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细菌数值鉴定法及其在食源性李斯特菌中的应用
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摘要
细菌数值鉴定法是细菌鉴定学和数学统计学的结合,它将统计学中的概率值应用到细菌的鉴定试验中,通过比较待测菌株与细菌数据库中已知菌株的生化反应的符合概率,从而得到鉴定结果。本文对细菌数值鉴定法的鉴定原理、生化反应的选择和编码过程进行了介绍,并通过此方法筛选了11种不同于目前市面上同等产品的生化试验组合。对本实验室前期分离到近2000株李斯特菌进行相关实验,获得各生化反应阳性率,建立了这11种生化反应阳性率数据库。运用C++语言等计算机技术编写相关软件,初步枸建了李斯特菌生化鉴定系统(Numide-Listeria v1.0),继而对从全国食品中分离的880株疑似李斯特菌分别进行了MID-67、Numide-Listeria v1.0和分子鉴定比较,分子鉴定结果显示99.8%的菌株含有种特异性基因,而这其中99.3%的单增李斯特菌、96.3%的英诺克李斯特菌、100%的格式李斯特菌、100%的西式李斯特菌、100%的绵羊李斯特菌和80%威尔士李斯特菌显示两种鉴定系统结果一致,而本论文研制的Numide-Listeria v1.0在鉴定单增李斯特菌和英诺克李斯特菌方面表现出优于MID-67的能力。Numide-Listeria v1.0的初步构建,为下一步探索更为特异的生化反应,建立新型李斯特菌快速检测体系提供了理论依据和技术支持。
Numerical method of bacterial identification is one of probabilistic identification of bacteria which generated by the combination of the bacterial identification studies and mathematical statistics.The identification result found on unknown strains can be matched with those stored for previously strains which get the probabilityof giving positive result in each test.Thisresearch will introduce the theory of identification,choice of tests and the numerical code identification schemes for the numerical method of bacterial identification.Based on the different biochemistry results among Listeria spp.,eleven tests were selected which were differ with market products.The percentages of positives reactions of these strains were obtained from approximately 2000 Listeria strains which isolated from previous study in ourlaboratory.Combining computer technology,we have programmed Numide-Listeria vl.0 by C + + language.Numide-Zisterta vl.0 could distinguish six species of listeria in food effectively.Besides,a total of 880 Listeria isolates were identified by MID-67,Numide-Zisteria v1.0 and PCR method.Of which,99.3%of L.monocytogenes,96.3%of L.innocua,100%of L.grayi,100%of L.seeligeri,100%of L.ivanovii and 80%of Listeria welshimeriv/ere consistent with three methods.It also showed higher identified ability for L.moocytogenes and L innocua by our numerical identification system.It is benefit for the improvement for biochemical identification system of Listeriaspp.
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