title: Simultaneous detection of cow and buffalo species in milk from China, India, and Pakistan using multiplex real-time PCR.
abstract: Asian countries are major producers of cow and buffalo milk. For quality and authenticity purposes, a multiplex real-time PCR assay was developed to specifically and simultaneously detect DNA from these 2 bovine species. Targeting the cytochrome b gene of mitochondrial DNA, common PCR primers amplified a 105-bp fragment, and 2 fluorescent probes specific to either cow or buffalo were designed for their identification. Specificity was successfully tested on 6 other species, including sheep and goat, and sensitivity reached 1% of cow DNA in buffalo DNA and vice versa. As an evaluation, the method was tested using 119 freeze-dried Asian milk samples from regional industrial milk facilities. Although these samples did not cover the entire Asian zone, the multiplex assay indicated that approximately 20% of the samples (mainly from India) showed high levels of cross-contamination of cow milk by buffalo milk, and vice versa. Fast, sensitive, and straightforward, this method is fit-for-purpose for the authenticity control of Asian milk.
authors.affiliation: Quality and Safety Department, Nestlé Research Center, PO Box 44, 1000 Lausanne 26, Switzerland. geoffrey.cottenet@rdls.nestle.com
authors.forename: G
authors.initials: G
authors.lastname: Cottenet
authors.forename: C
authors.initials: C
authors.lastname: Blancpain
authors.forename: P-A
authors.initials: PA
authors.lastname: Golay
mesh_terms: D000818:Animals; D002020:Buffaloes / Q000235:genetics; D002417:Cattle / Q000235:genetics; D002681:China; D004247:DNA / Q000235:genetics; D007194:India; D008892:Milk / Q000737:chemistry* / Q000592:standards; D010154:Pakistan; D060888:Real-Time Polymerase Chain Reaction / Q000662:veterinary*; D012680:Sensitivity and Specificity
chemical_list: D004247:DNA