Liu K et al. (2022), An Accurate and Rapid Way for Identifying Food ...

ECB-ART-51072

Foods 2022 Dec 21;121:. doi: 10.3390/foods12010017.

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An Accurate and Rapid Way for Identifying Food Geographical Origin and Authenticity: Editable DNA-Traceable Barcode.

Liu K , Xing R , Sun R , Ge Y , Chen Y .

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DNA offers significant advantages in information density, durability, and replication efficiency compared with information labeling solutions using electronic, magnetic, or optical devices. Synthetic DNA containing specific information via gene editing techniques is a promising identifying approach. We developed a new traceability approach to convert traditional digitized information into DNA sequence information. We used encapsulation to make it stable for storage and to enable reading and detection by DNA sequencing and PCR-capillary electrophoresis (PCR-CE). The synthesized fragment consisted of a short fragment of the mitochondrial cytochrome oxidase subunit I (COI) gene from the Holothuria fuscogilva (ID: LC593268.1), inserted geographical origin information (18 bp), and authenticity information from Citrus sinensis (20 bp). The obtained DNA-traceable barcodes were cloned into vector PMD19-T. Sanger sequencing of the DNA-traceable barcode vector was 100% accurate and provided a complete readout of the traceability information. Using selected recognition primers CAI-B, DNA-traceable barcodes were identified rapidly by PCR amplification. We encapsulated the DNA-traceable barcodes into amorphous silica spheres and improved the encapsulation procedure to ensure the durability of the DNA-traceable barcodes. To demonstrate the applicability of DNA-traceable barcodes as product labels, we selected Citrus sinensis as an example. We found that the recovered and purified DNA-traceable barcode can be analyzed by standard techniques (PCR-CE for DNA-traceable barcode identification and DNA sequencing for readout). This study provides an accurate and rapid approach to identifying and certifying products' authenticity and traceability.

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References [+] :

Antkowiak, Integrating DNA Encapsulates and Digital Microfluidics for Automated Data Storage in DNA. 2022, Pubmed

	Figure 1. (a) Overview of storing and retrieving data into and from DNA-traceable barcode vectors; (b) DNA-traceable barcodes synthesis schemes. Fragment 1 was 38 bp in length, and fragment 2 was 112 bp.
	Figure 2. The PCR product electrophoretogram. (a) The PCR product electrophoretogram of Approach 1. M: 700 bp DNA ladder; lane 1–4: blank; lane 5–8: the sample of the amplification of the DNA-traceable barcode; (b) The PCR product electrophoretogram of Approach 2. M: 700 bp DNA ladder; lane 1–4: blank; lane 5–8: the sample of the amplification of the DNA-traceable barcode.
	Figure 3. Detection of primers screening gel electrophoresis results. M: 700 bp DNA ladder; lane 1-4: blank; lane 5–8: the sample of the amplification of the CAI-A, CAI-B, CAI-C, CAI-D primers.
	Figure 4. SEM imaging of silica particles with encapsulated DNA-traceable barcode vector prepared by different protocols. (a) Blank: silica particles without encapsulated DNA-traceable barcode; 1–8: the nanoparticles prepared according to the synthesis scheme A–H in Table 2; (b) 1–4: the nanoparticles prepared according to the synthesis scheme G0–G3 in Table 2.
	Figure 5. Characterization and identification of DNA-traceable barcode encapsulated silica particles. (a) 1: Transmission electron microscopy image of SiO2 particles without encapsulated DNA-traceable barcode. 2: Transmission electron microscopy image of DNA/SiO2 particles. Scale bar, 50 nm. The silica layer has a thickness of 12 nm, and it protects the nucleic acid from ROS and heat. (b) Capillary electrophoresis graph obtained from plasmid DNA before encapsulation and after release from DNA/SiO2 particles. Purple lines are upper marker, green lines are lower marker.
	Figure 6. Recovery and detection of DNA-traceable barcode labels for Citrus sinensis.
	Figure 7. Capillary electrophoresis graph for DNA-traceable barcode labeling of Citrus sinensis. Purple lines are upper marker, green lines are lower marker. L is Ladder; lane 1–3 is the samples not labeled with DNA-traceable barcode; lane 4–12 is the samples labeled with DNA-traceable barcode.