肉及肉制品牛源性成分核酸检测试剂盒评价技术的研究
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摘要
目的探索适合评价核酸检测试剂盒的评价方法。方法本研究采用室内技术参数验证的方法对某公司的肉及肉制品牛源性成分核酸检测试剂盒(实时荧光PCR法)进行评价。对检测的灵敏度特异性、假阳性/假阴性、检测限、耐受性及与标准方法的一致性共6个方面的参数分析评价。结果评价结果表明,对生鲜肉、酱卤肉、熏烤肉、速冻调理肉制品4类共40种肉制品,试剂盒检测结果与标准检测方法检测结果无显著差异,即试剂盒可以满足肉及肉制品牛源性成分核酸检测的需求。结论本文为检测机构筛选合适的牛源性成分检测试剂盒有一定的借鉴意义,并对建立核酸检测试剂盒的评价方法提供一定的技术帮助。
Objective To explore a suitable method for the evaluation of nucleic acid detection kit. Methods The meat and meat products nucleic acid detection kit(real-time PCR method) from biological composition was evaluated by the method of indoor technology parameter validation. Total of 6 parameters about the sensitivity, the specificity, and the false positives/false negatives, detection limit, tolerance and conformance with the standard method were analyzed and evaluated. Results There was no significant difference between the kit method and the reference method for 40 kinds of meat products about raw meat, pot-stewed meat, smoked meat, quick-frozen meat, which meant the kit method could meet our requirement for meet and meet product nucleic acid detection. Conclusion It has certain reference significance for screening institutions to screen suitable bovine source components detection kit, and provide some technical help for establishing the evaluation method of nucleic acid detection kit.
Objective To explore a suitable method for the evaluation of nucleic acid detection kit. Methods The meat and meat products nucleic acid detection kit(real-time PCR method) from biological composition was evaluated by the method of indoor technology parameter validation. Total of 6 parameters about the sensitivity, the specificity, and the false positives/false negatives, detection limit, tolerance and conformance with the standard method were analyzed and evaluated. Results There was no significant difference between the kit method and the reference method for 40 kinds of meat products about raw meat, pot-stewed meat, smoked meat, quick-frozen meat, which meant the kit method could meet our requirement for meet and meet product nucleic acid detection. Conclusion It has certain reference significance for screening institutions to screen suitable bovine source components detection kit, and provide some technical help for establishing the evaluation method of nucleic acid detection kit.
引文
[1]SB/T 10923-2012肉及肉制品中动物源性成分的测定实时荧光PCR法[S].SB/T 10923-2012 Identification of animal derived materials in meat and meat products-Real-time PCR methods[S]
[2]SN/T 2775-2011商品化食品检测试剂盒评价方法[S].SN/T 2775-2011 Methods for the evaluation of commercial test kits for food testing purpose[S].
[3]绍碧英,董建,陈彬,等.沙门氏菌核酸检测试剂盒的评价[J].食品科学,2015,24(36):242-245Shao BY,Dong J,Chen B,et al.Evaluation of nucleic acid detection kit of Salmonella[J].Food Sci,2015,24(36):242-245.
[4]董健,郑晶,林杰,等.3M PetrifilmTM金黄色葡萄球菌检验测试片的评价[J].食品与发酵科技,2015,50(4):56-60.Dong J,Zheng J,Lin J,et al.Validation of 3M PetrifilmTM Staphylococcus aures count plate[J].Food Ferment Technol,2015,50(4):56-60.
[5]黄晓蓉.食品检测试剂盒评价[J].检验检疫学报,2013,23(3):1-4.Huang XR.Validation of commercial food testing kit[J].China Inspect Quarant,2013,23(3):1-4.
[6]GB/T 6379.2-2004测量方法与结果的准确度(正确度与精密度)第2部分:确定标准测量方法重复性与再现性的基本方法[S]GB/T 6379.2-2004 Accuracy(trueness and precision)of measurement methods and results-Part 2:Basic method for the determination of repeatability and reproducibility of a standard measurement method[S].
[7]李慎安.JJF 1059-1999《测量不确定度评定与表示》讨论之四方法确认的重复性标准差与复现性标准差[J].工业计量,2006,(2):35-37.Li SA.The 4th of discussion about JJF 1059-1999 Measurement uncertainty evaluation and expression-Determine the repeatability of standard deviation and recurrence standard deviation[J].Ind Meas,2006,(2):35-37.
[8]GB/T 6379.4-2006/ISO5725-4:1994.测量方法与结果的准确度(准确度与精密度)第4部分:确定标准检测方法准确度的基本方法[S]GB/T 6379.4-2006/ISO5725-4:1994.Accuracy(trueness and precision)of measurement methods and results-Part 4:Bisic method for the determination of trueness of a standard measurement method[S]
[9]Zhang CL,Fowler MR,Scott NW,et al.A Taq man real-time PCR system for the identification and quantification of bovine DNA in meats,milks and cheeses[J].Food Control,2006,(9):1149-1158
[10]黄娅琳.高温烹饪对动物肌肉组织DNA降解的影响[J].四川动物,2012,(2):222-225Huang YL.Effect of high temperature cooking on DNA degradation in animal muscle tissue[J].Sichuan Anim,2012,(2):222-225.
[11]Nadia B,Sami F,Saloua S.Comparison of methods in the recovery and amplificability of DNA from fresh and processed sardine and anchovy muscle tissues[J].Food Chem,2011,129(2):665-671
[12]何建文,韩建林,罗玉柱.利用不同方法从深加工牦牛肉产品中提取基因组DNA效果的比较[J].生物技术通报,2010,(10):162-167.He JW,Han JL,Luo YZ.Comparison of equantity and quality of genomic DNAs extract from highly processed yak meat products using different methods[J].Biotechnol Bull,2010,(10):162-167.
[13]Wagner JK.Understanding FDA regulation of DTC genetic tests within the context of administrative law[J].Am J Human Genet,2010,87(4):451-456.
[14]International Organization for Standardization.ISO E.N.16140Microbiology of food and animal feeding stuffs-Protocol for the validation of alternative methods[S].
[15]刘维薇,郑特,徐科,等.国外实验室自建项目管理带给我们的启示[J].中华临床实验室管理电子杂志,2014,2(3):136-139Liu WW,Zheng T,Xu K,et al.Foreign laboratory self-built project management to bring us inspiration[J].Chin J Clin Lab Manag,2014,2(3):136-139.
[16]刘东来,石大伟,张春涛.美国对实验室研发诊断试剂的监管之路[J].检验检疫学报,2016,25(3):244-252.Liu DL,Shi DW,Zhang CT.Development of the United States regulatory policy about the laboratory development test[J].Chin J New Drugs,2016,25(3):244-252.
[2]SN/T 2775-2011商品化食品检测试剂盒评价方法[S].SN/T 2775-2011 Methods for the evaluation of commercial test kits for food testing purpose[S].
[3]绍碧英,董建,陈彬,等.沙门氏菌核酸检测试剂盒的评价[J].食品科学,2015,24(36):242-245Shao BY,Dong J,Chen B,et al.Evaluation of nucleic acid detection kit of Salmonella[J].Food Sci,2015,24(36):242-245.
[4]董健,郑晶,林杰,等.3M PetrifilmTM金黄色葡萄球菌检验测试片的评价[J].食品与发酵科技,2015,50(4):56-60.Dong J,Zheng J,Lin J,et al.Validation of 3M PetrifilmTM Staphylococcus aures count plate[J].Food Ferment Technol,2015,50(4):56-60.
[5]黄晓蓉.食品检测试剂盒评价[J].检验检疫学报,2013,23(3):1-4.Huang XR.Validation of commercial food testing kit[J].China Inspect Quarant,2013,23(3):1-4.
[6]GB/T 6379.2-2004测量方法与结果的准确度(正确度与精密度)第2部分:确定标准测量方法重复性与再现性的基本方法[S]GB/T 6379.2-2004 Accuracy(trueness and precision)of measurement methods and results-Part 2:Basic method for the determination of repeatability and reproducibility of a standard measurement method[S].
[7]李慎安.JJF 1059-1999《测量不确定度评定与表示》讨论之四方法确认的重复性标准差与复现性标准差[J].工业计量,2006,(2):35-37.Li SA.The 4th of discussion about JJF 1059-1999 Measurement uncertainty evaluation and expression-Determine the repeatability of standard deviation and recurrence standard deviation[J].Ind Meas,2006,(2):35-37.
[8]GB/T 6379.4-2006/ISO5725-4:1994.测量方法与结果的准确度(准确度与精密度)第4部分:确定标准检测方法准确度的基本方法[S]GB/T 6379.4-2006/ISO5725-4:1994.Accuracy(trueness and precision)of measurement methods and results-Part 4:Bisic method for the determination of trueness of a standard measurement method[S]
[9]Zhang CL,Fowler MR,Scott NW,et al.A Taq man real-time PCR system for the identification and quantification of bovine DNA in meats,milks and cheeses[J].Food Control,2006,(9):1149-1158
[10]黄娅琳.高温烹饪对动物肌肉组织DNA降解的影响[J].四川动物,2012,(2):222-225Huang YL.Effect of high temperature cooking on DNA degradation in animal muscle tissue[J].Sichuan Anim,2012,(2):222-225.
[11]Nadia B,Sami F,Saloua S.Comparison of methods in the recovery and amplificability of DNA from fresh and processed sardine and anchovy muscle tissues[J].Food Chem,2011,129(2):665-671
[12]何建文,韩建林,罗玉柱.利用不同方法从深加工牦牛肉产品中提取基因组DNA效果的比较[J].生物技术通报,2010,(10):162-167.He JW,Han JL,Luo YZ.Comparison of equantity and quality of genomic DNAs extract from highly processed yak meat products using different methods[J].Biotechnol Bull,2010,(10):162-167.
[13]Wagner JK.Understanding FDA regulation of DTC genetic tests within the context of administrative law[J].Am J Human Genet,2010,87(4):451-456.
[14]International Organization for Standardization.ISO E.N.16140Microbiology of food and animal feeding stuffs-Protocol for the validation of alternative methods[S].
[15]刘维薇,郑特,徐科,等.国外实验室自建项目管理带给我们的启示[J].中华临床实验室管理电子杂志,2014,2(3):136-139Liu WW,Zheng T,Xu K,et al.Foreign laboratory self-built project management to bring us inspiration[J].Chin J Clin Lab Manag,2014,2(3):136-139.
[16]刘东来,石大伟,张春涛.美国对实验室研发诊断试剂的监管之路[J].检验检疫学报,2016,25(3):244-252.Liu DL,Shi DW,Zhang CT.Development of the United States regulatory policy about the laboratory development test[J].Chin J New Drugs,2016,25(3):244-252.