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Development of an on-site lateral flow immune assay based on mango leaf derived colloidal silver nanoparticles for rapid detection of Staphylococcus aureus in milk

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Abstract

In order to ensure food safety, screening food samples for the presence of pathogens has been categorised as a legal testing item throughout the globe. One of the most prevalent zoonotic bacteria transmitted through dairy milk is Staphylococcus aureus. Given the limitations of the conventional detection methods, in the current study we desigined a competitive lateral flow immune assay (LFIA) using colloidal silver nanoparticles derived from mango leaves for the detection of Staphylococcus aureus in cow milk. SpA, a recombinant protein of Staphylococcus aureus, was used to raised hyperimmune sera used for developing the assay followed by conjugation with the synthesized nanoparticles. To increase the specificity of the assay, the milk samples were prenriched with selective agar exclusively require for Staphyloccocus aureus. The assay was found to be completed within 7–8 h by observing test and control lines in LFIA strips. The developed assay was found to specifically detect the bacteria as low as 1000 cfu/ml of milk samples. With a total 230 number of raw and clinical mastitis milk samples, the assay was validated and achieved relative accuracy, specificity, and sensitivity values of 97.39, 98.03, and 96.1%, respectively. The developed LFIA, which uses economically feasible and stable silver nanoparticles derived from mango leaves, has the potential for routine screening of milk samples for the presence of Staphylococcus aureus, especially in low-resource settings, allowing for early diagnosis, which facilitates effective treatment for the dairy animals and prevents the transmission of the disease in consumers.

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Data availability

Data sharing not applicable to this article as no datasets were generated or analysed during the current study.

Code availability

Not applicable.

Abbreviations

AgNPs:

Silver nano particles

LFIA:

Lateral flow immune assay

SpA:

Staphylococcus aureus Recombinant protein A

ELISA:

Enzyme linked immune sorbent assay

FTIR:

Fourier transform infrared

XRD:

X-Ray diffraction

CFU:

Colony forming unit

LOD:

Limit of detection

PPV:

Positive predictive value

NPV:

Negative predictive value

AC:

Relative accuracy

SP:

Specificity

SE:

Sensitivity

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Acknowledgements

Authors acknowledge the financial support received from Indian Council of Agricultural Research, New Delhi, India as ICAR-Post Doctoral Fellowship to RD during this study period. Authors are also thankful to Science and Engineering Research Board, Department of Science and Technology, Government of India for the financial assistant to SD for the ongoing project code EMR/2017/004602. Authors are thankful to Director, ICAR-National Dairy Research Institute, Karnal, Haryana, India for providing necessary facilities to conduct the experiment.

Funding

Indian Council of Agricultural Research, New Delhi, India as ICAR-Post Doctoral Fellowship and Science and Engineering Research Board, Department of Science and Technology, Government of India.

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Authors and Affiliations

  1. Animal Genomics Laboratory, Animal Biotechnology Center, ICAR-National Dairy Research Institute, Karnal-1320021, Haryana, India

    Rajib Deb, Parul Chaudhary, Prassana Pal, Rahul Singh Tomar, Mayank Roshan,  Parmanand, Ashutosh Ludri & Sachinandan De

  2. ICAR-National Research Center On Pig, Guwahati, Assam, India

    Rajib Deb & Vivek Kumar Gupta

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  1. Rajib Deb
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  9. Sachinandan De
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Contributions

RD: conceptualized the idea, performing laboratory bench works, writing manuscript. PC, PP: antibody raising. RST: nanoparticle synthesis. MR, Parmanand, AL: bacterial cultures. VKG: Manuscript revision. SD: Monitoring the research work and revision of the final manuscript.

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Correspondence to Rajib Deb.

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The funding agency has been acknowledged, and all authors involved in the research work have been listed in the authors section. We declare that we have no competing interests.

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Deb, R., Chaudhary, P., Pal, P. et al. Development of an on-site lateral flow immune assay based on mango leaf derived colloidal silver nanoparticles for rapid detection of Staphylococcus aureus in milk. J Food Sci Technol 60, 132–146 (2023). https://doi.org/10.1007/s13197-022-05598-8

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  • DOI: https://doi.org/10.1007/s13197-022-05598-8

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