SSR在猪个体识别及肉产品溯源中的应用

DNA溯源技术是根据动物个体之间遗传物质DNA序列的差异而进行个体识别并追溯到原产地的一种溯源技术。在试验群体中(11个品种,192个体)检测了24个SSR标记的遗传多样性,通过杂合度和多态信息含量计算筛选出11个SSR标记可用于猪肉产品的DNA溯源。在此基础上进一步在屠宰场采样进行了溯源模拟实验,结果表明筛选的11个SSR标记能区分100个个体,10份组织样品的SSR标记基因型是一一对应的,并且和43号个体基因型匹配。研究表明SSR标记可以用于猪个体识别和猪肉产品的溯源。     

利用10个微卫星标记对珠三角地区猪个体识别和肉类溯源

基于微卫星标记和毛细管电泳技术建立猪个体识别和肉类溯源系统,打击市场上来源不明的猪肉混合售卖的现象。采用10个微卫星标记进行PCR扩增和毛细管电泳分析,对珠三角地区采集的102个猪DNA样品进行个体识别和肉类溯源研究。结果表明：10个微卫星标记具有良好的猪个体识别和肉类溯源能力,多态性信息含量（PIC）在0.33和0.78之间,平均值为0.61;随机匹配概率（PM）和非父排除率（PE）的组合值分别为0.999999963514977和0.99592159278654;在实际应用中检出1例猪肉来源不明事件。该方法灵敏度高和准确性好,可应用于农场到市场各阶段的猪个体识别和肉类溯源,为监管部门提供技术支持。     

肉产品分子溯源标记的研究进展

食品溯源管理对食品生产、加工和消费过程起到重要的监管作用。肉产品作为饮食中主要的蛋白质来源,其安全问题受到高度重视。在众多的溯源技术方法中,本文介绍了DNA 溯源技术使用的分子标记的研究进展,对AFLP 标记、SSR 标记、SNP 标记3 类标记进行分析比较,并指出SNP 标记必将成为动物身份识别首选的分子标记。     

基于SNP标记的肉类溯源技术

本文阐述了建立肉类溯源系统对我国食品安全和国际贸易的重要性,并对溯源管理、食品溯源系统进行概述.重点讲述各种DNA标记技术的原理、利弊,以及在肉制品溯源标记应用的比较、SNP位点的检测方法等.最后指出了我国肉类溯源技术研究的方向和趋势.     

基于矿物元素指纹分析中国地理标志猪肉的产地溯源

本研究初步探讨了矿物元素指纹分析对我国地理标志猪肉产地溯源的可行性。采用电感耦合等离子体质谱对四川巴山的青峪黑猪肉,山东莱芜黑猪肉和北京黑六猪肉三种不同地域来源的地理标志猪肉中33种矿物元素含量进行测定。通过元素含量筛选,排除猪肉样品中含量低于或接近检出限的元素,筛选出13个元素进行研究。结合主成分分析、多重比较分析和判别分析对数据进行统计分析。其中Na、Fe、Co、Cu、Zn、Se、Rb、Sr共8种元素在地域之间差异显著(p<0.05)。通过对比偏最小二乘法判别分析(PLS-DA)、基于正交信号校正的偏最小二乘判别分析(OPLS-DA)、支持向量机、朴素贝叶斯、决策树和神经网络六种分类模型,得出OPLS-DA和决策树分类模型较适合基于矿物元素指纹分析的特色猪肉产地溯源。矿物元素指纹分析在我国地理标志猪肉产地溯源领域具有长远的应用前景。     

主成分分析技术对葡萄酒产地进行溯源

目的采用主成分分析技术对法国和山东产区的葡萄酒进行产地溯源。方法利用电感耦合等离子体质谱法(inductively coupled plasma mass spectroscopy,ICP-MS)、电感耦合等离子体发射光谱法(inductively coupled plasma emission spectrometry,ICP-OES)测定葡萄酒中30种无机元素及部分元素同位素含量,获取其中的元素成分信息,结合化学计量学中主成分分析技术(principal component analysis,PCA),分析不同地域样品的特征元素变量,研究筛选元素特征指纹。结果 PCA分析筛选出2个主成分因子,能对中国山东、法国波尔多产地的葡萄酒进行良好区分。利用PCA分析方法对中国山东、法国波尔多产区的葡萄酒样品的14种组分进行分析,筛选出~6Li、~7Li、~(10)B、~(11)B、Mg (280.270 nm)、P(213.618 nm)、Zn(213.857 nm)7种特征元素。结论该方法有望被应用于葡萄酒的产地溯源,可以为食品产地溯源技术的发展完善及相关部门的反欺诈监管提供技术积累。     

贝母属药用植物叶绿体基因组单核苷酸多态性位点生物信息学分析

目的分析贝母属药用植物叶绿体基因组单核苷酸多态性(single nucleotide polymorphism,SNP)位点的分布特点、鉴别能力和应用价值,为中药材贝母精准鉴定方法的建立提供技术支撑。方法应用多重序列比对、SNP筛选、酶切位点分析等生物信息学分析手段,对贝母属6种药用植物的15条叶绿体基因组序列进行分析。结果贝母属叶绿体基因组DNA同源性达98.38%,通过分析共发现SNP位点4058个,其中川贝母类鉴别候选位点71个,瓦布贝母鉴别候选位点个25个,太白贝母鉴别候选位点个120个,浙贝母鉴别候选位点个79个,湖北贝母鉴别候选位点个61个,平贝母鉴别候选位点794个。结论叶绿体基因组SNP分子标记因其密度高、鉴别力强和便于分析的特点,具有较大的应用价值。     

水产品产地溯源技术研究进展

水产品产地溯源技术是监管水产品质量安全的重要工具,不仅有利于保护消费者的合法权益,而且为保护地方特色产品提供了保障。近年来,质谱技术、光谱技术以及分子生物学技术等迅速发展,突显了其在水产品产地溯源中的重要作用,通过检测水产品的矿物元素组成与含量、同位素含量与比率、有机物组成、DNA图谱等信息,结合统计学分析手段,可建立水产品产地溯源指纹图谱。本文综述近年来常用的水产品产地溯源技术的应用研究进展,并对水产品产地溯源技术研究的发展趋势进行展望。     

代谢组学技术在食品产地溯源中的研究进展

食品产地溯源是建立产品质量安全追溯制度的重要环节之一,也是确保食品质量安全的必要手段。通过代谢组学技术分析产品的有机成分、同位素含量、有机化合物等特征成分,建立起能区分产品产地的溯源体系,增强消费者对食品安全的信心。综述了目前代谢组学技术在食品产地溯源中的应用研究情况,及不同技术各自的优缺点,并展望了今后代谢组学技术在食品产地溯源中的发展趋势。     

蜂蜜产地与蜜源矿物元素溯源方法的建立

基于矿物元素含量进行蜂蜜产地和蜜源溯源分析,建立产地和蜜源双目标溯源方法。采用全谱直读电感耦合等离子体原子发射光谱法(ICP-OES)测定蜜样矿物元素含量,在单目标溯源方法基础上,引进哑变量回归和校正模型,从而建立了蜂蜜产地、蜜源双目标溯源分析方法,并以四川蜂蜜为例,进行验证分析。结果显示,针对产地进行的逐步判别分析筛选出As、B、Ca等6个元素用于建立判别模型,回代验证的正确判别率达91.7%,交叉检验判别率为83.3%;针对蜜源的逐步判别分析筛选出B、Ca、Cu等5种元素用于建立判别模型,判别率为80%,交叉检验判别率为78.2%。基于矿物元素含量对蜂蜜进行产地与蜜源的溯源是可行的,所建立的方法具有一定借鉴意义;产地、蜜源两组判别方程适宜于研究区的溯源研究。     

Post-slaughter traceability

Traceability programs can cover the whole of life, or parts of it, for individual animals or groups/lots of animals. Of 13 country or community traceability programs for cattle/beef, 11 are mandatory (4 encompass, or are scheduled to encompass, birth to retail; 7 cover birth to slaughter) while 2 are voluntary and encompass birth to slaughter. Of 10 country or community traceability programs for swine/pork, 2 are mandatory (1 covers birth to retail; 1 covers birth to slaughter) while 8 are voluntary. Of 6 country or community traceability programs for sheep/sheep-meat, 3 are mandatory (1 encompasses birth to retail; 2 encompass birth to slaughter) while 3 are voluntary. Mandatory birth to retail programs that include "post-slaughter individual animal identification (IAID) traceability" have been implemented for cattle/beef, swine/pork and sheep/sheep-meat by the European Union and for cattle/beef by Japan. Many of the voluntary as well as mandatory, birth to slaughter traceability programs for all three species are presumed (though that is not specified) to include "post-slaughter group/lot identification (GLID) traceability" - e.g., those qualifying products for shipment to the European Union. "Post-slaughter IAID traceability" can be accomplished in very-small, small, medium, large and very-large packing plants using single-carcass processing units, tagging and separation/segregation, and/or deoxyribonucleic acid (DNA) fingerprinting technology but all of these approaches are time-consuming and costly; and, to-date, in most countries, there has been no reason compelling enough to cause industry to adopt such protocols or technology.     

Meat traceability using DNA markers: application to the beef industry

Consumer concerns about beef demands instruments to assure its traceability. A methodology using DNA markers is proposed for beef identification focussing on a Spanish beef certification, Ternera de Navarra (Beef of Navarra). To validate this methodology the number of markers used and the implications of population structure in individual identification were evaluated. In order to get practical implementation, the sampling levels required, depending on the number of markers and amount of possible fraud, is also discussed. Using at least eight very informative markers the origin of retailed meat is always found independent of genetic population structure. The total control of fraud would be very expensive using large-scale application of DNA analyses and a strategy based on anonymous sampling is proposed.     

Evaluation of microsatellites as a potential tool for product tracing of ground beef mixtures

Microsatellite genotyping was evaluated as a potential tool for DNA-based tracing of ground beef product. DNA from mixtures containing different numbers of individuals was analysed with a set of cattle microsatellite markers frequently used for parentage testing. As samples contained DNA from several animals, the microsatellite markers showed multiple peaks. The method could distinguish between mixtures containing equal amounts of meat from three different individuals, meat from three individuals mixed in different proportions, ground beef mixtures purchased in different cities, and different batches of ground beef patties. Limitations occurred when batches contained large numbers of individuals (>10) and different batches used meat from the same individuals. We conclude that DNA microsatellites may be useful for DNA traceability of ground beef mixtures prepared from less than 10 individuals, but where larger numbers of animals contribute to a mixture the method is not consistently accurate.     

基于RFID和条码技术的猪肉可追溯系统构建

基于RFID和条码技术,通过结合中间件技术、数据库技术、物流信息技术、数据传输技术等先进成熟的IT技术,建立了猪肉可追溯系统,实现肉类食品质量安全全程跟踪与追溯。该系统可以有选择的进行批次与个体追溯。     

Genetic control of conventional labeling through the bovine meat production chain by single nucleotide polymorphisms using real-time PCR

Since January 2002, the European Union has adopted precise guidelines aimed at protecting the safety of meat and controlling the production chain. To this purpose, the conventional traceability of livestock and meat represents the main tool, but verification of traceability requires genetic support. At present, single nucleotide polymorphisms (SNPs) represent the most innovative molecular markers in genotyping studies. The aim of this study was to verify correct labeling in a bovine meat production chain by a real-time PCR protocol based on SNP analysis. Reference hair samples from 5,000 animals were randomly collected from 22 farms. Twelve hundred meat samples were collected at different steps of the bovine meat production chain. In particular, 1,000 meat samples were collected at the slaughterhouse and 200 samples from the same animals directly at the butcher's shop. The protocol was optimized and validated by testing a set of 16 SNP markers on 95 DNA samples from bovine sires of different breeds. Thereafter, the genotyping of 2,200 samples was conducted with a set of 12 selected SNPs to verify traceability of the meat production chain at three different stages: farm, slaughterhouse, and butcher's shop. Irregularities in conventional traceability were evidenced directly in 1.87% of the samples at the slaughterhouse. This percentage increased to 3.25% when sampling was conducted at the butcher's shop. This study demonstrates that despite the precautions adopted over the meat production chain, some critical points still exist that cause the loss of a correct association between registration numbers and samples.     

基于贝叶斯网络的猪肉合格率的模型构建

我国是猪肉消费大国,其卫生情况对人民关系重大。近年来肉制品的微生物污染现象不容乐观,所以建立一个行之有效的猪肉追溯系统有利于更好地控制猪肉质量。本文根据现有的猪肉卫生标准,选出猪肉的四种优势致腐菌:大肠杆菌、单增李斯特菌、沙门氏菌以及金黄色葡萄球菌,以贝叶斯网络为理论依据,利用Hugin Lite软件拟合出合格率模型。结果表明,基于贝叶斯网络可通过改变菌体数据快速求得猪肉的合格率,大大缩减检验时间,为猪肉卫生合格率的质检工作提供理论依据。     

A false single nucleotide polymorphism generated by gene duplication compromises meat traceability

Controlling meat traceability using SNPs is an effective method of ensuring food safety. We have analyzed several SNPs to create a panel for bovine genetic identification and traceability studies. One of these was the transversion g.329C>T (Genbank accession no. AJ496781) on the cytochrome P450 17A1 gene, which has been included in previously published panels. Using minisequencing reactions, we have tested 701 samples belonging to eight Spanish cattle breeds. Surprisingly, an excess of heterozygotes was detected, implying an extreme departure from Hardy-Weinberg equilibrium (P<0.001). By alignment analysis and sequencing, we detected that the g.329C>T SNP is a false positive polymorphism, which allows us to explain the inflated heterozygotic value. We recommend that this ambiguous SNP, as well as other polymorphisms located in this region, should not be used in identification, traceability or disease association studies. Annotation of these false SNPs should improve association studies and avoid misinterpretations.