Simplex and duplex PCR assays for species specific identification of cattle and buffalo milk and cheese

De, Sachinandan; Brahma, Biswajit; Polley, Shamik; Mukherjee, Ayan; Banerjee, D. P.; Gohaina, Moloya; Singh, Karan; Singh, Rameswar; Datta, Tirtha Kumar; Goswami, S. L.

doi:10.1016/j.foodcont.2010.09.026

Cited by 48 publications

(35 citation statements)

References 31 publications

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“…1). The results showed the applicability of the primers proposed by De et al (2011) for the identification of cattle and buffalo species and by Dalmasso et al (2004) for the identification of ruminants (in the present study, cattle species).…”

Section: Discussionsupporting

confidence: 62%

“…Nuclear DNA is larger molecule arranged into chromosomes and contains a greater variation in its type of sequences compared to mtDNA. The mtDNA is commonly used for species identification in food analysis (Meyer et al 1994;Matsunaga et al 1999;Girish et al 2004;Che Man et al 2007;Zhang et al 2007;Sahilah et al 2011;De et al 2011) especially cytochrome b (cyt b) gene, 12S rRNA and D loop gene. High copy number of mtDNA is found in the cells and, it remains intact during food processing thereby minimizing DNA degradation and does not contain any introns (Unseld et al 1995).…”

Section: Introductionmentioning

confidence: 99%

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Species identification of cattle and buffalo fat through PCR assay

Vaithiyanathan

Kulkarni

2016

J Food Sci Technol

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A method was standardized to isolate quality DNA from cattle and buffalo fat for species identification using QIAamp DNA stool mini kit. The quality of the DNA was sufficient enough to amplify universal primers viz., mt 12S rRNA and mt 16S rRNA, and species specific D loop primers for cattle and buffalo. The sensitivity of the PCR assay in the species specific D loop primer amplification was with a detection level of 0. 47 ng cattle DNA and 0.23 ng buffalo DNA in simplex and, 0. 47 ng cattle DNA and 0.12 ng buffalo DNA in duplex PCR. It is a potentially reliable method for DNA detection to authenticate animal fat.

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Section: Discussionsupporting

confidence: 62%

Section: Introductionmentioning

confidence: 99%

Species identification of cattle and buffalo fat through PCR assay

Vaithiyanathan

Kulkarni

2016

J Food Sci Technol

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“…quality of DNA, specificity of primers, etc. [12] The main problem for developing the PCR-based approach to ascertain the purity of ghee are highly degraded DNA and recovery of DNA from fat matrices, as fat can potentially inhibit PCR amplification. [22] The main challenge was to isolate amplifiable DNA for PCR reaction from fat matrices.…”

Section: Standardisation Of Pcr Conditions To Increase the Specificitmentioning

confidence: 99%

“…[11] Therefore, DNA-based methodologies are gaining popularity for identifying the origin of food from animal sources (fish, milk, and meat). The DNA-based methodology for identification of species in milk and milk products, [12] meat, [13][14][15] and fish [16] has received important attention in the food industry due to high sensitivity and specificity. However, the DNA-based protocol to detect the addition of body fat and to ascertain the quality of clarified milk (ghee) fat has not been exploited till date.…”

Section: Introductionmentioning

confidence: 99%

A species specific simplex polymerase chain reaction-based approach for detection of goat tallow in heat clarified milk fat (ghee)

Hazra

Sharma

et al. 2017

International Journal of Food Properties

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The detection of milk fat adulteration with foreign fats/oils continues to be a challenge for the dairy industry as well as food testing laboratories. Recently, various chromatographic methods have been developed to detect a low level of vegetable oil in ghee but detection of animal body fat in ghee is still a challenge. In India, goat body fat (goat tallow) adulteration in ghee is a very serious problem. In the present investigation, a rapid species specific polymerase chain reaction (PCR)-based methodology has been standardised to detect goat tallow in heat clarified milk fat (ghee). This standardised PCR-based protocol did not show any false positive results in the genuine ghee (heat clarified butter fat) samples. Adulteration of ghee (both cow and buffalo) with goat tallow at the 10% level could be detected using the standardised PCR-based protocol. The protocol standardised is rapid and convenient to use. ARTICLE HISTORY

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“…A number of DNA-based methods outstand among these procedures, because these techniques have several advantages in contrast to fatty acid analysis and protein-based methods, such as high sensitivity, reproducibility, applicability, and these are cost-effective techniques in species identifi cation practice (DE et al, 2011). The simplest form of PCR-based methods is single-PCR, using one primer pair to detect the presence of specifi ed species (SANTOS et al, 2012).…”

Section: Pcr-single Strand Conformation Polymorphismmentioning

confidence: 99%

Applicability and sensitivity of PCR-SSCP method for milk species identification in cheese

Csikós¹,

Hodžić

Pasic-Juhas

et al. 2016

Acta Alimentaria

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Species identifi cation in food has become a prominent issue in recent years as the importance of consumer protection has increased. DNA-based species identifi cation methods were developed by researchers in the last two decades, as these are reliable, accurate, and low-cost techniques for species identifi cation in raw and processed food products as well. In our study, universal primers were designed to conserved regions of mitochondrial 12S rRNA. Amplicons were heat-denatured and a PCR single strand conformation polymorphism (SSCP) method was developed to identify cattle, buffalo, sheep, and goat DNA. Sensitivity of this technique was tested on DNA mixtures of cattle-sheep, cattle-goat, and cattle-buffalo and the threshold limit of cattle DNA was 5%, 5%, and 3%, respectively. One hundred and fi ve cheeses were purchased and collected from Bosnian and Hungarian farmers, retails, and supermarkets to reveal fraud, 32 percent of them (34 cheeses) were found to be mislabelled by species.

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Simplex and duplex PCR assays for species specific identification of cattle and buffalo milk and cheese

Cited by 48 publications

References 31 publications

Species identification of cattle and buffalo fat through PCR assay

Species identification of cattle and buffalo fat through PCR assay

A species specific simplex polymerase chain reaction-based approach for detection of goat tallow in heat clarified milk fat (ghee)

Applicability and sensitivity of PCR-SSCP method for milk species identification in cheese

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