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Microchimica Acta

, Volume 180, Issue 13–14, pp 1241–1248 | Cite as

Immunomagnetic nanoparticle based quantitative PCR for rapid detection of Salmonella

  • Padmavathy Bakthavathsalam
  • Vinoth Kumar Rajendran
  • Uttara Saran
  • Suvro Chatterjee
  • Baquir Mohammed Jaffar AliEmail author
Original Paper

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.

Keyword

Immunomagnetic separation Salmonella PCR Multiplex detection Biofunctionalized nanoparticles 

Notes

Acknowledgment

We acknowledge Central Instrumentation Facility, Pondicherry University for SEM imaging. We thank Dr. Vaidehi, SMF Hospital for providing the clinical isolates. BP thanks Council of Scientific and Industrial Research (CSIR) for the award of Senior Research Fellowship. This work was partially supported by KBC Research Foundation.

Supplementary material

604_2013_1052_MOESM1_ESM.doc (1.4 mb)
ESM 1 (DOC 1414 kb)

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Copyright information

© Springer-Verlag Wien 2013

Authors and Affiliations

  • Padmavathy Bakthavathsalam
    • 1
  • Vinoth Kumar Rajendran
    • 1
  • Uttara Saran
    • 1
  • Suvro Chatterjee
    • 1
  • Baquir Mohammed Jaffar Ali
    • 2
    Email author
  1. 1.AU-KBC Research CentreM.I.T Campus of Anna UniversityChennaiIndia
  2. 2.Centre for Green Energy TechnologyPondicherry UniversityPuducherryIndia

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