Polydiacetylene for the Detection of alpha-Hemolysin in Milk toward the Diagnosis of Bovine Mastitis

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dc.authorid0000-0003-1858-0691en_US
dc.contributor.authorWeston, Max
dc.contributor.authorÇiftçi, Mustafa
dc.contributor.authorKuchel, Rhiannon P.
dc.contributor.authorBoyer, Cyrille
dc.contributor.authorChandrawati, Rona
dc.date.accessioned2021-03-20T20:09:16Z
dc.date.available2021-03-20T20:09:16Z
dc.date.issued2020
dc.departmentBTÜ, Mühendislik ve Doğa Bilimleri Fakültesi, Kimya Bölümüen_US
dc.description.abstractEarly diagnosis of mastitis is essential to reduce the expenses it imparts on the dairy industry. Existing techniques fail to provide rapid and pathogen-specific information to direct the prescription of antibiotics. Polydiacetylene (PDA)-based colorimetric sensors have emerged as promising bacterial screening tools that show potential for application in the diagnosis of bacterial infections. However, there are challenges in operating PDA sensors in complex samples, as identified in our recent work. To overcome this challenge, herein, we design and synthesize a custom diacetylene monomer via a copper-catalyzed cross-coupling reaction to yield 6,8-tricosadiynoic acid. Photopolymerization of 6,8-tricosadiynoic acid monomers yields blue-phase PDA with superior chromatic stability to milk constituents and reduced chromatic response to temperature fluctuations, which diminish the likelihood of false positives. We then form functionalized PDA vesicles by insertion of 1,2-dimyristoyl-sn-glycero-3-phosphocholine phospholipid and cholesterol in the PDA membrane. This allows the detection of alpha-hemolysin, a hemolytic toxin excreted by Staphylococcus aureus, a predominant causative pathogen of bovine mastitis. We demonstrate the detection of alpha-hemolysin in phosphate-buffered saline (limit of detection (LOD) = 3.62 mu g/mL) and milk samples (LOD = 6.62 mu g/mL) by a blue to red color change visible to the naked eye. The color change is studied by absorption spectroscopy and digital colorimetric analysis of photographs. We attribute the color change to pore formation in the PDA membrane and lysis of the vesicles due to the action of the S. aureus alpha-hemolysin. This is evidenced by vesicle membrane destruction in transmission electron microscopy micrographs of PDA vesicles before and after incubation with the toxin. The specificity of the sensor is demonstrated by discrimination between alpha-hemolysin and other toxins and biomarkers for mastitis. This is the first custom-designed PDA for point-of-care application in bovine milk samples and the first documented detection of alpha-hemolysin using a PDA-based sensor.en_US
dc.description.sponsorshipAustralian Government Research Training Program ScholarshipAustralian GovernmentDepartment of Industry, Innovation and Science; National Health and Medical Research Council Emerging Leadership Investigator Grant (NHMRC)National Health and Medical Research Council of Australia [EL2 APP1173428]; Australian Research Council Discovery Early Career Researcher Award (ARC DECRA)Australian Research Council [DE170100068]; UNSW Scientia Fellowshipen_US
dc.description.sponsorshipM.W. acknowledges support from the Australian Government Research Training Program Scholarship. R.C. acknowledges support from the National Health and Medical Research Council Emerging Leadership Investigator Grant (NHMRC EL2 APP1173428), the Australian Research Council Discovery Early Career Researcher Award (ARC DECRA DE170100068), and the UNSW Scientia Fellowship. The authors thank Yuan Gao for assistance with FTIR characterization. This research used the facilities at the Mark Wainwright Analytical Centre Electron Microscope Unit at UNSW.en_US
dc.identifier.doi10.1021/acsapm.0c00968en_US
dc.identifier.endpage5248en_US
dc.identifier.issn2637-6105
dc.identifier.issue11en_US
dc.identifier.scopusqualityQ1en_US
dc.identifier.startpage5238en_US
dc.identifier.urihttp://doi.org/10.1021/acsapm.0c00968
dc.identifier.urihttps://hdl.handle.net/20.500.12885/340
dc.identifier.volume2en_US
dc.identifier.wosWOS:000592755800093en_US
dc.identifier.wosqualityQ1en_US
dc.indekslendigikaynakWeb of Scienceen_US
dc.indekslendigikaynakScopusen_US
dc.institutionauthorÇiftçi, Mustafa
dc.language.isoenen_US
dc.publisherAmer Chemical Socen_US
dc.relation.ispartofAcs Applied Polymer Materialsen_US
dc.relation.publicationcategoryMakale - Uluslararası Hakemli Dergi - Kurum Öğretim Elemanıen_US
dc.rightsinfo:eu-repo/semantics/closedAccessen_US
dc.subjectpolydiacetyleneen_US
dc.subjectsensoren_US
dc.subjecttoxinen_US
dc.subjectalpha-hemolysinen_US
dc.subjectmastitisen_US
dc.subjectcolorimetricen_US
dc.titlePolydiacetylene for the Detection of alpha-Hemolysin in Milk toward the Diagnosis of Bovine Mastitisen_US
dc.typeArticleen_US

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