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

Abstract

The adulteration of meat products can affect the confidence of consumers and the market, leading to negative impacts on the economy. Accordingly, product authenticity has become an important issue in modern society. Therefore, our study aimed to optimize the extraction of DNA from meat and use multiplex polymerase chain reaction (mPCR) to determine the incidence of fraud by substitution of bovine meat cuts with water buffalo meat in the states of Pará and Amapá, Northern Brazil. The mPCR protocol used primers that amplify sequences of 346 base pairs of bovine DNA and 220 base pairs of buffalo DNA. To assess the sensitivity of the technique, a standardized PCR assay was performed using the template DNA extracted and diluted from 10−0 to 10−10 in PCR-grade water. Next, 161 samples of meat cuts marketed as bovine (rump) origin were collected in the states of Pará and Amapá, Northern Brazil. The mPCR assays demonstrated good specificity of the primers used. The sensitivity test amplified bovine and buffalo DNA fragments down to the 10−2 dilution. The results demonstrated fraud by substitution of beef by water buffalo meat in 21.7% of samples, demonstrating that this act does occur intentionally for economic gains.

Keywords

• dna extraction
• multiplex pcr
• species identification
• food fraud