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| 基于改进的预扩增qPCR建立虹鳟源性成分检验方法 |
| Establishment of detection method for Rainbow Trout(Oncorhynchus mykiss)source components by improved Pre-amplified qPCR |
| 投稿时间：2025-04-15 修订日期：2025-06-12 |
| DOI： |
| 中文关键词: 虹鳟源性成分 真实性检验 qPCR 预扩增 qPCR |
| 英文关键词:rainbow trout source(Oncorhynchus mykiss) Authenticity identification qPCR Pre amplified qPCR |
| 基金项目:国家市场监管总局科技计划项目(2023MK130) |
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| Author | Institution |
| BAi DuoLan | Dalian Minzu University |
| HONG Lin | Dalian Center for Certification and Food and Drug Control,Technology Innovation Center of Rapid Screening,State Administration for Market Regulation |
| YANG Di | Dalian Center for Certification and Food and Drug Control,Technology Innovation Center of Rapid Screening,State Administration for Market Regulation |
| CHEN Wenchao | Dalian Center for Certification and Food and Drug Control,Technology Innovation Center of Rapid Screening,State Administration for Market Regulation |
| WANG Jiaqi | Dalian Center for Certification and Food and Drug Control,Technology Innovation Center of Rapid Screening,State Administration for Market Regulation |
| LIU Yanhong | Dalian Center for Certification and Food and Drug Control,Technology Innovation Center of Rapid Screening,State Administration for Market Regulation |
| ZHENG Qiuyue | Key Laboratory of Biotechnology and Bioresources Utilization of Ministry of Education,College of Life Science,Dalian Minzu University |
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| 中文摘要: |
| 目的 建立改进的预扩增实时荧光定量PCR（Quantitative Real-time PCR）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% CI）和1.50×10-4 ng/μL （0.09~0.27 pg/μL，95% CI）。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。 |
| 英文摘要: |
| Objective To establish a pre-amplified qPCR method for the detection of rainbow trout-derived ingredients was established. 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: it produced amplification curves only for rainbow trout genomic DNA and none for closely related species, such as Coho salmon or Atlantic salmon.The amplification efficiencies of the standard qPCR and the pre-amplified qPCR methods were E = 90.25% and E = 104.91%, respectively. The lowest detection limits of the two methods were 2.99 × 10-3 ng/μL (0.83 ~ 8.87 pg/μL, 95% CI) for the standard qPCR and 1.50 × 10-4 ng/μL (0.09 ~ 0.27 pg/μL, 95% CI) 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 rainbow trout-derived ingredients in food products |
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