食品沙门氏菌现行国内外标准中选择性增菌和分离的等效性评估  

The equivalence trials of Salmonella selective enrichment and isolation procedures in standard methods worldwide

作　　者：刘玥[1,2] 刘夏 崔琳 陈敏 刘弘[1] 顾其芳[1] 胡屹[2,5] 许学斌[1] LIU Yue;LIU Xia;CUI Lin;CHEN Min;LIU Hong;GU Qifang;HU Yi;XU Xuebin(Shanghai Multiple Center for Disease Prevention and Control,Shanghai 200336,China;School of Public Health,Fudan University,Shanghai 200032 China;Technology Center of Animal Plant and Food Inspection and Quarantine of Shanghai Customs,Shanghai 201708,China;Shanghai Center for Clinical Laboratory,Shanghai 200126,China;China and Key Laboratory of Public Health,Fudan University,Shanghai 200032,China)

机构地区：[1]上海市疾病预防控制中心,上海200336 [2]复旦大学公共卫生学院,上海200032 [3]上海海关动植物与食品检验检疫技术中心,上海201708 [4]上海市临床检验中心,上海200126 [5]复旦大学公共卫生安全教育部重点实验室,上海200032

出　　处：《中国食品卫生杂志》2022年第3期474-481,共8页Chinese Journal of Food Hygiene

基　　金：国家重点研发计划(2017YFC1600104);国家科技重大专项(2018ZX10714002-003-009);上海市健康委青年课题(20174Y0141)。

摘　　要：目的评估5种现行国内与国际食品沙门氏菌标准中选择性增菌步骤的等效性。方法按照GB4789.28—2013的方法使用沙门氏菌参比菌株验证5种沙门氏菌选择性增菌液亚硒酸盐胱氨酸增菌液(SC)、四硫磺酸钠煌绿增菌液(TTB)、罗伯特增菌液(RV)、穆勒-考夫曼四硫酸盐新生霉素增菌液(MKTTn)、亚硒酸盐煌绿增菌液(SBG)和5种选择性琼脂平板沙门氏菌显色琼脂平板(CAS)、木糖-赖氨酸-脱氧胆酸盐琼脂平板(XLD)、木糖赖氨酸十四烷基硫酸钠琼脂平板(XLT4)、亚硫酸铋琼脂平板(BS)和赫氏肠道菌琼脂平板(HE)的生长率(PR)和特征性生长;按照ISO 16140—2:2016和传统统计学方法设计路径,经高污染样品检测比较5种选择性增菌液和2种选择性琼脂平板的组合方法的敏感性、相对真值和可接受限值,评估10种增菌分离组合的分离敏感性、特异性、阳性预测值和阴性预测值。结果SC对猪霍乱和猪霍乱沙门氏菌孔成道夫变种存在无效增殖;CAS和XLD识别伤寒和非伤寒、硫化氢差异表达沙门氏菌能力明显优于XLT4、HE、BS;84件样品(畜肉22件、禽肉32件和水产品30件)经5种增菌组合确认25件阳性(29.76%)、鉴定分型42株沙门氏菌。5种组合方法的敏感性分别为:84.00%、100.00%、88.00%、88.00%和92.00%,没有统计学差异(P>0.05)。按照ISO 16140—2:2016释义组合1(SC+TTB)因SC存在漏检导致可接受限值超限;10种增菌组合中以RV-CAS综合效益最好,敏感性、特异性、阳性预测值、阴性预测值分别为:68.00%、95.00%、85.00%、88.00%。结论SC因窄谱增菌效果,影响方法敏感性、导致负偏差结果偏倚,使可接受限值超限值;增菌组合推优RV-CAS,平板组合推荐CAS+XLD;建议GB4789.4能完善沙门氏菌血清鉴定,加强应对流通样品混合菌型污染及接轨国际食品微生物标准。Objective To evaluate the equivalence of 5 Salmonella selective enrichment procedures in standard methods worldwide.Methods The growth rates(PR)and the colony characteristics of 5 selective enrichment broths(SC,TTB,RV,MKTTn,SBG)and 5 selective mediums(CAS,XLD,XLT4,HE,BS)were validated according to the GB4789.28—2013.The sensitivity(SE),the relative trueness(RT)and the acceptable limit(AL)were estimated on highly contaminated foods through the use of 5 protocols combined with 5 selective enrichment broths and 2 selective mediums according to the method of ISO 16140—2:2016.The sensitivity(Se),the specificity(Sp),the positive predictive value(PPV)and the negative predictive value(NPV)were simultaneously determined through the use of 10compound modes combined with selective enrichment broths and selective mediums according to the classical statistical methods.ResultsThe results showed that SC broth had no efficacy in enriching Salmonella enterica serotype Cholerae and Salmonella enterica serotype Cholerae var.Kunzendorf.The CAS and XLD were significantly better than XLT4,HE and BS in identifying typhoidal Salmonella and nontyphoidal Salmonella as well as the phenotype of hydrogen sulfide.Totally 84 samples(22 pieces of meat,32 pieces of poultry and 30 pieces of aquatic products)were tested.25 out of the 84(29.76%)were confirmed positive by 5 protocols,and 16 serotypes from 42 isolates were identified.The SE of 5protocols were 84.00%,100.00%,88.00%,88.00%and 92.00%,respectively,but showed no statistically significant difference between each other.The AL of one protocol(SC+TTB)went beyond the limitation of the ISO 16140—2:2016due to the ineffectiveness of SC broth.The compound mode of RV-CAS had the best comprehensive benefits,while the Se,the Sp,the PPV and the NPV of which were 68.00%,95.00%,85.00%and 88.00%,respectively.Conclusion The ineffectiveness of SC broth reduces the sensitivity of the detection protocol for Salmonella,which could cause a negative deviation,resulting in the threshold exceeding of the Ac

关 键 词：沙门氏菌检验标准 微生物的确认方法 等效性 可接受限值 混合菌型污染食品 

分 类 号：R155[医药卫生—营养与食品卫生学]