食品检测用单核细胞增生李斯特氏菌标准物质的研制
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摘要
目的建立食品检测用单核细胞增生李斯特氏菌标准物质的制备方法,研制均匀稳定的标准物质。方法利用冷冻干燥法制备103 CFU/样品的标准物质,参照CNAS-GL 29《标准物质/标准样品定值的一般原则和统计方法》,抽取20件样品,利用平板计数法测定标准物质的均匀性,并对结果进行统计分析;将样品分别置于-20、25、37℃条件下保藏,对其储藏稳定性和运输稳定性进行评价。组织5家实验室进行协同标定,使用30种即食熟肉制品作为基质,并按照国标法检验标物物质的适用性。结果采用单因素方差分析进行均匀性检验, F=0.922,符合标准物质的要求。标准物质在-20℃保藏28 d, 25、37℃保藏7 d,样品仍然稳定。经5家实验室协同标定,样品含量均在103 CFU/样品的水平;标准物质加入到30种即食熟肉制品中,均可以检出单核细胞增生李斯特氏菌。结论本研究所制备的单核细胞增生李斯特氏菌标准物质的均匀性、储藏稳定性和运输稳定性均符合要求,适用性良好,可用于食品检测实验室的质量控制和食品中单核细胞增生李斯特氏菌检测结果的评价。
Objective To establish the method for preparing Listeria monocytogenes reference material for food analysis,and develop homogenous and stable reference material.Methods The standard substance of 103 CFU/sample was prepared by freeze-drying method.According to CNAS-GL 29 General principles and statistical methods for the determination of standard substances/standard samples,20 samples were selected,the homogeneity of standard substance was determined by plate counting method and the results were analyzed statistically.The samples were stored at-20,25,and 37 °C,respectively,and their storage stability and transport stability were evaluated,then 5 laboratories were organized for collaborative calibration,30 ready-to-eat cooked meat products were used as the matrix,and the applicability of the target substance was tested according to the national standard method.Results The homogeneity was analyzed by one-way anova,F=0.922,which indicated the reference material prepared in the study could meet the requirement of homogeneity for reference material.The reference material remained stable after storage at-20 ℃ for 28 d,25 ℃ and 37 ℃ for 14 d.Additionally,the sample was confirmed at the level of 103 CFU/sample by the collaborative calibration with 5 laboratories.Added to 30 kinds of ready-to-eat meat products,the Listeria monocytogenes in the reference material remained detectable.Conclusion The homogeneity,storage stability and transportation stability of Listeria monocytogenes reference material prepared in this study all meet the requirements and have good applicability,which can be used for quality control of food testing laboratory and evaluation of detection results of Listeria monocytogenes in food.
Objective To establish the method for preparing Listeria monocytogenes reference material for food analysis,and develop homogenous and stable reference material.Methods The standard substance of 103 CFU/sample was prepared by freeze-drying method.According to CNAS-GL 29 General principles and statistical methods for the determination of standard substances/standard samples,20 samples were selected,the homogeneity of standard substance was determined by plate counting method and the results were analyzed statistically.The samples were stored at-20,25,and 37 °C,respectively,and their storage stability and transport stability were evaluated,then 5 laboratories were organized for collaborative calibration,30 ready-to-eat cooked meat products were used as the matrix,and the applicability of the target substance was tested according to the national standard method.Results The homogeneity was analyzed by one-way anova,F=0.922,which indicated the reference material prepared in the study could meet the requirement of homogeneity for reference material.The reference material remained stable after storage at-20 ℃ for 28 d,25 ℃ and 37 ℃ for 14 d.Additionally,the sample was confirmed at the level of 103 CFU/sample by the collaborative calibration with 5 laboratories.Added to 30 kinds of ready-to-eat meat products,the Listeria monocytogenes in the reference material remained detectable.Conclusion The homogeneity,storage stability and transportation stability of Listeria monocytogenes reference material prepared in this study all meet the requirements and have good applicability,which can be used for quality control of food testing laboratory and evaluation of detection results of Listeria monocytogenes in food.
引文
[1]Jordan K,Mcauliffe O.Listeria monocytogenes in foods[J].Adv Food Nutr Res,2018,(86):181-213.
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[4]GB 4789.30-2016食品安全国家标准食品微生物学检验单核细胞增生李斯特氏菌检验.GB 4789.30-2016 National food safety standard-Food microbiological examination-Listeria monocytogenes[S].
[5]胡小玲,杨更发.微生物检测质量控制标样在微生物实验室内质量控制中的应用[J].中国卫生检验杂志,2007,17(12):2320-2321.Hu XL,Yang GF.The application of microbiological detecting reference material in internal quality control of microbiological laboratories[J].Chin J Health Lab Technol,2007,17(12):2320-2321.
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[7]曹丽梅.我国生物检测用国家标准物质现状与思考[J].中国生物制品学杂志,2015,28(8):886-888.Cao LM.Current situation and thinking of national standard substance for biological testing in China[J].Chin J Biol,2015,28(8):886-888.
[8]吴谦,黄晓蓉,王晶,等.鸡肉中单核细胞增生李斯特氏菌标准物质的研制[J].计量学报,2010,31(z1):9-13.Wu Q,Huang XR,Wang J,et al.The preparation of reference materials of Listeria monocytogenes in the matrix of chicken[J].Acta Metrol Sin,2010,31(z1):9-13.
[9]薛蕾,隋志伟,张玲,等.金黄色葡萄球菌标准物质的研制[J].食品科学,2015,36(8):44-48.Xu L,Sui ZW,Zhang L,et al.Preparation of Straphylococcus aureus reference material[J].Food Sci,2015,36(8):44-48.
[10]CNAS-GL 03能力验证样品均匀性和稳定性评价指南[S].CNAS-GL 03 2006 Guidance on evaluating the homogeity and stablity of samples used for proficiecy testing[S].
[11]CNAS-GL 29标准物质/标准物质(样品)定值的一般原则和统计方法[S].CNAS-GL 29 Reference materials-General and statistical priciples for certification[S].
[12]叶子弘,赵煦泓,邹克琴,等.生物标准物质研究简析[J].中国计量,2009,(4):67-69.Ye ZH,Zhao XH,Zou KQ,et al.The analysis of biological reference material study[J].China Metrol,2009,(4):67-69.
[13]张洁.生物标准物质的制备、发展及管理[J].中国生物制品学杂志,2007,20(8):630-632.Zhang J.The preparation,development and administration of biological reference material[J].Chin J Biol,2007,20(8):630-632.
[14]陈彬,郑晶,黄晓蓉,等.乳粉中阪崎肠杆菌标准物质的研制[J].中国乳品工业,2012,40(12):16-18.Chen B,Zheng J,Huang XR,et al.Preparation and certification of reference material for Enterobacter sakazakii in milk powder[J].China Dairy Ind,2012,40(12):16-18.
[15]柯璐,戴晓丽,林杰,等.志贺氏菌标准物质的制备方法[J].食品科学,2015,36(24):253-259.Ke L,Dai XL,Lin J,et al.The preparation of Shigella reference material[J].Food Sci,2015,36(24):253-259.
[16]柯璐,林杰,戴晓丽,等.肠炎沙门氏菌活菌标准物质的研制[J].食品研究与开发,2015,(5):111-115.Ke L,Lin J,Dai XL,et al.The reference materials of preparation about Salmonella enteritidis[J].Food Res Dev,2015,(5):111-115.
[17]薛蕾,林婧,隋志伟,等.大肠杆菌定量检测用标准物质的研制[J].计量学报,2015,36(6):652-656.Xue L,Lin J,Sui ZW,et al.Preparation of reference material for quantitative detection of Escherial coli[J].Acta Metrol Sin,2015,36(6):652-656.
[18]陈敏璠,傅博强,林婧,等.阴沟肠杆菌定量标准物质的研制[J].计量学报,2017,38(5):527-531.Chen MF,Fu BQ,Lin J,et al.Development of quantitative reference material of Enterobacter cloacae[J].Acta Metrol Sin,2017,38(5):527-531.
[2]Noordhout CM,Devleesschauwer B,Angulo FJ,et al.The global burden of Listeriosis:A systematic review and meta-analysis[J].Lancet Infect Dis,2014,14(11):1073-1082.
[3]GB 29921-2013食品安全国家标准食品中致病菌限量.GB 29921-2013 National food safety standard-Limit of pathogenic bacterial in food[S].
[4]GB 4789.30-2016食品安全国家标准食品微生物学检验单核细胞增生李斯特氏菌检验.GB 4789.30-2016 National food safety standard-Food microbiological examination-Listeria monocytogenes[S].
[5]胡小玲,杨更发.微生物检测质量控制标样在微生物实验室内质量控制中的应用[J].中国卫生检验杂志,2007,17(12):2320-2321.Hu XL,Yang GF.The application of microbiological detecting reference material in internal quality control of microbiological laboratories[J].Chin J Health Lab Technol,2007,17(12):2320-2321.
[6]丁丽霞,周海钧.生物标准物质的研究和技术要求[J].中国药师,2007,10(3):229-232.Ding LX,Zhou HJ.The study and technological requirements of biological reference materials[J].China Pharm,2007,10(3):229-232.
[7]曹丽梅.我国生物检测用国家标准物质现状与思考[J].中国生物制品学杂志,2015,28(8):886-888.Cao LM.Current situation and thinking of national standard substance for biological testing in China[J].Chin J Biol,2015,28(8):886-888.
[8]吴谦,黄晓蓉,王晶,等.鸡肉中单核细胞增生李斯特氏菌标准物质的研制[J].计量学报,2010,31(z1):9-13.Wu Q,Huang XR,Wang J,et al.The preparation of reference materials of Listeria monocytogenes in the matrix of chicken[J].Acta Metrol Sin,2010,31(z1):9-13.
[9]薛蕾,隋志伟,张玲,等.金黄色葡萄球菌标准物质的研制[J].食品科学,2015,36(8):44-48.Xu L,Sui ZW,Zhang L,et al.Preparation of Straphylococcus aureus reference material[J].Food Sci,2015,36(8):44-48.
[10]CNAS-GL 03能力验证样品均匀性和稳定性评价指南[S].CNAS-GL 03 2006 Guidance on evaluating the homogeity and stablity of samples used for proficiecy testing[S].
[11]CNAS-GL 29标准物质/标准物质(样品)定值的一般原则和统计方法[S].CNAS-GL 29 Reference materials-General and statistical priciples for certification[S].
[12]叶子弘,赵煦泓,邹克琴,等.生物标准物质研究简析[J].中国计量,2009,(4):67-69.Ye ZH,Zhao XH,Zou KQ,et al.The analysis of biological reference material study[J].China Metrol,2009,(4):67-69.
[13]张洁.生物标准物质的制备、发展及管理[J].中国生物制品学杂志,2007,20(8):630-632.Zhang J.The preparation,development and administration of biological reference material[J].Chin J Biol,2007,20(8):630-632.
[14]陈彬,郑晶,黄晓蓉,等.乳粉中阪崎肠杆菌标准物质的研制[J].中国乳品工业,2012,40(12):16-18.Chen B,Zheng J,Huang XR,et al.Preparation and certification of reference material for Enterobacter sakazakii in milk powder[J].China Dairy Ind,2012,40(12):16-18.
[15]柯璐,戴晓丽,林杰,等.志贺氏菌标准物质的制备方法[J].食品科学,2015,36(24):253-259.Ke L,Dai XL,Lin J,et al.The preparation of Shigella reference material[J].Food Sci,2015,36(24):253-259.
[16]柯璐,林杰,戴晓丽,等.肠炎沙门氏菌活菌标准物质的研制[J].食品研究与开发,2015,(5):111-115.Ke L,Lin J,Dai XL,et al.The reference materials of preparation about Salmonella enteritidis[J].Food Res Dev,2015,(5):111-115.
[17]薛蕾,林婧,隋志伟,等.大肠杆菌定量检测用标准物质的研制[J].计量学报,2015,36(6):652-656.Xue L,Lin J,Sui ZW,et al.Preparation of reference material for quantitative detection of Escherial coli[J].Acta Metrol Sin,2015,36(6):652-656.
[18]陈敏璠,傅博强,林婧,等.阴沟肠杆菌定量标准物质的研制[J].计量学报,2017,38(5):527-531.Chen MF,Fu BQ,Lin J,et al.Development of quantitative reference material of Enterobacter cloacae[J].Acta Metrol Sin,2017,38(5):527-531.