Abstract
Deoxyribonucleic acid (DNA) extraction, which is the mainstay of molecular amplification for food pathogen detection procedure, is dependent on laboratory infrastructure, cumbersome methods, and time consumption. To tackle these dependencies, we developed an enhanced mussel-inspired cellulose card for the convenient, eco-friendly, and rapid DNA extraction from bacteria present in food. This enhanced cellulose mussel-inspired DNA adhesive card (BDA card) was successfully prepared using Na + through the self-polymerization of biopolymer polyvalent ions and 3,4-dihydroxy-l-phenylalanine (L-Dopa). We observed that, when the biopolymer surface modification of the cellulose was NaCl-doped, the DNA extraction and purification performances of the obtained BDA card were enhanced when compared to the performance of the cellulose modified with the biopolymer doped with HCl. After the surface modification, the previously modified cellulose card was characterized with the Fourier transform infrared spectroscopy (FTIR), X-ray diffraction (XRD), and scanning electron microscopy (SEM). The DNA extraction performances as well as the quality of the bacteria genomic DNA extracted were equally measured. The Na + -doped DNA adhesive cellulose card (BDA card) depicted the best bacteria genomic DNA extraction performances. When compared to commercially available bacteria genomic DNA extraction kits, the BDA card portrayed better DNA extraction performances, in a one-step extraction protocol. It allowed easy and convenient DNA extraction. The whole genomic DNA extraction process could be completed in 10 min. The concentration of the genomic DNA extracted from overnight bacteria culture was highest when extracted with the BDA card 459.39 ± 86 ng/µl as compared to Ezup and PureLink genomic DNA kits. When tested with spiked chicken, it was statistically equal to the Mag-MK food genomic DNA extraction kit. The proposed method provides a simple, efficient, eco-friendly, and convenient approach for bacteria genomic DNA extraction and purification from samples.
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Data Availability
The raw data used and analyzed in this research were generated at the Synergetic Innovation Center of Food Safety and Nutrition, State Key Laboratory of Food Science and Technology, School of Food Science. Derived data supporting the findings of this study are available from the corresponding author upon request.
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Funding
This study was funded by the 2020 Jiangsu Provincial Market Supervision Administration Science and Technology Plan Project (KJ204139), the National Key Research and Development Program of China 2018YFC1602800, the National Natural Science Foundation of China (31772069, 31801660), the Fundamental Research Funds for the Central Universities (1022050205205500), the national first-class discipline program of Food Science and Technology (JUFSTR20180303), and The Collaborative Innovation Center for Food Safety and Quality Control.
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Nanfack Vouffo Denise Carole declares that she has no conflict of interest. Jia Yang declares that he has no conflict of interest. Xiao Yuan declares that he has no conflict of interest. Jiadi Sun declares that she has no conflict of interest. Xiulan Sun declares that she has no conflict of interest. And Jian Ji declares that he has no conflict of interest.
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Carole D. V. Nanfack and Jia Yang are co-first authors.
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Nanfack, C.D.V., Yang, J., Yuan, X. et al. 3, 4-Dihydroxy-l-phenylalanine Biopolymer Cellulose DNA Adhesive Card as an Enhanced Solid-Phase One-Step DNA Extraction Method from Foodborne Pathogens in Food Samples. Food Anal. Methods 15, 2069–2083 (2022). https://doi.org/10.1007/s12161-021-02177-z
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DOI: https://doi.org/10.1007/s12161-021-02177-z