Detection of the thaumatin-like protein gene from Brassica rapa var. oleifera in honey with the Proofman-LMTIA method

Wei Yao ^[1] ; Dandan Xu ^[1] ; Dan Zhou ^[1] ; Jinxin Liu ^[2] ; Xiaohua Zhang ^[2] ; Yao Wang ^[1] ; Deguo Wang ^[2]
1. [1] Henan University of Science and Technology
  
  Henan University of Science and Technology
  
  China
2. [2] Xuchang University
  
  Xuchang University
  
  Jian'an District, China
Localización: CyTA: Journal of food, ISSN 1947-6337, ISSN-e 1947-6345, Vol. 22, Nº. 1, 2024
Idioma: inglés
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Resumen
- Honey adulteration has been on the rise with the increasing demand for high-quality honey. The ladders-shape melting temperature isothermal amplification (LMTIA) was a newly developed nucleic acid amplification technique, and the aim of this study was to establish a method combining the LMTIA with the proofreading enzyme-mediated probe cleavage (Proof-man probe) for detecting the thaumatin-like protein (TLP) genes of Brassica rapa var. oleifera in honey. The LMTIA primers was selected as a target to be designed, the temperature of the Proof-man LMTIA reaction was optimized, and the specificity, sensitivity, and applicability to the honey samples were determined. The results showed that the Proof-man LMTIA method was highly specific with a detection sensitivity of 30-40 copies genomic DNA/reaction at the optimal temperature of 60°C, and the amplification could be completed within 20 min. This method can detect the adulteration of low-price B. rapa honey into high-price honey.