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Immunomagnetic nanoparticle based quantitative PCR for rapid detection of Salmonella
Authors:Padmavathy Bakthavathsalam  Vinoth Kumar Rajendran  Uttara Saran  Suvro Chatterjee  Baquir Mohammed Jaffar Ali
Affiliation:1. AU-KBC Research Centre, M.I.T Campus of Anna University, Chennai, 600044, India
2. Centre for Green Energy Technology, Pondicherry University, Puducherry, 605014, India
Abstract:We have developed a rapid and sensitive method for immunomagnetic separation (IMS) of Salmonella along with their real time detection via PCR. Silica-coated magnetic nanoparticles were functionalized with carboxy groups to which anti-Salmonella antibody raised against heat-inactivated whole cells of Salmonella were covalently attached. The immuno-captured target cells were detected in beverages like milk and lemon juice by multiplex PCR and real time PCR with a detection limit of 104 cfu.mL?1 and 103 cfu.mL?1, respectively. We demonstrate that IMS can be used for selective concentration of target bacteria from beverages for subsequent use in PCR detection. PCR also enables differentiation of Salmonella typhi and Salmonella paratyphi A using a set of four specific primers. In addition, IMS—PCR can be used as a screening tool in the food and beverage industry for the detection of Salmonella within 3–4 h which compares favorably to the time of several days that is needed in case of conventional detection based on culture and biochemical methods.
The method uses silica coated magnetic nanoparticles immobilized with anti-Salmonella antibody for immunomagnetic separation of Salmonella from beverages followed by detection by multiplex PCR (mPCR) and real time PCR (qPCR). This methodology contributes to rapid screening and accurate detection of Salmonella contaminations in beverages.
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