Use of duplex polymerase chain reaction (duplex-PCR) technique to identify bovine and water buffalo milk used in making mozzarella cheese

2001 ◽  
Vol 68 (4) ◽  
pp. 689-698 ◽  
Author(s):  
STEFANO REA ◽  
KOICHI CHIKUNI ◽  
RAFFAELLA BRANCIARI ◽  
RAM SUKASI SANGAMAYYA ◽  
DAVID RANUCCI ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Species Identification ◽  
Water Buffalo ◽  
Bovine Milk ◽  
Buffalo Milk ◽  
Animal Origin ◽  
Duplex Pcr ◽  
Mozzarella Cheese ◽  
Chain Reaction ◽  
Polymerase Chain

Molecular biology techniques have been used for species identification in food of animal origin in relatively recent years. A polymerase chain reaction (PCR) based method, the multiplex PCR, was recently applied to species identification in meat and meat products. It allows co-amplification of separate regions of a single gene or specific fragments, each typical of a different animal species in a single PCR reaction, using different pairs of primers in the same reaction mix. In the present paper, the duplex-PCR technique is proposed to identify bovine and water buffalo DNA in a single PCR assay in milk and mozzarella cheese (a typical Italian cheese, originally made from pure water buffalo milk). Because of its lower cost, undeclared bovine milk is added to water buffalo milk for making different kinds of mozzarella cheese. The results of this experiment indicate the applicability of this method, which showed an absolute specificity for the two species and a high sensitivity even down to low DNA concentrations (1 pg). In bovine and water buffalo mixtures of both milk and mozzarella cheese, the minimum concentration tested was 1% of bovine in water buffalo milk and water buffalo in bovine milk. The importance of the somatic cell content in raw milk is also discussed with special reference to the evaluation of mixtures (milk or cheese) of the two species.

scholarly journals Droplet Digital PCR (ddPCR) Analysis for the Detection and Quantification of Cow DNA in Buffalo Mozzarella Cheese

Animals ◽  
2021 ◽  
Vol 11 (5) ◽  
pp. 1270
Author(s):  
Anna Cutarelli ◽  
Andrea Fulgione ◽  
Pasquale Fraulo ◽  
Francesco Paolo Serpe ◽  
Pasquale Gallo ◽  
...  
Keyword(s):  
Digital Pcr ◽  
Buffalo Milk ◽  
Cow Milk ◽  
Mozzarella Cheese ◽  
Chain Reaction ◽  
New Methods ◽  
Protected Designation Of Origin ◽  
Commission Regulation ◽  
Polymerase Chain ◽  
Digital Polymerase Chain Reaction

Buffalo mozzarella cheese is one of the most appreciated traditional Italian products and it is certified as a Protected Designation of Origin (PDO) product under the European Commission Regulation No. 1151/2012. It is obtained exclusively from buffalo milk. If made from cow milk, or a mixture of buffalo and cow milk, buffalo mozzarella cheese does not qualify as a PDO product. In order to maximize their profits, some producers market buffalo mozzarella that also contains cow milk as a PDO product, thus defrauding consumers. New methods for revealing this fraud are therefore needed. One such method is the droplet digital Polymerase Chain Reaction (ddPCR). Thanks to its high precision and sensitivity, the ddPCR could prove an efficacious means for detecting the presence of cow milk in buffalo mozzarella cheese that is marketed as a PDO product. ddPCR has proved able to detect the DNA of cow and/or buffalo milk in 33 buffalo mozzarella cheeses labelled as PDO products, and experimental evidence could support its application in routine analyses.

scholarly journals Application of a multiplex polymerase chain reaction (mPCR) assay to detect fraud by substitution of bovine meat cuts with water buffalo meat in Northern Brazil

2019 ◽  
Vol 17 (1) ◽  
pp. 790-795 ◽  
Author(s):  
Vanderson Vasconcelos Dantas ◽  
Gabrielle Virginia Ferreira Cardoso ◽  
Wanessa Shuelen Costa Araújo ◽  
Andrey Carlos do Sacramento de Oliveira ◽  
Andreia Silva da Silva ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Multiplex Polymerase Chain Reaction ◽  
Water Buffalo ◽  
Chain Reaction ◽  
Buffalo Meat ◽  
Northern Brazil ◽  
Polymerase Chain ◽  
Mpcr Assay

Extraction of microsporidial DNA from modified trichrome-stained clinical slides and subsequent species identification using PCR sequencing

Parasitology ◽  
2008 ◽  
Vol 135 (6) ◽  
pp. 701-703 ◽  
Author(s):  
K. S. CHAN ◽  
T. H. KOH
Keyword(s):  
Polymerase Chain Reaction ◽  
Species Identification ◽  
Environmental Samples ◽  
Species Level ◽  
Molecular Techniques ◽  
Chain Reaction ◽  
Clinical Specimens ◽  
Polymerase Chain ◽  
Pcr Conditions ◽  
Hiv Negative

SUMMARYMolecular techniques involving polymerase chain reaction (PCR) and sequencing provide a relatively simple and objective means of identifying microsporidia to species level. We modified previously described methods of DNA extraction and PCR conditions for identification of microsporidia from museum slides, clinical specimens and environmental samples and successfully identifiedVittaforma corneaein 11 out of 13 cases of microsporidial infection from used trichrome-stained slides of corneal scrapings from HIV-negative patients with keratoconjunctivitis.

scholarly journals Detection and enumeration of Staphylococcus aureus from bovine milk samples by real-time polymerase chain reaction

2013 ◽  
Vol 96 (11) ◽  
pp. 6955-6964 ◽  
Author(s):  
B.G. Botaro ◽  
C.S. Cortinhas ◽  
L.V. Março ◽  
J.F.G. Moreno ◽  
L.F.P. Silva ◽  
...  
Keyword(s):  
Staphylococcus Aureus ◽  
Polymerase Chain Reaction ◽  
Real Time ◽  
Bovine Milk ◽  
Chain Reaction ◽  
Milk Samples ◽  
Polymerase Chain

Survival of Salmonellae in Pasteurized, Refrigerated Calcium-Fortified Orange Juice

2001 ◽  
Vol 64 (9) ◽  
pp. 1299-1304 ◽  
Author(s):  
MANAN SHARMA ◽  
LARRY R. BEUCHAT ◽  
MICHAEL P. DOYLE ◽  
JINRU CHEN
Keyword(s):  
Polymerase Chain Reaction ◽  
Orange Juice ◽  
Storage Period ◽  
Animal Origin ◽  
Calcium Citrate ◽  
Chain Reaction ◽  
Polymerase Chain ◽  
Acidic Environments ◽  
Number Of Cells ◽  
Salmonella Heidelberg

Studies were done to determine the survival of salmonellae in orange juice as affected by fortification with calcium. Four brands of commercially pasteurized orange juice fortified with calcium (350 mg/240-ml serving) and nonfortified juice were inoculated separately with three types of inocula: strains of Salmonella Muenchen (inoculum 1), serotypes of human and animal origin (inoculum 2), and isolates from raw produce- and juice-associated outbreaks (inoculum 3). Juice inoculated with populations of 6.6 to 7.0 log10 CFU of Salmonella per ml was held at 4°C for up to 32 days. The number of cells of inoculum 1 that survived in juice fortified with calcium lactate/tricalcium phosphate (CaL/TCP) was significantly lower (P ≤ 0.05) (2.80 log10 CFU/ml) than in nonfortified juice (3.50 log10 CFU/ml) after 32 days' storage. Death of salmonellae in inocula 1 and 2 was less in juice fortified with TCP (3.21 and 3.33 log10 CFU/ml, respectively) than in the nonfortified juice (3.75 and 4.15 log10 CFU/ml, respectively). During the 32-day storage period, populations in inocula 1 and 3 showed significantly less inactivation (2.62 and 3.12 log10 CFU/ml, respectively) in juice fortified with calcium citrate (CC) than in nonfortified juice (3.14 and 3.60 log10 CFU/ml, respectively).There were no significant differences in the survival of Salmonella in juice fortified with calcium citrate malate (CCM) and nonfortified juice. Polymerase chain reaction (PCR) typing of randomly selected Salmonella colonies revealed that Salmonella Heidelberg in inoculum 2 and Salmonella Baildon and Salmonella Poona in inoculum 3 were the most prevalent at the end of the 32-day storage period at 4°C, suggesting that serotypes selected for use in inocula differed in tolerance to acidic environments. This study reveals that the form of calcium used to fortify orange juice may affect the survival of Salmonella.

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