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A dual-mode biosensor was constructed using two phage probes and portable bioluminescence/fluorescence detectors. A magnetic phage capture probe and a fluorescently labeled signal tag form magnetic sandwich complexes, whose fluorescence reports total S.T after magnetic separation. Lysozyme then decomposes captured live S.T to release ATP and generate bioluminescence for live counts; dead levels follow total minus live. Detection limits were 55 CFU/mL (total) and 9 CFU/mL (live) within 20 minutes, demonstrated in milk and adaptable to other targets by changing phages.",{"@graph":69,"@context":123},[70,84,106],{"@type":71,"itemListElement":72},"BreadcrumbList",[73,77,79,82],{"item":74,"name":75,"@type":76,"position":8},"https://docshare.wps.com","Home","ListItem",{"item":78,"name":9,"@type":76,"position":14},"https://docshare.wps.com/document/",{"item":80,"name":35,"@type":76,"position":81},"https://docshare.wps.com/document/healthcare/",3,{"item":83,"name":65,"@type":76,"position":19},"https://docshare.wps.com/document/dual-mode-biosensor-for-simultaneous-and-rapid-detection-of-live-and-whole-salmonella-typhimurium-based-on-bioluminescence-and-fluorescence-detection/156500/",{"url":83,"name":65,"@type":85,"image":86,"author":91,"headline":65,"publisher":94,"fileFormat":97,"inLanguage":63,"description":67,"dateModified":98,"datePublished":99,"encodingFormat":97,"isAccessibleForFree":100,"interactionStatistic":101},"DigitalDocument",{"url":87,"@type":88,"width":89,"height":90},"https://docshare.wps.com/thumbnails/dual-mode-biosensor-for-simultaneous-and-rapid-detection-of-live-and-whole-salmonella-typhimurium-based-on-bioluminescence-and-fluorescence-detection/156500.png","ImageObject",300,407,{"name":92,"@type":93},"Ophelia","Person",{"url":74,"name":95,"@type":96},"DocShare","Organization","application/pdf","2026-10-23","2026-08-28",true,{"@type":102,"interactionType":103,"userInteractionCount":105},"InteractionCounter",{"@type":104},"ViewAction",12,{"@type":107,"mainEntity":108},"FAQPage",[109,115,119],{"name":110,"@type":111,"acceptedAnswer":112},"Why is it important to distinguish live versus dead Salmonella typhimurium in food safety?","Question",{"text":113,"@type":114},"Live and dead S.T are both harmful but differ in pathological mechanism, dosage, and security. Accurate discrimination supports choosing appropriate response protocols in food safety cases.","Answer",{"name":116,"@type":111,"acceptedAnswer":117},"How does the dual-mode biosensor measure total and live S.T simultaneously?",{"text":118,"@type":114},"Magnetic capture forms a sandwich complex with phage probes; fluorescence under 365 nm laser reports total S.T. Lysozyme decomposes captured live bacteria to release ATP, generating bioluminescence proportional to the live S.T amount.",{"name":120,"@type":111,"acceptedAnswer":121},"What are the reported detection limits and how fast is the assay?",{"text":122,"@type":114},"The detection limit was 55 CFU/mL for total S.T and 9 CFU/mL for live bacteria, with results obtained within 20 minutes.","https://schema.org",{"og:url":83,"og:type":125,"og:title":65,"og:site_name":95,"og:description":67},"article",{"robots":127,"canonical":83},"index,follow",{"doc_id":129,"site_id":62},156500,1787961005,{"code":4,"msg":5,"data":132},{"doc_id":129,"user_id":133,"nickname":92,"user_avatar":134,"doc_module":4,"category_id":34,"category_name":35,"doc_title":65,"doc_description":67,"doc_content":135,"file_id":136,"file_url":137,"file_type":138,"file_size":139,"view_count":105,"is_deleted":4,"is_public":8,"is_downloadable":8,"audit_status":8,"page_count":140,"language":141,"language_code":63,"site_id":62,"html_lang":63,"table_of_contents":142,"faqs":143,"seo_title":144,"seo_description":67,"update_tm":130,"read_time":145},7971461741311,"https://ap-avatar.wpscdn.com/avatar/74000253aff267980c6?x-image-process=image/resize,m_fixed,w_180,h_180&k=1779345379180704826","biosensors  \nArticle  \nDual-Mode Biosensor for Simultaneous and Rapid Detection of Live and Whole Salmonella typhimurium Based on Bioluminescence and Fluorescence Detection  \nZhenli Xu 1, Bailu Liu 1, Dengfeng Li 2, Zhenzhong Yu 1 and Ning Gan 1, *  \nCitation: Xu, Z.; Liu, B.; Li, D.; Yu, Z.; Gan, N. Dual-Mode Biosensor for Simultaneous and Rapid Detection of Live and Whole Salmonella typhimurium Based on Bioluminescence and Fluorescence Detection. Biosensors 2023, 13, 401. [https://doi.org/](https://doi.org/)[ ](https://doi.org/)[10.3390/bios13030401](10.3390/bios13030401)  \nReceived: 12 February 2023  \nRevised: 10 March 2023  \nAccepted: 18 March 2023  \nPublished: 19 March 2023  \nCopyright: © 2023 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license ([https://](https://)[ ](https://)[creativecommons.org/licenses/by/](creativecommons.org/licenses/by/)[ ](creativecommons.org/licenses/by/)[4.0/](4.0/)) .  \n1 Faculty of Material Science and Chemical Engineering, Ningbo University, Ningbo 315211, China  \n2 Faculty of Marine, Ningbo University, Ningbo 315211, China  \n* Correspondence: [ganning@nbu.edu.cn](ganning@nbu.edu.cn)  \nAbstract: Both live and dead Salmonella typhimurium (S.T) are harmful to human health, but there are differences in pathological mechanism, dosage, and security. It is crucial to develop a rapid and simultaneous assay to distinguish and quantify live and dead S.T in foods. Herein, one dual-mode biosensor for simultaneous detection of live and dead S.T was fabricated based on two phage probes, using portable bioluminescence and fluorescent meter as detectors, respectively. Firstly, a magnetic phage capture probe (M-P1) and a phage signal tag (P2-S) labeled with SYTO 13 fluorescent dye were prepared, respectively. Both M-P1 and P2-S can specifically conjugate with S.T to form a magnetic sandwich complex. After magnetic separation, the isolated complex can emit a fluorescent signal under an excited 365 nm laser, which can reflect the total amount of S.T. Afterwards, the lysozyme was added to decompose the captured live S.T, which can release ATP and produce a bioluminescent signal corresponding to the live S.T amount. The dead S.T concentration can be deduced by the difference between total and live examples. The detection limit of 55 CFU/mL for total S.T and 9 CFU/mL for live ones was within 20 min. The assay was successfully employed in milk samples and prospectively for on-site screening of other dead and live bacteria, while changing the phages for the targets.  \nKeywords: dual-mode biosensor; dead and live bacteria; Salmonella typhimurium; magnetic phages; portable ATP bioluminescence; fluorescence meter  \n1. Introduction  \nOutbreaks of foodborne pathogens in foods can lead to serious infectious diseases, which have been a great public health threat worldwide [1] . Especially, Salmonella, as agram-negative bacterium, ranks first among all types of bacterial food poisoning [2] . It was estimated that about 93 .8 million Salmonella infections and 155,000 related deaths occur each year worldwide [3] . Among these, Salmonella typhimurium (S.T) is one of the most common serotypes [4] . According to [5], both live and dead S.T are harmful to human health, but there are differences in pathological mechanism, dosage, and security. Live S.T can cause large-scale contamination of foods within a short time, attributed to fast reproduction speed and strong environmental resistance [6,7] . When S.T dies or lyses, it can release endotoxins to cause bacterial virulence and pose a serious health risk to the host [8] . Therefore, effective detection and discrimination of live and dead S.T is essential for selecting an appropriate response protocol in food safety cases. At present, several methods have been developed to detect pathogenic bacteria, such as the plate counting method (the gold standard), ","cbCaiv4pL1Wlgv3q","https://ap.wps.com/l/cbCaiv4pL1Wlgv3q","pdf",5409029,15,"English","# Introduction\n## Foodborne risks and need for live/dead discrimination\n## Limitations of conventional detection methods\n## Rationale for ATP bioluminescence and phage-based selectivity\n# Materials and Methods\n## Dual-mode biosensor concept\n## Magnetic phage capture and fluorescent signaling\n## Lysozyme-assisted ATP release for live detection\n# Results and Discussion\n## Detection performance and limits\n## Application to milk samples and on-site screening","[{\"question\":\"Why is it important to distinguish live versus dead Salmonella typhimurium in food safety?\",\"answer\":\"Live and dead S.T are both harmful but differ in pathological mechanism, dosage, and security. Accurate discrimination supports choosing appropriate response protocols in food safety cases.\"},{\"question\":\"How does the dual-mode biosensor measure total and live S.T simultaneously?\",\"answer\":\"Magnetic capture forms a sandwich complex with phage probes; fluorescence under 365 nm laser reports total S.T. Lysozyme decomposes captured live bacteria to release ATP, generating bioluminescence proportional to the live S.T amount.\"},{\"question\":\"What are the reported detection limits and how fast is the assay?\",\"answer\":\"The detection limit was 55 CFU/mL for total S.T and 9 CFU/mL for live bacteria, with results obtained within 20 minutes.\"}]","Dual-Mode Biosensor for Simultaneous and Rapid Detection of Live and Whole Salmonella typhimurium Based on Bioluminescence and Fluorescence Detection | PDF",38]