Current Biotechnology ›› 2026, Vol. 16 ›› Issue (4): 744-755.DOI: 10.19586/j.2095-2341.2026.0130
• Special Forum on Detection Technology for Genetically Modified Organisms • Previous Articles Next Articles
Hua ZHANG1(
), Yuteng ZHANG2,3, Xinyao PEI1, Litao YANG2,3(
)
Received:2026-06-01
Accepted:2026-07-31
Online:2026-07-25
Published:2026-09-11
Contact:
Litao YANG
CLC Number:
Hua ZHANG, Yuteng ZHANG, Xinyao PEI, Litao YANG. Research Progress on Detection Methods of Genetically Modified and Gene-edited Products[J]. Current Biotechnology, 2026, 16(4): 744-755.
| 技术类别 | 代表方法 | 主要优势 | 主要局限性 | 适用场景 |
|---|---|---|---|---|
| 普通PCR | 元件特异性PCR、构建特异性PCR、事件特异性PCR、多重PCR | 操作相对简单、成本较低,适合已知靶标的快速初筛和基础鉴定 | 以定性或半定量分析为主,对低拷贝靶标和复杂基质样品的定量能力有限 | 转基因元件筛查、事件初步确认、未授权成分定性筛查 |
| qPCR | TaqMan探针qPCR、双重qPCR、多重qPCR | 灵敏度和特异性较高,定量能力成熟,标准化程度高,是监管检测中的核心方法 | 依赖标准曲线和参考物质;对单碱基编辑、小片段变异等靶标的识别能力受引物探针设计影响较大 | 转基因成分定量分析、事件特异性检测、监管阈值判定 |
| dPCR | ddPCR、芯片数字PCR、腔室数字PCR | 可实现绝对定量,无需标准曲线,适用于低拷贝、低丰度和复杂基质样品 | 仪器和耗材成本较高,通量和多重检测能力受平台限制,跨平台标准化仍需完善 | 低拷贝靶标定量、标准物质定值、基因编辑缺失位点精准识别 |
| 等温扩增 | LAMP、RPA、ERA等 | 反应条件简单、扩增速度快,对仪器依赖较低,便于现场快速检测 | 非特异性扩增和污染风险相对较高,定量能力和结果标准化程度通常弱于qPCR/dPCR | 现场筛查、口岸快速检测、基层实验室快速预警 |
| CRISPR/Cas辅助检测 | LAMP-CRISPR/Cas、RPA-CRISPR/Cas、ERA-CRISPR/Cas、多重CRISPR检测 | 兼具扩增快速性和CRISPR/Cas识别特异性,可与荧光、侧向层析和手机判读结合 | 体系设计较复杂,对crRNA、PAM、酶活性和反应条件依赖较强,多靶标定量和标准化验证仍需加强 | 转基因快速筛查、多靶标联检、现场可视化检测 |
| 免疫检测与生物传感 | 免疫层析试纸条、荧光侧向流免疫分析、电化学基因传感器、光谱融合检测 | 检测速度快、操作便捷,可实现可视化、便携化或无损识别,适合现场应用 | 多数方法依赖高质量抗体、探针或传感界面,对复杂基质干扰和结果定量稳定性要求较高 | 性状蛋白快速筛查、加工样品初筛、现场判读和快速分流 |
| 高通量测序与生物信息学分析 | 靶向测序、低深度NGS、长读长测序、RNA-seq、双链测序、遗传指纹分析 | 可同时获得插入位点、侧翼序列、突变类型和遗传背景信息,分辨率高,适合未知样品和基因编辑产品分析 | 成本、数据分析和数据库依赖较高,对结果解释、溯源判定和监管标准化提出更高要求 | 未知转基因事件识别、插入结构解析、基因编辑产品溯源和多位点分子指纹构建 |
Table 1 Methodological comparison of main detection techniques for genetically modified and gene edited products
| 技术类别 | 代表方法 | 主要优势 | 主要局限性 | 适用场景 |
|---|---|---|---|---|
| 普通PCR | 元件特异性PCR、构建特异性PCR、事件特异性PCR、多重PCR | 操作相对简单、成本较低,适合已知靶标的快速初筛和基础鉴定 | 以定性或半定量分析为主,对低拷贝靶标和复杂基质样品的定量能力有限 | 转基因元件筛查、事件初步确认、未授权成分定性筛查 |
| qPCR | TaqMan探针qPCR、双重qPCR、多重qPCR | 灵敏度和特异性较高,定量能力成熟,标准化程度高,是监管检测中的核心方法 | 依赖标准曲线和参考物质;对单碱基编辑、小片段变异等靶标的识别能力受引物探针设计影响较大 | 转基因成分定量分析、事件特异性检测、监管阈值判定 |
| dPCR | ddPCR、芯片数字PCR、腔室数字PCR | 可实现绝对定量,无需标准曲线,适用于低拷贝、低丰度和复杂基质样品 | 仪器和耗材成本较高,通量和多重检测能力受平台限制,跨平台标准化仍需完善 | 低拷贝靶标定量、标准物质定值、基因编辑缺失位点精准识别 |
| 等温扩增 | LAMP、RPA、ERA等 | 反应条件简单、扩增速度快,对仪器依赖较低,便于现场快速检测 | 非特异性扩增和污染风险相对较高,定量能力和结果标准化程度通常弱于qPCR/dPCR | 现场筛查、口岸快速检测、基层实验室快速预警 |
| CRISPR/Cas辅助检测 | LAMP-CRISPR/Cas、RPA-CRISPR/Cas、ERA-CRISPR/Cas、多重CRISPR检测 | 兼具扩增快速性和CRISPR/Cas识别特异性,可与荧光、侧向层析和手机判读结合 | 体系设计较复杂,对crRNA、PAM、酶活性和反应条件依赖较强,多靶标定量和标准化验证仍需加强 | 转基因快速筛查、多靶标联检、现场可视化检测 |
| 免疫检测与生物传感 | 免疫层析试纸条、荧光侧向流免疫分析、电化学基因传感器、光谱融合检测 | 检测速度快、操作便捷,可实现可视化、便携化或无损识别,适合现场应用 | 多数方法依赖高质量抗体、探针或传感界面,对复杂基质干扰和结果定量稳定性要求较高 | 性状蛋白快速筛查、加工样品初筛、现场判读和快速分流 |
| 高通量测序与生物信息学分析 | 靶向测序、低深度NGS、长读长测序、RNA-seq、双链测序、遗传指纹分析 | 可同时获得插入位点、侧翼序列、突变类型和遗传背景信息,分辨率高,适合未知样品和基因编辑产品分析 | 成本、数据分析和数据库依赖较高,对结果解释、溯源判定和监管标准化提出更高要求 | 未知转基因事件识别、插入结构解析、基因编辑产品溯源和多位点分子指纹构建 |
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