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低温乳制品中沙门氏菌实时荧光PCR检测方法体系的建立

来源:用户上传      作者:陶文靖 赵婷婷 王琦 董彬 王玉花 王维嘉 刘赫东 曲连海

  摘 要:低温乳制品保质期短,对运输、储存条件要求较高,如果储存温度未达到要求,就会增加微生物增殖的风险。沙门氏菌作为乳制品必检项目需要重点关注,但现有国家标准检测方法操作较烦琐,耗时长,无法满足快速检测需求;国内现行有效的沙门氏菌实时荧光聚合酶链式反应(Polymerase Chain Reaction,PCR)行业标准方法,操作环节复杂,对人员操作要求较高。本研究通过优化增菌培养基以及实时荧光PCR方法,建立了一套更严格、简单、适合企业操作的沙门氏菌实时荧光PCR方法体系,并经过科学的验证评价,以满足低温乳制品生产加工过程及终产品快速放行、及时纠偏的需求,提高致病菌检测技术能力,降低食品安全风险。
  关键词:低温乳制品;沙门氏菌;实时荧光PCR;方法评价
  Establishment of a Real-time Fluorescent PCR Method for Detection of Salmonella in Low Temperature Dairy Products
  TAO Wenjing1, ZHAO Tingting2, WANG Qi3, DONG Bin4, WANG Yuhua3, WANG Weijia5, LIU Hedong1, QU Lianhai1*
  (1.Beijing Meizheng Bio-Tech Co., Ltd., Beijing 102200, China;
  2.Bright Dairy Co., Ltd., East China Center Factory, Shanghai 201111, China;
  3.SGS-CSTC Standards Technical Services (Qingdao) Co., Ltd., Qingdao 266071, China;
  4.Tianjin Bright & Mengde Dairy Co., Ltd., Tianjing 300499, China;
  5.Wuhan Branch, SGS-CSTC Standards Technical Services Co., Ltd., Wuhan 430056, China)
  Abstract: Low temperature dairy products have short shelf life and high requirements for transportation and storage conditions. If the storage temperature does not meet the requirements, it will increase the risk of microbial proliferation. Salmonella, as a mandatory item of dairy products, needs special attention, but the existing national standard detection methods are cumbersome and time-consuming, and can not meet the needs of rapid detection; The current effective Salmonella real-time fluorescent polymerase chain reaction (PCR) industry standard method in China has complex operation links and high requirements for personnel operation. By optimizing the enrichment medium and the real-time fluorescent PCR method, this study has established a more rigorous, simple and suitable real-time fluorescent PCR method system for Salmonella for enterprise operation, which has been scientifically verified and evaluated to meet the needs of the production and processing process of low-temperature dairy products and the rapid release and timely correction of end products, improve the technical ability of pathogenic bacteria detection, and reduce food safety risks.
  Keywords: low-temperature dairy products; Salmonella; real-time fluorescence PCR; method evaluation
  近年恚我国居民对营养健康的关注度越来越高,对牛奶可以提升免疫力的认可度较高[1]。液态乳制品中低温鲜奶以“更新鲜、更营养”的特点受到消费者的青睐。《2020中国奶商指数报告》显示,96.0%的中国消费者认为低温鲜奶富含乳铁蛋白和免疫球蛋白等活性因子,能有效提升机体免疫力[2]。
  近几年,国产优质低温巴氏杀菌乳的灭菌温度已由105 ℃降低到75 ℃,乳中含有的乳铁蛋白、免疫球蛋白、过氧化物酶等对人体有益的活性成分得到最大限度的保留,可以达到进口乳制品的8倍[3-4]。这种杀菌工艺杀灭了大部分微生物,允许残留部分微生物,将微生物含量降低到不会威胁人体健康的程度[5]。低温乳制品保质期短,根据包装形式的不同,在2~6 ℃条件下,巴氏杀菌乳保质期一般为2~7 d,低温酸奶保质期一般为14~21 d[6]。巴氏杀菌乳对运输、储存条件要求较高,一旦储存温度达不到要求,可能会存在微生物增殖的风险[7]。食源性致病菌是引起食品安全问题的主要因素之一,而微生物污染是导致牛奶质量问题的重要因素,因此牛奶中的致病菌成为消费者关注的焦点[8-10]。根据我国国标要求,需重点关注的致病性微生物有金黄色葡萄球菌、沙门氏菌等。

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