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A Novel Quantification Method for Gene-Edited Animal Detection Based on ddPCR

文献类型： 外文期刊

作者： Wang, Kaili 1 ; Ji, Yi 2 ; Peng, Cheng 2 ; Wang, Xiaofu 2 ; Yang, Lei 2 ; Lan, Hangzhen 1 ; Xu, Junfeng 2 ; Chen, Xiaoyun 2 ;

作者机构： 1.Ningbo Univ, Sch Food Sci & Engn, Ningbo 215211, Peoples R China

2.Minist Agr & Rural Affairs, Key Lab Traceabil Agr Genetically Modified Organis, State Key Lab Managing Biot & Chem Threats Qual &, Hangzhou 310021, Peoples R China

3.Zhejiang Acad Agr Sci, Zhejiang Key Lab Crop Germplasm Innovat & Utilizat, Hangzhou 310021, Peoples R China

关键词： gene editing; MSTN; nucleic acid detection; ddPCR

期刊名称：BIOLOGY-BASEL （ 影响因子：3.5； 五年影响因子：4.0 ）

ISSN：

年卷期： 2025 年 14 卷 2 期

页码：

收录情况： SCI

摘要： As gene-editing technologies continue to evolve, gene-edited products are making significant strides. These products have already been commercialized in the United States and Japan, prompting global attention to their safety and regulatory oversight. However, the detection of gene editing still relies on qPCR, and there is a lack of quantitative detection methods to quantify gene-editing components in products. To ensure consumer safety and transparency, we developed a novel droplet digital PCR (ddPCR)-based detection method for gene-edited products. Primers and probes were designed targeting the editing sites of MSTN-edited cattle, and the method was evaluated for specificity, sensitivity, real sample testing, and detection thresholds. Our results demonstrate that this ddPCR method is highly specific, with a detection limit of 5 copies/mu L, and it successfully detected MSTN edits in all 11 tested samples. Tests using both actual gene-edited cattle samples and plasmid DNA at concentrations of 5%, 1%, and 0.01% yielded consistent results, indicating the method's suitability for real-world applications. This ddPCR assay provides a sensitive and specific tool for detecting MSTN gene-edited cattle and determining the presence of gene-edited products, offering crucial support for regulatory monitoring of gene-edited animal-derived foods.

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