食品中马源性成分定性检测能力验证结果与分析

Validation results and analysis for qualitative detection of horse-derive components in food

中文关键词:  马源性成分  实时荧光PCR法  定性检测  能力验证

英文关键词:horse derived materials  real-time fluorescence PCR  qualitative detection  capability verification

中文摘要:

      目的 提高实验室对食品中马源性成分定性鉴定的准确性和评估检测人员的专业技术水平, 增强实验室竞争力。方法 通过核酸蛋白仪器分析法和单重实时荧光PCR法得到样品真核生物18S rRNA基因扩增的循环阈值, 进而分析DNA的提取质量。再利用标准SN/T 3730.5-2013《食品及饲料中常见畜类品种的鉴定方法第5部分: 马成分检测实时荧光PCR法》, 对该标准扩增体系的引物探针浓度进行优化和检出限分析后, 用优化的扩增体系对样品进行检测。再依据标准SB/T 10923-2012《肉与肉制品中动物源性成分的测定实时荧光PCR法》的要求, 对样品进行检测。最后对2个标准的检测结果进行比对。结果 提取的DNA质量符合标准要求。标准SN/T 3730.5-2013的引物探针浓度优化后, 能有效扩增阳性样品, 检出限为0.1%, 能有效检测马源性成分。2个标准检测20-M805和20-Q719待测样品, 均未检出马源性成分。结论 本次能力验证获得满意评价。DNA提取质量的评估, 反应体系中引物探针浓度的优化与选择, 不同检测方法结果的相互验证和实验的质量控制都是影响能力验证结果的重要因素。

英文摘要:

      Objective To improve the accuracy of qualitative detection ability of horse-derive components in food and the professional technical level of the detection personnel, and enhance the competitiveness of the laboratory. Methods The DNA extraction quality was analyzed by cyclic threshold for amplification of eukaryotic gene 18S rRNA obtained by nucleic acid protein instrumental analysis and simplex real-time fluorescence PCR. After optimizing the primers and probe concentration and analyzing the detection limit of the amplification system in SN/T3730.5-2013 Identification of domestic animal ingredient in food and feed-Part5: Detection of horse ingredient-Real-time PCR method, the optimized amplification system was used to detected the samples. The samples were also detected according to the requirements of SB/T 10923-2012 Identification of animal derived materials in meat and meat products. Finally, the results of the 2 standards were compared. Results The quality of the extracted DNA met the standard requirements. The optimized primer probe concentration of SN/T 3730.5-2013 could effectively amplify positive samples. The detection limit was 0.1%. It can effectively detect horse derived components. The horse derived components were not detected in 20-M805 and 20-Q719 samples by two methods. Conclusion The ability verification is satisfied. Evaluation of DNA extraction quality, optimization and selection of primer and probe concentration in reaction system, mutual verification of results of different detection methods and quality control of experiments are all important factors affecting the results of proficiency test.