Simultaneous detection of ovine and caprine DNA in meat and dairy products using triplex TaqMan real‐time PCR

Abstract

Meat and dairy products provide essential nutrients that support growth, functions, immunity, and general human well-being. Xilingol grassland (203,000 km 2) in China is a natural grazing region, known for the free raising of animals for human consumption and its low pollution levels, resulting in the production of high-quality meat and dairy products from sheep and goat, among others. The average annual output of sheep and goat in Xilingol grassland is more than ten million of animals. Nevertheless, some producers and traders deliberately adulterate meat and dairy products with less expensive substitutes, which unbalances market competition and disregards consumer's interests. Further, product authentication is required to manage health risks and to comply with religious norms. Therefore, the development of efficient approaches for product authentication is essential to ensure the purity of food products. Here, we developed a specific and highly sensitive method to evaluate adulteration of meat and milk from sheep and goat based on DNA detection using real-time PCR with TaqMan technology. Distinct analytical PCR-based methods have been developed for DNA detection in animal-derived products such as meat and dairy

Animal Species Authentication in Dairy Products

“…Both systems were applied in the estimation of cow's milk of fresh and ripened model cheeses, with the nuclear systems revealing the highest specificity and quantitative performance. Later on, the same group of researchers developed three triplex real-time PCR methods with TaqMan probes targeting the 12S rRNA gene to simultaneously detect an endogenous control sequence and two species, namely cow and mare [132], cow and goat [133], sheep and goat [134] and camel and cow [135]. The approaches were successfully applied to processed dairy products, achieving high sensitivities down to few pictograms of DNA (Table 3).…”

Rapid detection of Pseudomonas aeruginosa using a DNAzyme‐based sensor

“…Traditional culture methods (Rashno Taee et al., 2014) are highly accurate but are time‐consuming and require specific media and differentiation tests to identify them. Immunological assays (Huang et al., 2020; Lin, Zhou, & Tang, 2017) and polymerase chain reaction (PCR) (Guo et al., 2020) technology based on nucleic acid amplification greatly shortens the duration of identification. However, immunological assays (Krithiga et al., 2016; Shahdordizadeh et al., 2017) suffer from being expensive and complicated sample preparation steps.…”

“…Immunological assays (Huang et al, 2020; and polymerase chain reaction (PCR) (Guo et al, 2020) technology based on nucleic acid amplification greatly shortens the duration of identification. However, immunological assays (Krithiga et al, 2016;Shahdordizadeh et al, 2017) suffer from being expensive and complicated sample preparation steps.…”

Improved triplex real‐time PCR with endogenous control for synchronous identification of DNA from chicken, duck, and goose meat

“…Conventional PCR (Amaral et al., 2015; Martin et al., 2007; Yao et al., 2020), multiplex PCR (Hou et al., 2015; Thanakiatkrai et al., 2019), and real‐time PCR (Furutani et al., 2017; Kesmen et al., 2012; Pegels et al., 2012) are highly specific and efficient methods which have been widely adopted in the detection of meat adulteration. Taq Man real‐time PCR, based on probes labeled by different fluorescent reporters, combines the advantages of multiplex and real‐time PCR (Guo et al., 2018, 2020; Iwobi et al., 2015; Köppel et al., 2013; Xu et al., 2018). In addition, the inclusion of endogenous control reflects authentically the normal amplification reaction and decreases the occurrence of false negative results (Bacich et al., 2011; Guo et al., 2018, 2020; Li et al., 2019).…”

“…Taq Man real‐time PCR, based on probes labeled by different fluorescent reporters, combines the advantages of multiplex and real‐time PCR (Guo et al., 2018, 2020; Iwobi et al., 2015; Köppel et al., 2013; Xu et al., 2018). In addition, the inclusion of endogenous control reflects authentically the normal amplification reaction and decreases the occurrence of false negative results (Bacich et al., 2011; Guo et al., 2018, 2020; Li et al., 2019). Such Taq Man‐based triplex real‐time PCR methods for poultry authentication including chicken, duck, goose, and endogenous control, albeit appropriate, have not been developed so far.…”