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Rapid Detection Device for Salmonella typhi in Milk, Juice, Water and Calf Serum

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Abstract

A limit of detection of 200 CFU/mL of Salmonella typhi spiked in various sample matrices were achieved in 30 min. The sample matrices were raw/unprocessed milk, commercially available milk, juice from packed bottles, fresh juice from carts, potable water, turbid water and calf serum. The complete protocol comprised of three steps: (a) cell lysis (b) nucleic acid amplification and (c) an in situ optical detection. The cell lysis was carried out using a simple heating based protocol, while the loop-mediated isothermal amplification of DNA was carried out by an in-house designed and fabricated system. The developed system consists of an aluminum block fitted with two cartridge heaters along with a thermocouple. The system was coupled to a light source and spectrometer for a simultaneous in situ detection. Primers specific for STY2879 gene were used to amplify the nucleic acid sequence, isolated from S. typhi cells. The protocol involves 15 min of cell lysis and DNA isolation followed by 15 min for isothermal amplification and simultaneous detection. No cross-reactivity of the primers were observed at 106 CFU/mL of Escherichia coli, Vibrio cholerae, Salmonella typhimurium, Salmonella paratyphi A, Pseudomonas aeruginosa, Bacillus cereus, Lysteria monocytogenes, Clostridium botulinum, Staphylococcus aureus and Salmonella havana. In addition, the system was able to detect S. typhi of 200 CFU/mL in a concoction of 106 CFU/mL of E. coli, 106 CFU/mL of V. cholerae, and 106 CFU/mL of hepatocyte-derived cellular carcinoma HUH7 cells. The proposed rapid diagnostic system shows a promising future in the field of food and medical diagnostics.

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Acknowledgements

The authors would also like to acknowledge Professor Arti Kapil of All India Institute of Medical Sciences for valuable inputs.

Funding

The authors would like to acknowledge the funding agencies for their continued support: Department of Science and Technology (YSS/2014/000880, and IDP/MED/05/2014), Indo-German Science and Technology Centre (IGSTC/Call 2014/Sound4All/24/2015-16), Naval research board (NRB/4003/PG/359), BIRAC, Department of Biotechnology (BIRAC/BT/AIR0275/PACE-12/17).

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Authors and Affiliations

  1. Device Testing Laboratory, Centre for Biomedical Engineering, Indian Institute of Technology Delhi, New Delhi, 110016, India

    Avinash Kaur, Ritu Das, Mayank Rai Nigam & Dinesh Kalyanasundaram

  2. Department of Biochemical and Biotechnology, Indian Institute of Technology Delhi, New Delhi, 110016, India

    Ravikrishnan Elangovan

  3. AmpliGene India Biotech Private Limited, Ahmedabad, India

    Deepal Pandya

  4. Lab-on-a-chip and Biosensors Laboratory, Centre for Biomedical Engineering, Indian Institute of Technology Delhi, New Delhi, 110016, India

    Sandeep Jha

  5. Department of Biomedical Engineering, All India Institute of Medical Sciences, New Delhi, 110029, India

    Sandeep Jha & Dinesh Kalyanasundaram

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  1. Avinash Kaur
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  2. Ritu Das
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  3. Mayank Rai Nigam
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  4. Ravikrishnan Elangovan
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  7. Dinesh Kalyanasundaram
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Correspondence to Sandeep Jha or Dinesh Kalyanasundaram.

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Conflicts of interest

AK declares that she has no conflict of interest. RD declares that she has no conflict of interest. MRN declares that he has no conflict of interest. RE declares that he has no conflict of interest. DP declares that she has no conflict of interest. SJ declares that he has no conflict of interest. DK declares that he has no conflict of interest.

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The study does not involve any human samples and there is no ethical clearance required for this work.

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Kaur, A., Das, R., Nigam, M.R. et al. Rapid Detection Device for Salmonella typhi in Milk, Juice, Water and Calf Serum. Indian J Microbiol 58, 381–392 (2018). https://doi.org/10.1007/s12088-018-0730-4

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  • DOI: https://doi.org/10.1007/s12088-018-0730-4

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