification techniques. This study aimed to develop and evaluate a HAMP assay for detecting
S. aureus
in milk and milk products. The assay is completed in 75 minutes of isothermal temperature incubation (64 ˚C) and dye-based visual interpretation of results based on colour change. The specificity of the developed assay was ascertained using 27
S. aureus
and 17 non
S. aureus
bacterial strains. The analytical sensitivity of the developed HAMP assay was 9.7 fg/µL of pure
S. aureus
DNA. The detection limit of the HAMP assay in milk (86 CFU/mL) was 1000x greater than the routinely used endpoint PCR (86 × 10
3
CFU/mL). The practicality of applying the HAMP assay was also assessed by analysing milk and milk product samples (n = 95) obtained from different dairy farms and retail outlets. The developed test is a more rapid, sensitive, and user-friendly method for the high-throughput screening of
S. aureus
in food samples and may therefore be suitable for field laboratories. To our knowledge, this is the first study to develop and evaluate the HAMP platform for detecting
S. aureus.</description><subject>Animals</subject><subject>Applied Microbiology</subject><subject>Assaying</subject><subject>Biochemistry</subject><subject>Biomedical and Life Sciences</subject><subject>Biotechnology</subject><subject>color</subject><subject>Colorimetry</subject><subject>Dairy farms</subject><subject>detection limit</subject><subject>DNA</subject><subject>Environmental Engineering/Biotechnology</subject><subject>Food contamination</subject><subject>Foodborne diseases</subject><subject>Gene amplification</subject><subject>Hepcidins</subject><subject>High-throughput screening</subject><subject>Humans</subject><subject>Life Sciences</subject><subject>loop-mediated isothermal amplification</subject><subject>Microbiology</subject><subject>Milk</subject><subject>Milk products</subject><subject>Nucleic Acid Amplification Techniques</subject><subject>Nucleic acids</subject><subject>Sensitivity analysis</subject><subject>Staphylococcal Infections</subject><subject>Staphylococcus aureus</subject><subject>Staphylococcus aureus - genetics</subject><subject>temperature</subject><issn>0959-3993</issn><issn>1573-0972</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2024</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><sourceid>BENPR</sourceid><recordid>eNqFkcFu1DAQhi0EokvhBTggS1zaQ1rb4zjxsapKi1QoEnC2HO-EdXHiYCeIVV8ed7eAxAFOI2u--a2Zj5CXnJ1wxprTzLloZMUEVAxaaCt4RFa8bspTN-IxWTFd6wq0hgPyLOdbxsqYhqfkABqtVK3aFbl7H79joBsM_gcNMU7VgGtvZ1xTn-O8wTTYQO0wBd97Z2cfR3p0dfbuwzG1Odst7WOiLoaY_IBz8o6ucUa342JPP8522mxDdNG5JVO7JCzFj3Tw4etz8qS3IeOLh3pIPr-5-HR-VV3fXL49P7uuHNR6rpQCyRh0CgX2kmnetVYI3SFreVNOIPXaoRRcdHUrZNcD4621pV8joGAWDsnRPndK8duCeTaDzw5DsCPGJRvgNaiWNbX4LypazUBxJqGgr_9Cb-OSxrLIjuKKS6kKJfaUSzHnhL2ZyqVs2hrOzL1Fs7doikWzs2juo189RC9d0fF75Je2AsAeyKU1fsH05-9_xP4EiZGnkg</recordid><startdate>20240101</startdate><enddate>20240101</enddate><creator>Milton, A. 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Arun Prince</au><au>Prasad, M.C.B.</au><au>Priya, G. Bhuvana</au><au>Momin, K.M.</au><au>Lyngdoh, Vanita</au><au>Srinivas, K.</au><au>Das, Samir</au><au>Ghatak, Sandeep</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Novel helix loop-mediated isothermal amplification (HAMP) assay for colorimetric detection of Staphylococcus aureus in milk</atitle><jtitle>World journal of microbiology & biotechnology</jtitle><stitle>World J Microbiol Biotechnol</stitle><addtitle>World J Microbiol Biotechnol</addtitle><date>2024-01-01</date><risdate>2024</risdate><volume>40</volume><issue>1</issue><spage>14</spage><epage>14</epage><pages>14-14</pages><artnum>14</artnum><issn>0959-3993</issn><eissn>1573-0972</eissn><abstract>Staphylococcus aureus
is an important and leading cause of foodborne diseases worldwide. Prompt detection and recall of contaminated foods are crucial to prevent untoward health consequences caused by
S. aureus
. Helix loop-mediated isothermal amplification (HAMP) is an exciting recent addition to the array of available isothermal-based nucleic acid amplification techniques. This study aimed to develop and evaluate a HAMP assay for detecting
S. aureus
in milk and milk products. The assay is completed in 75 minutes of isothermal temperature incubation (64 ˚C) and dye-based visual interpretation of results based on colour change. The specificity of the developed assay was ascertained using 27
S. aureus
and 17 non
S. aureus
bacterial strains. The analytical sensitivity of the developed HAMP assay was 9.7 fg/µL of pure
S. aureus
DNA. The detection limit of the HAMP assay in milk (86 CFU/mL) was 1000x greater than the routinely used endpoint PCR (86 × 10
3
CFU/mL). The practicality of applying the HAMP assay was also assessed by analysing milk and milk product samples (n = 95) obtained from different dairy farms and retail outlets. The developed test is a more rapid, sensitive, and user-friendly method for the high-throughput screening of
S. aureus
in food samples and may therefore be suitable for field laboratories. To our knowledge, this is the first study to develop and evaluate the HAMP platform for detecting
S. aureus.</abstract><cop>Dordrecht</cop><pub>Springer Netherlands</pub><pmid>37966568</pmid><doi>10.1007/s11274-023-03838-3</doi><tpages>1</tpages></addata></record> |
| fulltext | fulltext |
| identifier | ISSN: 0959-3993 |
| ispartof | World journal of microbiology & biotechnology, 2024-01, Vol.40 (1), p.14-14, Article 14 |
| issn | 0959-3993 1573-0972 |
| language | eng |
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| source | MEDLINE; Springer Journals |
| subjects | Animals Applied Microbiology Assaying Biochemistry Biomedical and Life Sciences Biotechnology color Colorimetry Dairy farms detection limit DNA Environmental Engineering/Biotechnology Food contamination Foodborne diseases Gene amplification Hepcidins High-throughput screening Humans Life Sciences loop-mediated isothermal amplification Microbiology Milk Milk products Nucleic Acid Amplification Techniques Nucleic acids Sensitivity analysis Staphylococcal Infections Staphylococcus aureus Staphylococcus aureus - genetics temperature |
| title | Novel helix loop-mediated isothermal amplification (HAMP) assay for colorimetric detection of Staphylococcus aureus in milk |
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