Simultaneous detection of cow and buffalo species in milk from China, India, and Pakistan using multiplex real-time PCR |
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Authors: | Cottenet G Blancpain C Golay P-A |
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Affiliation: | Quality and Safety Department, Nestlé Research Center, PO Box 44, 1000 Lausanne 26, Switzerland |
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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. |
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Keywords: | cow milk buffalo milk multiplex real-time polymerase chain reaction |
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