基于改进的预扩增实时荧光定量聚合酶链式反应法检验虹鳟源性成分-食品安全质量检测学报-CSCIED科技核心评价数据库-手机版

基于改进的预扩增实时荧光定量聚合酶链式反应法检验虹鳟源性成分

Detection of Oncorhynchus mykiss source components based on improved pre-amplification real-time quantitative polymerase chain reaction method

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DOI	10.19812/j.cnki.jfsq11-5956/ts.20250415003
刊名	食品安全质量检测学报 Journal of Food Safety and Quality
年，卷(期)	2025, 16(13)
作者	白朵兰, 洪麟, 杨滴, 陈文超, 王佳琪, 方蕾, 刘彦泓, 郑秋月,
作者单位	大连民族大学生命科学学院; 生物技术与资源利用教育部重点实验室; 大连市检验检测认证技术服务中心; 国家市场监督管理总局技术创新中心(食用农产品安全快速检测与追溯)
摘要	目的建立改进的预扩增实时荧光定量聚合酶链式反应法(quantitative real-time polymerase chain reaction, qPCR)法检验虹鳟源性成分的方法。方法基于对现行标准中虹鳟qPCR方法进行验证和改进提高, 建立改进的预扩增qPCR方法, 并对两种qPCR方法进行比较。结果经验证, 在采用标准中qPCR方法对虹鳟的近种鱼种银鲑、大西洋鲑等鱼种进行特异性验证时有交叉反应, 基于设置预扩增反应步骤的改进qPCR方法特异性良好, 对于银鲑、大西洋鲑等近种鱼种未出现交叉扩增。qPCR和预扩增qPCR方法的扩增效率分别为90.25%、104.91%, 最低检出限分别为2.99×10–3 ng/μL (0.83~8.87 pg/μL, 95%置信区间)和1.50×10–4 ng/μL (0.09~0.27 pg/μL, 95%置信区间)。结论所改进的虹鳟源性成分预扩增qPCR方法具有高特异性、高灵敏度, 适用于食品样品中虹鳟源性成分的真实性和掺假检验。
Abstract	Objective To establish a pre-amplified quantitative real-time polymerase chain reaction (qPCR) method for the detection of Oncorhynchus mykiss source components. Methods The study developed an enhanced pre-amplified qPCR method, which was based on the validation and improvement of the existing standard qPCR method and compared the performance of both methods. Results Specificity testing of the standard qPCR method revealed cross-reactivity with closely related species, such as Coho salmon and Atlantic salmon. The established improved pre-amplified qPCR method, which incorporated a pre-amplification step, exhibited high specificity and no cross amplification was observed for Coho salmon, Atlantic salmon and other closely related species. The amplification efficiencies of the standard qPCR and the pre-amplified qPCR methods were 90.25% and 104.91%, respectively. The lowest limits of detection of the 2 kinds of methods were 2.99×10–3 ng/μL (0.83–8.87 pg/μL, 95% confidence interval) for the standard qPCR and 1.50×10–4 ng/μL (0.09–0.27 pg/μL, 95% confidence interval) for the pre-amplified qPCR. Conclusion The method established in this study has the characteristics of strong specificity, high sensitivity, and is suitable for authenticity and adulteration detection of Oncorhynchus mykiss source components in food products.
关键词	虹鳟源性成分;真实性检验;qPCR ;预扩增 qPCR ;
KeyWord	Oncorhynchus mykiss source; authenticity identification; quantitative real-time polymerase chain reaction; pre-amplified quantitative real-time polymerase chain reaction
基金项目
页码	18-25

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白朵兰, 洪麟, 杨滴, 陈文超, 王佳琪, 方蕾, 刘彦泓*, 郑秋月*. 基于改进的预扩增实时荧光定量聚合酶链式反应法检验虹鳟源性成分 [J]. 食品安全质量检测学报. 2025; 16; (13). 18 - 25.

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