3 Biotech

, 9:425 | Cite as

Rapid detection of Salmonella enterica in raw milk samples using Stn gene-based biosensor

  • Kritika Saini
  • Ankur Kaushal
  • Shagun Gupta
  • Dinesh KumarEmail author
Original Article


In this study, a DNA-based nanosensor using specific NH2 labeled single standard probe was developed against stn gene of Salmonella enterica in milk samples. The single-stranded DNA probe was immobilized on carboxylated multiwalled carbon nanotube and gold nanoparticle (c-MWCNT/AuNP) electrode using 1-Ethyl-3-(3-dimethylaminopropyl) carbodiimide (EDC): N-hydroxy succinimide-based cross-linking chemistry. Electrochemical characterization was performed using cyclic voltammetry (CV) and Differential Pulse Voltammetry (DPV) techniques. The electrode surface at each step of fabrication was characterized using scanning electron microscopy. The sensitivity and lower limit of detection were found to be 728.42 (μA/cm2)/ng and 1.8 pg/6 μl (0.3 pg/ml), respectively, with regression coefficient (R2) of 0.843 using DPV. The sensor was further validated using raw and artificial milk samples, and results were compared with conventional methods of detection. The developed sensor was found to be highly sensitive and stable up to 6 months, with only 10% loss of initial peak current in CV analysis on storage at 4 °C.


Salmonella enterica Foodborne illness stn gene Electrochemical biosensor 



The authors are thankful to Shoolini University, Solan, Himachal Pradesh for providing the facility to carry out the present research work. In addition, we would like to acknowledge scientific writing cell for the support provided towards language editing of the manuscript.

Compliances with ethical standards

Conflict of interest

There is no conflict of interest for authorship or related to any other context between authors.

Ethical standards

The authors have complied and worked within standard ethical norms.


  1. Alocilja EC, Zhang D, Shi C (2013) AuNP-DNA biosensor for rapid detection of Salmonella enterica serovar enteritidis. ACS Symp Ser 1143:43–53. CrossRefGoogle Scholar
  2. Das R, Sharma MK, Rao VK (2014) An electrochemical genosensor for Salmonella typhi on gold nanoparticles-mercaptosilane modified screen-printed electrode. J Biotechnol 188:9–16. CrossRefPubMedGoogle Scholar
  3. Dash SK, Sharma M, Khare S, Kumar A (2014) Carbon-mercaptooctadecane/carboxylated multi-walled carbon nanotubes composite based genosensor for detection of bacterial meningitis. Indian J Microbiol 54(2):170–177. CrossRefPubMedGoogle Scholar
  4. Díaz-Serrano M, Rosado A, Del Pilar J (2011) A Polymer-based electrochemical DNA biosensor for Salmonella: preparation, characterization, and calibration. Electroanal 23(8):1830–1841. CrossRefGoogle Scholar
  5. Dong J, Zhao H, Xu M (2013) Label-free electrochemical impedance immunosensor based on AuNPs/PAMAM-MWCNT-Chi nanocomposite modified glassy carbon electrode for detection of Salmonella typhimurium in milk. Food Chem 141(3):1980–1986. CrossRefPubMedGoogle Scholar
  6. Hendriksen RS, Vieira AR, Karlsmose S et al (2011) Global monitoring of Salmonella serovar distribution from the World Health Organization Global Foodborne Infections Network Country Data Bank: results of quality assured laboratories from 2001 to 2007. Foodborne Pathog Dis 8(8):887–900. CrossRefPubMedGoogle Scholar
  7. Jia F, Duan N, Wu S et al (2016) Impedimetric Salmonella aptasensor using a glassy carbon electrode modified with an electrode deposited composite consisting of reduced graphene oxide and carbon nanotubes. Microchim Acta 183(1):337–344. CrossRefGoogle Scholar
  8. Kaushal A, Kumar D, Khare S, Kumar A (2012) speB gene as a specific genetic marker for early detection of rheumatic heart disease in human. Mol Cell Biol 58(1):50–54. CrossRefGoogle Scholar
  9. Li Q, Cheng W, Zhang D et al (2012) Rapid and sensitive strategy for Salmonella detection using an InvA gene-based electrochemical DNA sensor. Int J Electrochem Sci 7(1):844–856Google Scholar
  10. Ma X, Jiang Y, Jia F et al (2014) An aptamer-based electrochemical biosensor for the detection of Salmonella. J Microbiol Methods 98:94–98. CrossRefPubMedGoogle Scholar
  11. Nagraik R, Kaushal A, Gupta S et al (2019) Optimized DNA-based bioassay for Leptospira interrogans detection: a novel platform for leptospirosis diagnosis. 3 Biotech 9:284–291. CrossRefPubMedGoogle Scholar
  12. Ozdemir K, Acar S (2014) Plasmid profile and pulsed-field gel electrophoresis analysis of Salmonella enterica isolates from humans in Turkey. PLoS One 9(5):e95976. CrossRefPubMedPubMedCentralGoogle Scholar
  13. Parvej MS, Nazir KH, Rahman MB (2016) Prevalence and characterization of multi-drug resistant Salmonella enterica serovar Gallinarum biovar Pullorum and Gallinarum from chicken. Vet World 9(1):65–70. CrossRefPubMedPubMedCentralGoogle Scholar
  14. Pashazadeh P, Mokhtarzadeh A, Hasanzadeh M (2016) Recent advances in materials and methods for sensing Salmonella infections. Biosens Bioelectron 87:1050–1064. CrossRefPubMedGoogle Scholar
  15. Savas S, Ersoy A, Gulmez Y et al (2018) Nanoparticle enhanced antibody and DNA biosensors for sensitive detection of Salmonella. Materials 11:1541. CrossRefPubMedCentralGoogle Scholar
  16. Silva Da, Ghica ME, Brett CM (2018) Gold nanoparticle decorated multiwalled carbon nanotube modified electrodes for the electrochemical determination of theophylline. Anal Methods 10(47):5634–5642. CrossRefGoogle Scholar
  17. Singh S, Kaushal A, Khare S et al (2014) Mga genosensor for early detection of human rheumatic heart disease. Appl Biochem Biotechnol 173:228–238. CrossRefPubMedGoogle Scholar
  18. Taha EG, Mohamed A, Srivastava KK et al (2010) Rapid detection of Salmonella in chicken meat using immunomagnetic separation, CHROMagar, ELISA and real-time polymerase chain reaction (RT-PCR). Int J Poult Sci 9:831–835. CrossRefGoogle Scholar
  19. Xu H, Lee HY, Ahn J (2010) Growth and virulence properties of biofilm-forming Salmonella enterica serovar Typhimurium under different acidic conditions. Appl Environ Microbiol 76(24):7910–7917. CrossRefPubMedPubMedCentralGoogle Scholar
  20. Zhu L, Zhao R, Wang K et al (2008) Electrochemical behaviors of methylene blue on DNA modified electrode and its application to the detection of PCR product from NOS sequence. Sens J 8(9):5649–5660. CrossRefGoogle Scholar

Copyright information

© King Abdulaziz City for Science and Technology 2019

Authors and Affiliations

  1. 1.Shoolini University of Biotechnology and Management SciencesDistrict SolanIndia
  2. 2.Amity UniversityGurgaonIndia

Personalised recommendations