Multiplex PCR system to track authorized and unauthorized genetically modified soybean events in food and feed

A diverse range of genetic elements has been used to develop genetically modified organisms (GMOs) over the last 18 years. Screening methods that target few elements, such as the Cauliflower Mosaic Virus 35S promoter (P-35S) and Agrobacterium tumefaciens nopaline terminator (T-nos), are not sufficient to screen GMOs. In the present study, a multiplex PCR system for all globally commercialized GM soybean events was developed to easily trace the events. For this purpose, screening elements of 24 GM soybean events were investigated and 9 screening targets were selected and divided into three individual triplex PCR systems: P-35S, ribulose-1,5-bisphosphate carboxylase small subunit promoter of Arabidopsis thaliana, T-nos, T-35S, pea E9 terminator, open reading frame 23 terminator of A. tumefaciens, proteinase inhibitor II terminator of potato, acetohydroxy acid synthase large subunit terminator of A. thaliana, and the revealed 3′ flanking sequences of DP-305423-1. The specificity of the assays was confirmed using thirteen GM soybean events as the respective positive/negative controls. The limit of detection of each multiplex set, as determined using certified reference materials of specific GM events, ranged from 0.03 to 0.5%, depending upon target. Furthermore, 26 food samples that contained soybean ingredients, which were purchased from the USA, China, Japan, and Korea, were analyzed, 17 of which contained one or more GM soybean events. These results suggest that the developed screening method can be used to efficiently track and identify 24 GM soybean events in food and feed.     

高强度低合金铌钒钢板的失效分析

针对高强度低合金铌钒钢板在拉伸试验后出现分层开裂的现象,采用SEM、EDAX进行微观分析,发现试样内部有大量夹杂物存在,这些夹杂物主要含有钙、氧、硫、锰等元素。带有缺陷的钢板若应用到实际工程中,会影响铁塔的寿命。     

基于神经网络的氟石粉液化模型分析

在对氟石粉进行实验研究的基础上,应用RBF神经网络,建立了预测氟石粉液化FMP的模型。在该模型中,输入为影响氟石粉液化FMP的各种因素,包括粒径、均匀度、化学性质、真密度及堆积密度等,输出为FMP,预测结果表明,所建RBF神经网络模型预测精度较高,可应用于对氟石粉液化FMP的预测。同时采用逐步反向删除的方法分析输入属性对输出结果的影响,找出了影响氟石粉液化FMP的主要影响因素。最后对每个主要属性进行了灵敏度分析。     

Efficiency of acetic acid and formic acid as a catalyst in catalytical and mechanocatalytical pretreatment of barley straw

In this study, the potential of organic acids (formic acid, acetic acid) in a catalytical and mechanocatalytic conversion of lignocellulosic barley straw to valuable sugars is explored using sulfuric acid as a reference. Acid-catalyzed hydrolysis has been carried out with acid-impregnated samples as well as unmodified barley straw. In the mechanocatalytical approach, pretreatment consists of impregnation with the acid catalyst and mechanical treatment by ball milling following chemical hydrolysis. Straw samples and residues were analyzed by Fourier transform infrared spectrometry (FT-IR) whereas hydrolysate analysis was based on total reducing sugar (TRS) determination following the DNS method and capillary electrophoresis (CE) analysis. The results indicated that acetic acid and formic acid are rather mild acids yielding low TRS levels compared to the reference acid. Mechanocatalytical pretreatment slightly increased TRS yields, but not significantly. Strikingly, sulfuric acid showed an efficient conversion efficiency yielding almost 45% of TRS. Furthermore, this study provided evidence for the acetylation of straw components when acetic acid was used as catalyst. Alkali hydrolysis induced the de-esterification, but revealed no significant increase of TRS yields.     

Low temperature cleanup combined with magnetic nanoparticle extraction to determine pyrethroids residue in vegetables oils

To quantify trace pesticide residue in vegetable oil rapidly, low temperature cleanup combined with magnetic nanoparticle based solid phase extraction was developed to determine eight pyrethroids in vegetable oils, including tetramethrin, fenpropathrin, cypermethrin, decamethrin, fenvalerate, acrinathrin, permethrin and bifenthrin. Polystyrene coated magnetic nanoparticles were synthesised by a modified chemical coprecipitation combined with emulsion polymerisation method. The nanoparticles were afterwards characterised by Fourier transform-infrared spectroscopy, X-ray diffraction, transmission electron microscopy as well as vibrating sample magnetometer, and successfully employed as adsorbents for the magnetic solid phase extraction of pyrethroids which were cleaned up using low temperature approach in advance. Critical impact factors on the efficiency of the extraction method such as the mass of adsorbents used, volume and type of eluent solvent, extraction time as well as elution time were optimised subsequently. Regression analysis of the calibration curves of the eight pyrethroids yielded satisfactory correlation coefficients within the range of 0.980–0.998. Limit of detection and limit of quantification were calculated to be between 0.0290-0.0658 and 0.0890–0.1994 ng g⁻¹, respectively. Intra-day and inter-day reproducibility at different concentration levels also produced satisfactory recovery rates of 83.18–112.79% with relative standard deviations not exceeding 10.84% and 12.01%, respectively, suggesting desirable stability of the proposed method.     

Influence of MWCNTs modified by silane coupling agent KH570 on the properties and structure of MWCNTs/PLA composite film

Composite films based on poly(lactic acid)(PLA) filled with modified multiwalled carbon nanotubes (MWCNTs) were prepared by casting/solvent evaporation technique and investigated for their properties and structures. MWCNTs were modified by coupling agent KH570 which can improve the dispersion of MWCNTs and then enhance the comprehensive properties of PLA. The influences of modified conditions, including the amounts of KH570, the reaction temperature and time, were analyzed. And also the structures and properties of composite films were characterized by X-ray diffraction (XRD), scanning electron microscopy (SEM), fourier transform infrared (FTIR) and thermogravimetric analysis (TG). The two components of the composite films, WMCNTs and PLA, had excellent compatibility. The addition of MWCNTs modified by coupling agents KH570 was beneficial for the crystallization of PLA, and the toughness of composite films was also improved. When the reaction temperature was 70 °C, reaction time was 6 h, and the amount of KH570 was 3 % respectively, the obtained composite films had the perfect comprehensive properties.     

带有磺酸基团和羰基酸团的新固体酸的合成及其在缩醛化反应中的催化性能简

The novel solid acid with both sulfonic and carbonyl acid groups has been synthesized from 3-（（2-sulfoethoxy） carbonyl）acrylic acid and tetraethyl orthosilicate （TEOS）. The catalytic activities were investigated through the acetalization. The results showed that the novel solid acid was very efficient for the reactions with the high yields. The high acidity, high stability and reusability were the key feature of the novel solid acid. Moreover, the sulfonic and carbonyl acid groups could cooperate during the catalytic process, which improved its catalytic activities. The catalyst shows recyclability, and hold great potential for replacement of homogeneous catalysts.     

单增李斯特菌毒力因子多重荧光PCR法 建立与应用

目的建立同时检测单增李斯特菌(Listeria monocytogenes)及其3种毒力因子的多重荧光PCR快速检测方法,并应用于日常食品的检测。方法根据单增李斯特菌溶血素基因hly A、内化素基因inl A和表面蛋白act A基因的保守序列,分别设计合成特异性引物和探针,优化多重荧光PCR反应体系。对该方法的特异性、敏感性和重复性进行评估。结果该法特异性强、敏感性高,对单增李斯特菌纯培养物的最低检出限410cfu/m L;重复性好,变异系数均小于2%。对84份食品检测结果与传统国标法相符,共检出单增李斯特菌4份,检出率为4.76%。多重荧光PCR检测方法耗时1 h,比传统方法节约2~5 d。4株单增李斯特菌分离株中2株同时含有inl A、act A、hly A 3种毒力基因,另2株为毒力基因act A缺失株,提示目前流行株并非同一来源。结论本研究建立的多重实时荧光PCR方法能同时对单增李斯特菌及其3种毒力因子进行快速检测,且灵敏度高、特异性好,为食源性疾病的病原学检测提供了快速可靠的方法。     

食品过敏原羽扇豆成分的实时荧光PCR检测方法

羽扇豆是欧盟法规标签要求强制性标识的食品过敏原。采用现有参考文献TaqMan实时荧光PCR方法,研究建立食品过敏原羽扇豆成分的检测方法。研究表明建立的研究方法具有特异性,灵敏度高,检测限为1 mg/kg。此方法适用于食品中过敏原羽扇豆成分检测。     

进口油菜籽中不同转基因品系的检测与分析

对我国27批进口油菜籽进行了转基因成分检测和转基因油菜品系组成情况分析。利用已知转基因油菜RT73、T45、Oxy235、Topas19/2、MS1、RF1、RF2、MS8和RF3的特异性检测体系,对初检为阳性的转基因油菜籽样品进行扩增,扩增基因包括bar、PAT、GOX和NPTII。结果共检出25批转基因油菜籽,检出率为92.6%。所有批次均检出品系RT73和MS8×RF3,表明二者为目前我国进口的主要转基因油菜籽。约40%的批次检出MS1×RF1和MS1×RF2,有少量批次检出T45、Oxy235和Topas19/2,每批次均为多品系混合样品。研究表明,列入农业转基因生物安全生产证书（我国农业部颁发）名单的7个转基因油菜品系均有进口,其中以RT73和MS8×RF3为主。