Selective identification of specific aptamers for the detection of non-typhoidal salmonellosis in an apta-impedimetric sensing format

Abstract

The authors report on an aptamer-based method for the detection of S. Typhimurium. The aptamers were identified by using a modified cell-based SELEX method (cell-SELEX) by adopting an alternating negative-positive selection strategy. Each cross-reactive population was used separately, with a focus on generating strain-specific aptamers in order to enhance selectivity. The aptamers were characterized and chemically modified with thiol groups at the 5′ end which enables covalent binding to gold nanoparticles on a screen printed carbon electrode (GNP-SPCE). The aptamer-modified GNP-SPCE was subsequently applied to impedimetric sensing of S. Typhimurium in water and food samples. Response is linear in the 10 to 105 CFU⋅mL−1 concentration range, and the limit of detection is ~10 CFU⋅mL−1. The assay can distinguish S. Typhimurium from S. Typhi and has been validated in water and spiked egg samples.

A modified Cell-SELEX approach for screening species specific aptamer for the impedimetric detection of non-typhoidal Salmonellosis.

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Acknowledgements

The authors thank DBT, India (Grant No. BT/IN/FINNISH/09/CRS/2013) for providing the financial support for this project. Authors also thank Dr. Priyanka Sabherwal and Munish Shorie for the technical support, and Mr. Deepak Bhatt for confocal studies.

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Correspondence to Praveen Rishi or C. Raman Suri.

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Pathania, P., Sharma, A., Kumar, B. et al. Selective identification of specific aptamers for the detection of non-typhoidal salmonellosis in an apta-impedimetric sensing format. Microchim Acta 184, 1499–1508 (2017). https://doi.org/10.1007/s00604-017-2098-2

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Keywords

  • S. Typhimurium
  • Modified cell-SELEX
  • Gold nanoparticles
  • Impedimetry
  • Aptasensor
  • Microbial pathogen