基于微滴数字PCR技术检测致敏原芒果成分
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(1. 北京市食品检验研究院 (北京市食品安全监控和风险评估中心 ) ,北京 100094 ; 2. 国家市场监管重点实验室 (食品安全重大综合保障关键技术 ) ,北京 100094)

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通讯作者:

李恩静(1983—),女,北京市食品检验研究院工程师,硕士。E-mail:enjingli@126.com

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国家重点研发计划项目(编号:2019YFC1605001)


Detection of allergen mango components by using droplet digital PCR technology
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(1. Beijing Institute of Food Inspection and Research (Beijing Municipal Center for Food Safety Monitoring and Risk Assessment , Beijing 100094 , China ; 2. Laboratory of Key Technologies of Major Comprehensive Guarantee of Food Safety for State Market Regulation , Beijing 100094 , China)

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    摘要:

    [目的]建立微滴数字 PCR 技术检测致敏原芒果成分的方法。[方法]选取叶绿体 trnH-psbA 基因作为致敏原芒果成分检测的靶基因,设计特异性引物探针,对微滴数字 PCR 方法反应条件进行优化,对方法的灵敏度和稳定性进行验证,并应用于 10批次食品样品中致敏原芒果成分检测。[结果]试验建立的微滴 PCR 方法特异性较好,不与芒果以外10种其他水果交叉反应,在上、下游引物和探针浓度分别为 200,200,100 nmol/L,退火温度为 52 ℃的最适微滴 PCR 反应条件下,方法的核酸水平检测灵敏度为 5 pg/μL,芒果质量含量检测灵敏度为 5 mg/kg,方法变异系数 <25%,重复性好,应用于 10批次食品样品检测结果与食品标签一致。[结论]该方法具有快速、特异性、灵敏度高的优点,适用于致敏原芒果成分的检测。

    Abstract:

    [[Objective ]] A droplet digital PCR method for detect allergen mango components was established in this study.[Methods ] Firstly,the chloroplast trnH-psbA gene was selected as the target gene for detect allergen mango components.Secondly,specific primers and probe were designed.Finally,the reaction conditions of droplet digital PCR was optimized to verify the sensitivity and stability of the method.[Results ] This method had been applied to detect allergen mango components in 10 batches of food samples.The results of establishment of droplet digital PCR method was showed good specificity and not cross -react with 10 kinds of other fruits except mango.Under the optimal ddPCR reaction conditions,which were to have the concentration of forward and reverse primers and probe set as 200,200 and 100 nmol/L,respectively,and to have annealing temperature set as 52 ℃,the detection sensitivity of nucleic acid level reached 5 pg/μL,and the detection sensitivity of mango content reached 5 mg/kg,the coefficient of variation of the method was less than 25%.The test results of 10 batches of food samples applied with the method were consistent with the food labels.[Conclusion ] This method has the advantages of being fast,specific,and highly sensitive,making it suitable for the detection of allergenic mango components.

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引用本文

李恩静,王 丹,李 龙,等.基于微滴数字PCR技术检测致敏原芒果成分[J].食品与机械,2024,40(9):50-55,115.
LI Enjing, WANG Dan, LI Long, et al. Detection of allergen mango components by using droplet digital PCR technology[J]. Food & Machinery,2024,40(9):50-55,115.

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  • 收稿日期:2023-11-14
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  • 在线发布日期: 2025-02-18
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