Abstract
Milk ingredients are often included in a wide range of meat products, such as cooked hams and sausages, to improve technological characteristics. However, milk proteins are also important food allergens. The aim of this study was the development of a highly sensitive and specific real-time PCR system targeting the 12S rRNA gene of Bos domesticus for the detection and quantification of milk as an allergenic ingredient in processed meat products. The method was able to achieve an absolute limit of detection (LOD) of 6 fg of milk DNA. Using a normalized approach (∆Ct method) for the detection of milk protein concentrate (MPC), it was possible to obtain sensitivities down to 0.01% (w/w) of MPC in model hams (raw and cooked) and autoclaved sausages, and 0.005% in raw sausage mixtures. The developed systems generally presented acceptable PCR performance parameters, being successfully validated with blind samples, applied to commercial samples, and further compared with an immunochemical assay. Trace amounts of milk material were quantified in two out of 13 samples, but the results mostly infer the excessive practice of the precautionary labeling.
Highlights
The meat industry uses a wide range of ingredients with specific technological properties, to improve the appearance, taste and texture of products, as well as their nutritional value
In terms of relative limit of detection (LOD) and polymerase chain reaction (PCR) efficiency, the results suggest that thermal treatment had no effect on the detection of milk protein concentrate (MPC) since the same LOD of 0.01% (100 mg/kg) was reached in both ham systems with PCR efficiencies of 102.4% and 94.9% for raw and cooked mixtures, respectively (Figure 3a, Table 2)
We propose the development of four normalized real-time PCR systems targeting the 12S rRNA mitochondrial gene of Bos domesticus to detect and quantify trace amounts of milk ingredients in complex meat products
Summary
The meat industry uses a wide range of ingredients with specific technological properties, to improve the appearance, taste and texture of products, as well as their nutritional value. The introduction of these ingredients is intended to decrease the production costs or is unintentional introduced due to bad manufacturing practices. Some of these ingredients are a source of major food allergens, such as milk, legumes, egg, celery or cereal gluten proteins, causing a high health risk to consumers suffering from food allergies [1]. Milk proteins are often added to meat products in order to improve their juiciness, texture, and flavor These proteins can be caseins or whey proteins, such as ß-lactoglobulin and α-lactalbumin, which are considered major food allergens. To guarantee the safety of 95% of milk-allergic patients, a threshold of clinical reactivity to milk of 30 mg/kg was established by Morisset et al [4], based on the consumption of 100 g of product
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