PCR荧光探针法猪源性成分试剂盒质量评价方法建立及应用  

Establishment and application of quality evaluation method for fluorescent probe PCR-based porcine ingredients detection kit

作　　者：马温华 张娟[2] 王延滨[3] 刘浩田 刘静 田雪梅 陈飞秀 赵蕾 陈爱亮[2] MA Wenhua;ZHANG Juan;WANG Yanbin;LIU Haotian;LIU Jing;TIAN Xuemei;CHEN Feixiu;ZHAO Lei;CHEN Ailiang(Institute of Forensic Science,Ministry of Public Security(MPS)&MPS Key Laboratory of Forensic Genetics,Beijing,100038,China;Institute of Quality Standards&Testing Technology for Agro-Products,Chinese Academy of Agricultural Sciences;Xicheng Branch of Beijing Public Security Bureau)

机构地区：[1]公安部鉴定中心,法医遗传学公安部重点实验室,北京市现场物证检验工程技术研究中心,现场物证溯源技术国家工程实验室,北京100038 [2]中国农业科学院农业质量标准与检测技术研究所 [3]北京市公安局西城分局刑警支队

出　　处：《质量安全与检验检测》2025年第2期27-32,共6页QUALITY SAFETY INSPECTION AND TESTING

基　　金：国家重点研发计划项目(2023YFF0614600);中央级公益性科研院所基本科研业务费专项资金项目(2023JB025)。

摘　　要：为了评价市售猪源性PCR荧光探针法核酸检测试剂盒的质量,本研究对市售5个品牌猪源性PCR荧光探针法核酸检测试剂盒进行质量评价。按照5种品牌试剂盒内置的说明书进行样品前处理、实时荧光定量PCR扩增以及结果判定,对精密度、特异性、检测限、冻融稳定性和模拟掺假样品检测5个指标进行了评价,探讨该5种试剂盒的检测性能。结果显示,5种PCR荧光探针法猪源性成分检测试剂盒的变异系数均小于5%,DNA检测限均可达到0.1 ng/μL,冻融稳定性实验变异系数小于5%,扩增效率可达到90%~110%,Kit_1#、Kit_2#、Kit_3#、Kit_4#试剂盒除了猪源性成分有扩增产物外,鸡、鸭、牛、羊等有非特异扩增,Kit_5#试剂盒只有猪源性成分有扩增产物,其他物种无扩增产物。5种PCR荧光探针法猪源性成分检测试剂盒,其精密度、检测限、稳定性等性能指标,符合行业标准YY/T 1182—2020《核酸扩增检测用试剂(盒)》中的要求。Kit_1#、Kit_2#、Kit_3#、Kit_4#试剂盒的特异性较差,容易引起误判,不适于猪肉及其肉制品的掺假鉴定。Kit_5#试剂盒的特异性好,可用于肉类食品中猪源性成分的检验鉴定。In order to evaluate the quality of commercially available porcine PCR fluorescent probe nucleic acid detection kits,this study evaluated the quality of 5 brands of commercially available porcine PCR fluorescent probe nucleic acid detection kits.According to the instructions of 5 brands of kits,sample pretreatment,real-time fluorescent PCR amplification and result determination were carried out.5 indexes of precision,specificity,detection limit,freeze-thaw stability and detection of simulated adulterated samples were evaluated,and the detection performance of the 5 kits was discussed.The results showed that the coefficient of variation of 5 PCR fluorescent probe kits for porcine component detection was less than 5%,the detection limit of DNA was up to 0.1 ng/μL,the coefficient of variation of freezing-thawing stability test was less than 5%,and the amplification efficiency was up to 90%-110%.Chickens,ducks,cattle,sheep have non-specific amplification except pig in Kit_1#,Kit_2#,Kit_3#,Kit_4#.In Kit_5#,only the pig-derived components had amplified products,and no amplified products in other species.The precision,detection limit,stability and other performance indicators of the 5 PCR fluorescent probe detection kits for porcine components met the requirements of industry standard YY/T 1182-2020"Nucleic acid amplification test reagents(kits)".The specificity of Kit_1#,Kit_2#,Kit_3#and Kit_4#is poor,which is easy to cause misjudgment,and is not suitable for the adulteration identification of pork and meat products.Kit_5#has good specificity and can be used for the detection and identification of porcine components in meat products.

关 键 词：猪源性成分检测 试剂盒 Q-PCR 质量评价 

分 类 号：TS251.7[轻工技术与工程—农产品加工及贮藏工程]