Detection of Honey Adulterated Rice Syrup by Real-time Fluorescence Spanning Rolling Circle Isothermal Amplification
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(1.College of Food Science and Technology, Hebei Agricultural University, Baoding 071000, Hebei;2.Department of Science and Engineering, Hebei Agricultural University, Cangzhou 061100, Hebei;3.College of Life Sciences, Hebei Agricultural University, Baoding 071000, Hebei;4.Key Laboratory of Analysis and Prevention and Control of Pathogenic Microorganisms Caused by Zootopia in Hebei Province, Baoding 071000, Hebei;5.School of Public Health, Hebei University, Baoding 071000, Hebei)

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    Abstract:

    In this study, It combined saltatory rolling circle amplification (SRCA) with fluorescence technology to establish a real-time fluorescence SRCA technique for detection of honey adulterated rice syrup. Three DNA extraction protocols were evaluated, four specific genes of rice (PDL, SPS, rbcL, GOS9) were used as target sequences to design primers, suitable primers were screened, amplification reaction conditions were optimized, and a method for detection of honey adulterated rice syrup was established in combination with fluorescence technology. The results showed that the sensitivity of the established real-time fluorescence SRCA method for the detection of rice DNA was 8.45×101 fg/μL, and its good specificity was confirmed by the specificity evaluation. The linear relationship between the logarithm of the proportion of adulteration and the Ct value was established in the manual simulated adulteration detection test, and the linear equation was y=6.618x+7.651 (R2=0.993), which can accurately detect the components of rice syrup in honey down to 1%. The method is sensitive, with low detection limit, and can quickly and accurately quantify the adulteration of honey with rice syrup, which provides a new idea for the rapid quantitative detection of honey adulteration.

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  • Received:January 30,2023
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  • Online: February 19,2024
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