A sensitive DNA-based fluorometric method for milk authenticity of dairy products based on spectrally distinct microspheres

Kounelli, Maria L.; Kalogianni, Despina P.

doi:10.1007/s00217-017-2882-6

Cited by 16 publications

(9 citation statements)

References 45 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Kounelli and Kalogianni [143] developed a DNA-based method that relied on hybridization of species-specific oligonucleotide on the surface of fluorescent microspheres, followed by flow cytometry analysis. The method consisted of DNA amplification with species-specific labeled primers targeting the cytb gene of each species (cow, sheep and goat), followed by hybridization of the single-strand biotinylated PCR products with species-specific oligonucleotide probes, carrying a NH 2 group at the 5 -end, which were attached to the surface of three different sets of carboxylated microspheres.…”

Section: (Bio)sensorsmentioning

confidence: 99%

Animal Species Authentication in Dairy Products

Mafra

Honrado

Amaral

2022

Foods

View full text Add to dashboard Cite

Milk is one of the most important nutritious foods, widely consumed worldwide, either in its natural form or via dairy products. Currently, several economic, health and ethical issues emphasize the need for a more frequent and rigorous quality control of dairy products and the importance of detecting adulterations in these products. For this reason, several conventional and advanced techniques have been proposed, aiming at detecting and quantifying eventual adulterations, preferentially in a rapid, cost-effective, easy to implement, sensitive and specific way. They have relied mostly on electrophoretic, chromatographic and immunoenzymatic techniques. More recently, mass spectrometry, spectroscopic methods (near infrared (NIR), mid infrared (MIR), nuclear magnetic resonance (NMR) and front face fluorescence coupled to chemometrics), DNA analysis (real-time PCR, high-resolution melting analysis, next generation sequencing and droplet digital PCR) and biosensors have been advanced as innovative tools for dairy product authentication. Milk substitution from high-valued species with lower-cost bovine milk is one of the most frequent adulteration practices. Therefore, this review intends to describe the most relevant developments regarding the current and advanced analytical methodologies applied to species authentication of milk and dairy products.

show abstract

Section: (Bio)sensorsmentioning

confidence: 99%

Animal Species Authentication in Dairy Products

Mafra

Honrado

Amaral

2022

Foods

View full text Add to dashboard Cite

show abstract

“…There are many methods have been used for identification of species origin of raw milk as chemical, immunological, and molecular techniques. There are several molecular techniques namely, PCR-RFLP [ 18 ], multiplex PCR [ 14 ] and Real-time PCR [ 21 ], and DNA based fluorometric method [ 22 ]. These PCR techniques can be used to differentiate between the closed related species.…”

Section: Discussionmentioning

confidence: 99%

Species adulteration in raw milk samples using polymerase chain reaction-restriction fragment length polymorphism

Ewida¹,

El-Magiud²

2018

Vet World

View full text Add to dashboard Cite

Background and Aim:Milk adulteration is pivotal because it leads to worse effects in public health as human adverse reactions with clinical signs ranged from gastrointestinal signs to anaphylactic shock. This study was carried out to estimate the prevalence of adulteration in buffalo’s milk sold in Assiut City, Egypt.Materials and Methods:A total of 50 raw buffalo’s milk samples were collected and examined for adulteration by addition of cow’s milk. The examination carried out by applying polymerase chain reaction-restriction fragment length polymorphism technique using cytochrome b (cyt b) gene primers and Hinf I enzymes. The size of target gene was 360 bp in both animal species and amplicon can be digested using Hinf I enzyme, this restriction enzyme divided the essential band to clear three bands at 360, 210, and 150 bp in cows’ milk, while, the enzyme could not be cleaved the amplicon in buffalo’s samples.Results:The obtained results cleared that the incidence of adulteration of buffalo’s milk very high percentage reaches 90%.Conclusion:It could be concluded that the raw buffalo’s milk sold in Assiut City subject to fraudulent practice and thus can lead to public health hazards.

show abstract

“…In 2017, Kounelli et al developed a sensitive method based on a fluorometric DNA hybridization assay on polystyrene microspheres that contain carboxyl groups on their surface, coupled to specific probes for bovine, sheep and goat species. The method was centred in the amplification of mitochondrial cytochrome b gene from those species, allowing detection limits as lower as 0.01% of bovine milk in binary mixtures of bovine/goat yogurts and 0.05% proportion in bovine/sheep yogurt mixtures (Kounelli & Kalogianni, 2017). Beltramo et al applied a biochip (LCD array) for cattle, goat, and buffalo species identification in milk admixtures, using a method based on the hybridization of biotinylated amplicons specific to cytochrome b oxidase gene of all three species that allowed simultaneous detection of several targets and revealed a sensitivity of 0.1% (Beltramo et al, 2017).…”

Section: Species-differentiation Techniquesmentioning

confidence: 99%

DNA-based approaches for dairy products authentication: A review and perspectives

Baptista

Cunha

Domingues

2021

Trends in Food Science & Technology

View full text Add to dashboard Cite

Background: Food fraud has become an increasingly worldwide problem mainly driven by rapid innovation in the food sector and constantly changing consumer's choices. This led to an increased necessity to improve/establish reliable authentication processes, resulting in the replacement of protein-based techniques for dairy products authentication by higher sensitive and reproducible DNA-based methods. Most used molecular methods for dairy products authentication include PCR, Real-time PCR, multiplex PCR, and PCR-RFLP. Despite the several molecular methods available for species/breeds differentiation in dairy products, there is a need for the development of more efficient and reliable molecular tools. Scope and approach: In this review, traditional and more recent DNA-based methods for dairy products authentication are discussed and analysed. Moreover, the increasingly important role of bioinformatic tools for analysis of large amount of data and for the development of DNA markers is also discussed. Key findings and conclusions: DNA quality is one of the major factors affecting molecular-based dairy products authentication, which can be influenced by the manufacturing process, extraction method employed, chemical composition of the food matrix, among others. PCR-based methods continue to be the most important and successfully used for dairy products authentication. The DNA markers chosen for species/breed detection are an important factor for PCR success. Although there are several molecular methods for the detection of adulterant species, there is still an unmet demand for methods to detect adulterant breeds. Available public databases and bioinformatics revolutionized the analysis of large amount of data and will be pivotal for the development of effective DNA markers.

show abstract

A sensitive DNA-based fluorometric method for milk authenticity of dairy products based on spectrally distinct microspheres

Cited by 16 publications

References 45 publications

Animal Species Authentication in Dairy Products

Animal Species Authentication in Dairy Products

Species adulteration in raw milk samples using polymerase chain reaction-restriction fragment length polymorphism

DNA-based approaches for dairy products authentication: A review and perspectives

Contact Info

Product

Resources

About