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High resolution melting is a useful tool to detect animal species sources of various milk types

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Abstract

Accurate identification of animal species sources in milk have become quite important due to adulteration of high-priced milk types in the dairy industry. To date, milk identification methods have mainly depended on biochemical properties or physical properties detected by spectroscopic methods. The current study aimed to develop an easy to use and sensitive DNA-based High resolution melting (HRM) assay to identify animal species and detect cross-adulteration of water buffalo, bovine, goat, sheep, camel and donkey milks. HRM compatible designed primer set, targeted mitochondrial region, successfully amplified the specific targeted region for six animal species DNA and showed a high degree of specificity based on nucleotide variations. Capillary electrophoresis analysis validated the specific amplicons and determined the amplicon lengths as 114 bp for bovine, goat, sheep, and camel, 115 bp for water buffalo, and 121 bp for donkey. HRM analysis showed a clear discrimination for water buffalo-bovine, camel-bovine and donkey-bovine adulteration down to 0.5%, and goat-sheep adulteration down to 1% in the milk admixtures. The efficacy of the method was also confirmed by its standard curve with a very high correlation coefficient In conclusion, the designed HRM assay allows for the rapid, sensitive and cost-effective authentication of milk and dairy products.

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Data availability

All data generated or analysed during this study are included in this published article (and its supplementary information files). The nucleotide alignment data was given as supplementary file.

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Not applicable.

Abbreviations

HRM:

High resolution melting

PCR:

Polymerase chain reaction

NIR:

Near infrared

dsDNA:

Double-stranded DNA

SFS:

Synchronous fluorescent spectroscopy

W:

Water buffalo

B:

Bovine

G:

Goat

S:

Sheep

C:

Camel

D:

Donkey

WB:

Water buffalo–Bovine

GS:

Goat–Sheep

CB:

Camel–Bovine

DB:

Donkey–Bovine

CE:

Capillary electrophoresis

GCPs:

Genotyping confidence percentages

Cq :

Quantification cycle

Ct :

Melting point

SD:

Standard deviation

DNA:

Deoxyribonucleic acid

bp:

Base pair

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Acknowledgements

We thank anonymous reviewers for their improvements on the manuscript.

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There is no funding available for this publication.

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Authors

Contributions

KH: Investigation, Formal Analysis, writing original draft; KH&MB: Conceptualization, methodology, validation, writing—review & edit.

Corresponding author

Correspondence to Kaan Hürkan.

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Hürkan, K., Bulut, M. High resolution melting is a useful tool to detect animal species sources of various milk types. J Food Sci Technol 60, 1612–1620 (2023). https://doi.org/10.1007/s13197-023-05705-3

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  • DOI: https://doi.org/10.1007/s13197-023-05705-3

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