转基因水稻定性PCR检测体系的建立

采用PCR技术检测CaMV35S启动子、NOS终止子、Bt基因,判断水稻样品中是否含转基因成分,建立了稳定快速的转基因水稻定性PCR检测体系.PCR检测的灵敏度达到0.1%,稳定性良好.结果表明:PCR技术检测外源基因是灵敏和准确的,可以广泛地应用到转基因作物及其加工产品的转基因成分检测中.此外,进一步讨论了转基因成分检测中存在的问题、转基因产品标识的必要性和目前转基因检测技术在检测中的问题.     

利用实时荧光PCR方法检测转Bt基因大米

应用实时荧光PCR技术对转基因大米进行了定性和定量检测研究.本研究以转基因B163大米为材料,采用TaqMan探针技术,对大米中的内源基因蔗糖磷酸合酶SPS和转基因水稻中普遍存在的外源基因CaMV35S启动子、NOS终止子以及苏云金芽孢杆菌(Bacillusthndngiemis,简写为Bt)杀虫晶体蛋白基因Cry1Ac进行了实时荧光PCR研究,并对外源基因Cry1Ac进行了定量检测和敏感性分析.该实时荧光PCR方法检测结果和常规PCR结果一致,同时不用进行凝胶电泳,更为快速、简便,降低了污染机会,可用于转Bt基因大米的定性和定量检测.     

转基因稻米加工食品的PCR检测技术研究

本文对转基因稻米及其加工食品进行了针对标记基因bar、CaMV35S启动子和Nos终止子DNA序列的PCR检测.结果表明,改良SDS法适于提取转基因稻米以及经过蒸煮、微波膨化加工后稻米食品中的DNA并能够满足PCR检测的需要.食品加工使稻米基因组DNA降解为较小的片段,但不影响PCR检测效果,PCR技术能够检测稻米加工食品中的转基因成分.     

双抗夹心ELISA检测转Bar基因抗除草剂大豆

为快捷有效地检测转Bar基因抗除草剂大豆,利用已制备的抗除草剂Bar基因编码蛋白,膦丝菌素乙酰转移酶（phosphinothricin acetyltransferase,PAT）单克隆抗体和多克隆抗体,建立PAT蛋白双抗夹心酶联免疫吸附检测方法,对转Bar基因抗除草剂大豆不同组织材料进行定量检测。结果显示,最佳检测条件为捕获抗体质量浓度0.125 μg/mL,包被酶标板,37 ℃孵育1 h后4 ℃静置过夜,检测样品37 ℃孵育1.5 h,检测抗体质量浓度6.25 μg/mL,37 ℃孵育1.5 h;PAT蛋白的最低检测限为0.04 ng/mL,大豆蛋白体系中为8 ng/mL;重复性变异系数小于3%。利用上述检测条件,对实验建立的转Bar基因抗除草剂大豆进行PAT蛋白定量检测,成功地在根、茎、花、叶、种子不同部位检测到该蛋白的表达。     

多重PCR检测4种常见食源性致病菌

目的 建立一种多重聚合酶链式反应法(multiplex polymerase chain reaction, MPCR)快速检测肉制品中金黄色葡萄球菌、沙门氏菌、志贺氏菌和单增李斯特氏菌的分析方法。方法 选取金黄色葡萄球菌nuc基因、沙门氏菌SipB基因、志贺氏菌ipaH基因、单增李斯特菌inlA基因作为目标基因, 设计4对PCR引物, 建立并优化多重PCR反应体系, 评价该体系的特异性和灵敏度, 并对人工污染的熟肉样品进行检测。结果 构建的多重PCR方法特异性强、灵敏度高, 人工污染熟肉匀浆中4种致病菌的检出限为103 CFU/mL。结论 构建的多重PCR检测方法能够快速、准确、高效地检测肉制品中金黄色葡萄球菌、沙门氏菌、志贺氏菌和单增李斯特氏菌, 为食源性疾病菌的快速检测提供参考依据。     

鉴定金黄色葡萄球菌的多重PCR方法

建立了一种快速鉴定和鉴别金黄色葡萄球菌的多重PCR方法。利用金黄色葡萄球菌的特异基因、耐甲氧西林基因和4种肠毒素基因nuc、mecA和sea、sec、sed、see建立6重PCR反应体系,并对PCR体系,引物浓度进行优化。6对PCR引物均能特异地扩出相应的目的基因。对47株金黄色葡萄球菌的检测结果与应用单重PCR鉴定结果完全一致。实验所建立的多重PCR方法能在一个反应体系中鉴定和鉴别金黄色葡萄球菌,为食品中金黄色葡萄球菌的快速检测提供了一种有效的检测手段。     

大豆加工食品中转基因成分多重PCR检测体系的建立

以大豆内源基因 (Lectin)、筛选基因 3 5S启动子 (Cauliflowermosaicvirus 3 5S ,CaMV3 5S)、Nos终止子 (Nopalinesynthase,Nos)和外源基因 (5 enolpyruvylshikimate 3 phosphatesynthase ,Ep sps)为检测对象 ,通过对PCR扩增体系中各引物终浓度及PCR扩增过程中退火温度的探讨 ,研究了不同引物终浓度配比及退火温度对转基因大豆多重PCR检测的影响 ,建立了大豆加工食品中转基因成分多重PCR检测体系。结果表明 ,当各组引物的终浓度分别为 1 0、2 0、2 0、3 0 μmol/L即引物终浓度配比为 1∶2∶2∶3 ,退火温度为 5 5 4℃时 ,所建立的多重PCR检测方法能够有效地检测出大豆中的转基因成分 ,具有特异性好 ,简便 ,快速 ,准确等优点。     

利用PCR方法检测转BT基因水稻

应用PCR技术进行转基因水稻检测研究。本文以转基因水稻为材料，以聚合酶链式反应（PCR）方法为基础，选择适用于转基因食品安全性检验的核酸检测技术，针对转基因水稻中普遍存在的花椰菜花叶病毒（CaMV35S）启动子、胭脂碱合成酶（NOS）终止子、和转入苏云金杆菌（Bacillus thuringiensis，简写为Bt）基因水稻的CrylAc片段进行PCR检测，建立适合转BT基因水稻的检测方法。该方法简便快速、检测结果与标准及其他文献资料相符。     

基于16S rRNA基因序列鉴定秘鲁茎柔鱼和澳洲双柔鱼

通过对秘鲁茎柔鱼及澳洲双柔鱼的线粒体16S rRNA基因片段进行聚合酶链式反应（polymerase chain reaction,PCR）扩增和序列比对,确定2 种柔鱼品种的特异性位点,并由此设计品种特异性引物。利用多重PCR体 系对秘鲁茎柔鱼和澳洲双柔鱼进行快速、准确地鉴定和区分,并对该体系的灵敏度进行检验。结果表明：在多重 PCR体系下,秘鲁茎柔鱼和澳洲双柔鱼分别扩增出长度400、229 bp的品种特异性条带,2 种柔鱼的混合样品则同时 扩增出长度400、229 bp的条带;且该多重PCR体系可以检测澳洲双柔鱼中1%秘鲁茎柔鱼的掺入,检测限为0.1 ng。     

3种致病菌多重PCR检测体系的建立及应用

金黄葡萄球菌(Staphylococcus aureus,SA)、单核细胞增生性李斯特菌(Listeria monocytogen,LM)和蜡样芽孢杆菌(Bacillus cereus,BC)是食品中重要的致病菌,建立其多重PCR的快速检测体系,对开发食源性致病微生物快速检测试剂盒具有重要意义。根据SA的nuc基因、LM的hly基因和BC的hemolysin基因,设计合成3对特异性引物,然后进行单基因PCR反应特异性验证。在此基础上建立了3种致病菌的多重PCR检测体系,并应用于食品检测中,同时以国标法进行对比验证。结果表明,建立的多重PCR检测方法简单、快速、灵敏度高,检测灵敏度可达到1 cfu/mL,整个检测时间在16 h以内,具有较大的应用价值,可广泛应用于食品卫生检测以及临床检验等领域。     

Reliable identification of grapevine rootstock varieties using RAPD PCR on woody samples

Since grapevine ( Vitis spp .) rootstock material is being traded increasingly as disbudded woody material a lack of distinctive morphological features on such material necessitates an alternative and reliable means of identification. Methods described here were developed for rapid and efficient extraction of DNA from woody samples rich in phenolic compounds and polysaccharides, and for subsequent identification of varieties by RAPD PCR. Using these methods, and with the application of only one selected RAPD primer, we were able to differentiate sixteen rootstock varieties, including the seven varieties most commonly used in Germany. Problems commonly encountered with reproducibility of RAPD patterns were avoided by choosing primers with a dinucleotide sequence and a high G/C content that allowed a rather high annealing temperature of 45°C. Methods described here should also be useful for other horticultural crops, especially those with woody tissues rich in phenolic compounds and polysaccharides.     

An internet compendium of analytical methods and spectroscopic information for monomers and additives used in food packaging plastics

An internet website (http://cpf.jrc.it/smt/) has been produced as a means of dissemination of methods of analysis and supporting spectroscopic information on monomers and additives used for food contact materials (principally packaging). The site which is aimed primarily at assisting food control laboratories in the European Union contains analytical information on monomers, starting substances and additives used in the manufacture of plastics materials. A searchable index is provided giving PM and CAS numbers for each of 255 substances. For each substance a data sheet gives regulatory information, chemical structures, physico-chemical information and background information on the use of the substance in particular plastics, and the food packaging applications. For monomers and starting substances (155 compounds) the infra-red and mass spectra are provided, and for additives (100 compounds); additionally proton NMR are available for about 50% of the entries. Where analytical methods have been developed for determining these substances as residual amounts in plastics or as trace amounts in food simulants these methods are also on the website. All information is provided in portable document file (PDF) format which means that high quality copies can be readily printed, using freely available Adobe Acrobat Reader software. The website will in future be maintained and up-dated by the European Commission's Joint Research Centre (JRC) as new substances are authorized for use by the European Commission (DG-ENTR formerly DGIII). Where analytical laboratories (food control or other) require reference substances these can be obtained free-ofcharge from a reference collection housed at the JRC and maintained in conjunction with this website compendium.     

Aroma characterization of various apricot varieties using headspace–solid phase microextraction combined with gas chromatography–mass spectrometry and gas chromatography–olfactometry

The characterization of the aromatic profile of several apricot cultivars with molecular tracers in order to obtain objective data concerning the aromatic quality of this fruit was undertaken using headspace–solid phase microextraction (HS–SPME). Six apricot cultivars were selected according to their organoleptic characteristics: Iranien, Orangered, Goldrich, Hargrand, Rouge du Roussillon and A4025. The aromatic intensity of these varieties measured by HS–SPME–Olfactometry were defined and classified according to the presence and the intensity of grassy, fruity and apricot like notes. In the six varieties, 23 common volatile compounds were identified by HS–SPME–GC–MS. Finally, 10 compounds, ethyl acetate, hexyl acetate, limonene, β-cyclocitral, γ-decalactone, 6-methyl-5-hepten-2-one, linalool, β-ionone, menthone and (E)-hexen-2-al were recognized by HS–SPME–GC–O as responsible of the aromatic notes involved in apricot aroma and considered as molecular tracers of apricot aromatic quality which could be utilized to discriminate apricot varieties.     

Tests of potential functional barriers for laminated multilayer food packages. Part I: Low molecular weight permeants

The advent of the functional barrier concept in food packaging has brought with it a requirement for fast tests of permeation through potential barrier materials. In such tests it would be convenient for both foodstuffs and materials below the functional barrier (sub-barrier materials) to be represented by standard simulants. By means of inverse gas chromatography, liquid paraffin spiked with appropriate permeants was considered as a potential simulant of sub-barrier materials based on polypropylene (PP) or similar polyolefins. Experiments were performed to characterize the kinetics of the permeation of low molecular weight model permeants (octene, toluene and isopropanol) from liquid paraffin, through a surrogate potential functional barrier (25 μm-thick oriented PP) into the food simulants olive oil and 3% (w/v) acetic acid. These permeation results were interpreted in terms of three permeation kinetic models regarding the solubility of a particular model permeant in the post-barrier medium (i.e. the food simulant). The results obtained justify the development and evaluation of liquid sub-barrier simulants that would allow flexible yet rigorous testing of new laminated multilayer packaging materials.     

Chlorohydrins of bisphenol A diglycidyl ether (BADGE) and of bisphenol F diglycidyl ether (BFDGE) in canned foods and ready-to-drink coffees from the Japanese market

BADGE.2HCl and BFDGE.2HCl were determined in 28 samples of ready-to-drink canned coffee and 18 samples of canned vegetables (10 corn, 5 tomatoes and 3 others), all from the Japanese market. HPLC was used as the principal analytical method and GCMS for confirmation of relevant LC fractions. BADGE.2HCl was found to be present in one canned coffee and five samples of corn, BFDGE.2HCl in four samples of canned tomatoes and in one canned corn. No sample was found which exceeded the 1mg/kg limit of the EU for the BADGE chlorohydrins. However the highest concentration was found for the sum of BFDGE.2HCl and BFDGE.HCl.H₂O at a level of 1.5mg/kg. A Beilstein test confirmed that all cans containing foods contaminated with BADGE.2HCl or BFDGE.2HCl had at lest one part coated with a PVC organosol.     

European priorities for research to support legislation in the area of food contact materials and articles

A strong science base is required to underpin the planning and decision-making process involved in determining future European community legislation on materials and articles in contact with food. Significant progress has been made in the past 5 years in European funded work in this area, with many developments contributing to a much better understanding of the migration process, and better and simpler approaches to food control. In this paper this progress is reviewed against previously identified work-areas (identified in 1994) and conclusions are reached about future requirements for R&D to support legislation on food contact materials and articles over the next 5 or so years.     

Analysis of benzo[a]pyrene in spiked fatty foods by second derivative synchronous spectrofluorimetry after microwave-assisted treatment of samples

A simple, rapid and inexpensive method has been developed for the determination of benzo[a     

The development of reference materials for paralytic shellfish poisoning toxins in lyophilized mussel. II: Certification study

This paper describes the second part of a project undertaken to develop certified mussel reference materials for paralytic shellfish poisoning toxins. In the first part two interlaboratory studies were undertaken to investigate the performance of the analytical methodology for several PSP toxins, in particular saxitoxin and decarbamoyl-saxitoxin in lyophilized mussels, and to set criteria for the acceptance of results to be applied during the certification exercise. Fifteen laboratories participated in this certification study and were asked to measure saxitoxin and decarbamoyl-saxitoxin in rehydrated lyophilized mussel material and in a saxitoxin-enriched mussel material. The participants were allowed to use a method of their choice but with an extraction procedure to be strictly followed. The study included extra experiments to verify the detection limits for both saxitoxin and decarbamoyl-saxitoxin. Most participants (13 of 15) were able to meet all the criteria set for the certification study. Results for saxitoxin.2HCl yielded a certified mass fraction of <0.07 mg/kg in the rehydrated lyophilized mussels. Results obtained for decarbamoyl-saxitoxin.2HCl yielded a certified mass fraction of 1.59+/-0.20 mg/kg. The results for saxitoxin.2HCl in enriched blank mussel yielded a certified mass fraction of 0.48 +/- 0.06 mg/kg. These certified reference materials for paralytic shellfish poisoning toxins in lyophilized mussel material are the first available for laboratories to test their method for accuracy and performance.     

Announcing a new international peer-reviewed journal:Food Science and Human Wellness now accepting manuscript submission in English

<正>We are pleased to announce the launch of a new international peer-reviewed journal-Food Science and Human Wellness,ISSN 2213-4530,which is an open access journal,produced and hosted by Elsevier B.V.on behalf of Beijing Academy of Food Sciences.Food Science and Human Wellness is an international peer-reviewed English journal that provides a forum for the dissemination of the