Naked Eye Meets Nucleic Acids: A Review on Dyes for Isothermal Amplification Methods for Detection of Meat Adulteration.

Meat adulteration poses significant challenges to public health and consumer trust, necessitating the development of effective detection methods. Isothermal nucleic acid amplification (NAA) techniques, such as loop-mediated isothermal amplification (LAMP) and polymerase spiral reaction (PSR), have emerged as rapid and cost-effective alternatives to conventional polymerase chain reaction (PCR). These techniques simplify infrastructure requirements and can enable naked-eye detection (NED) through visible color changes with specific dyes. This study highlights the potential and suitability of various dyes for the NED of IA-based meat adulteration detection. Intercalating dyes, such as SYBR Green I, bind to the amplified DNA and emit fluorescence. pH-sensitive dyes, including phenol red, neutral red, and xylenol orange, change color with pH shifts during amplification. Triphenylmethane dyes, such as crystal violet and malachite green, directly interact with DNA, showing no pH dependence. Intercalating dyes, such as SYBR Green I, achieve superior sensitivity, often reaching 10 fg of target DNA. Conversely, other dyes like Hydroxynaphthol blue and other pH-sensitive and pH-independent dyes provide slightly lesser sensitivity, typically ranging from 100 fg to 1 pg. Dye-based NED combined with IA offers advantages such as rapid results, high sensitivity and specificity, suitability for field testing, and potential integration into lab-on-chip systems. However, further research is required to optimize dye formulations, develop multiplex assays, enhance sample preparation for complex food matrices, and investigate novel isothermal methods and primer designs. Accurate standardization and validation of these techniques are crucial for their widespread adoption to ensure food safety and consumer trust in the meat industry.