Bifunctionalized nanobioprobe based rapid color-shift assay for typhoid targeting Vi capsular polysaccharide
DIR@IMTECH: CSIR-Institute of Microbial Technology
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Title |
Bifunctionalized nanobioprobe based rapid color-shift assay for typhoid targeting Vi capsular polysaccharide
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Creator |
Choudhary, Megha
Bisht, Bhawana Saini, Jai Kumar Bharti, . Singh, Pargat Bhardwaj, Priya Dilawari, Rahul Pinnaka, Anil Kumar Ray, Pallab Gupta, Madhu Sethi, Sunil Suri, C Raman Raje, Manoj Bhalla, Vijayender |
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Subject |
QR Microbiology
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Description |
Typhoid fever is an acute illness caused by Salmonella Typhi and the current diagnostic gap leads to inaccurate, over-diagnosis of typhoid leading to excessive use of antibiotics. Herein, to address the challenges we describe a new rapid color-shift assay based on a novel bifunctional nanobioprobe (Vi-AgNP probe) that is functionalized with specific biomarker Vi polysaccharide and also has the co-presence of Ag as urease inhibitor. The immunoreactions between the Vi with specific antibodies (Abs) present in typhoid patient sample forms a shielding barrier over Vi-AgNP probe rendering the urease to be active, generating colored output. Vi polysaccharide coating on the AgNP was visualized using HRTEM. TEM was performed to get insight into shielding barrier formation by the Abs. MST (microscale thermophoresis) data showed less binding Kd of 7.43 μM in presence of Abs whereas probe with urease showed efficient binding with Kd 437 nM. The assay was validated using 53 human sera samples and proven effective with 100% sensitivity. The assay showed relative standard deviation (RSD) of 4.3% estimated using rabbit anti-Vi Abs. The entire procedure could be completed within 15 min. Unlike lateral flow based assays, our assay does not require multiple combination of Abs for detection. The assay format was also found compatible in paper strip test that provides promising opportunities to develop low-cost on-spot assay for clinical diagnostics.
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Publisher |
Elsevier Science
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Date |
2023-05-15
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Type |
Article
PeerReviewed |
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Relation |
https://www.sciencedirect.com/science/article/pii/S0956566323001379?via%3Dihub
http://crdd.osdd.net/open/3079/ |
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Identifier |
Choudhary, Megha and Bisht, Bhawana and Saini, Jai Kumar and Bharti, . and Singh, Pargat and Bhardwaj, Priya and Dilawari, Rahul and Pinnaka, Anil Kumar and Ray, Pallab and Gupta, Madhu and Sethi, Sunil and Suri, C Raman and Raje, Manoj and Bhalla, Vijayender (2023) Bifunctionalized nanobioprobe based rapid color-shift assay for typhoid targeting Vi capsular polysaccharide. Biosensors & bioelectronics, 228.
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