共查询到17条相似文献，搜索用时 156 毫秒
1.
《中国油料作物学报》2016,(5)
为提高植物油中真菌毒素的检测效率,研制免疫亲和柱,建立检测方法。以纯化的抗黄曲霉毒素(AFT,包括AFB_1、AFB_2、AFG_1、AFG_2)、抗玉米赤霉烯酮毒素(ZEA)、抗T-2毒素、抗伏马B1毒素(FB_1)的四种单克隆抗体为特异性抓捕元件,通过研究比较3种载体材料和2种抗体偶联方法,成功制备出以CNBr-activated Crystarose4B作为固相载体材料的真菌毒素多组分免疫亲和柱。同时研究比较了提取溶剂、提取方式和上样缓冲液等条件,并结合高效液相色谱串联质谱技术(HPLC-MS/MS),建立了植物油中7种真菌毒素的同步免疫亲和净化——HPLC-MS/MS检测方法。结果表明,该方法对7种真菌毒素的检测线性范围宽,线性回归方程相关系数(R2)均不低于0.992;对AFB_1、AFB_2、AFG_1、AFG_2四种真菌毒素的检出限为0.02~0.08μg/kg,对ZEA、T-2、FB_1三种真菌毒素的检出限为0.10~0.50μg/kg;添加回收实验中,低、中、高三个不同加标水平下的平均回收率为80.22%~106.10%,相对标准偏差为1.05%~6.56%。该方法操作简便、净化彻底、回收率高、有机溶剂消耗量少,为食用植物油中真菌毒素多组分同步确证性检测提供了一种高效、准确、灵敏的技术。 相似文献
2.
纳米抗体是由骆驼科动物产生的一类天然缺失轻链的重链抗体可变区.相比较于传统单克隆抗体和多克隆抗体,其具有体积小、热稳定性强、耐有机溶剂、折叠性可逆、易于表达、高亲和力、可特异性识别独特表位等天然优势.因此,纳米抗体在生物和农业等研究领域备受关注.本文综述了纳米抗体的结构、生化特性以及在农产品真菌毒素检测中的研究进展,并... 相似文献
3.
完善的实验室能力验证体系是确保真菌毒素检测机构出具准确检测结果的重要前提。本文重点阐述了国内外真菌毒素实验室能力验证的现状,详细介绍了在真菌毒素能力验证中使用的检测技术研究进展。以Z比分数法评价由比利时联邦药物和农业化学研究中心(CODA-CERVA)组织的粮油多种真菌毒素含量测定的国际比对结果,分析粮油样品中多种真菌毒素的测定情况。 相似文献
4.
5.
《中国油料作物学报》2016,(2)
为花生质量安全风险监测和风险评估提供参考,依据食品风险期望值排序方法,收集并检测604个来自8个省市的花生样本,按照花生真菌毒素检测数据和限量标准,对花生中真菌毒素风险因子危害程度进行标识,确立花生真菌毒素风险因子评价指标,建立了一种花生中真菌毒素危害风险因子查找、识别和排序的规范方法,并给出了花生中真菌霉毒素危害风险因子风险排序实例。 相似文献
6.
【目的】在前期克隆了一个水稻小热休克蛋白(OsHSP20)的基础上,进一步分析该蛋白多克隆抗体的免疫反应特性和应用范围,为深入鉴定该类基因的功能奠定基础。【方法】利用合成多肽和其原核表达产物分别免疫家兔制备了相应的多克隆抗体;ACP(Antigen-coated plate)-ELISA和dot-ELISA用于分析比较多克隆抗体效价和灵敏度;Western blot用于鉴定多克隆抗体的特异性和应用范围。【结果】Western blot分析显示,利用重组OsHSP20蛋白及其合成多肽所制备的多克隆抗体均可以在原核表达OsSHSP样品中特异性检测到1条大小与预期一致(37kD)的条带,表明两种方法获得的多克隆抗体都具有高度的特异性;ACP(Antigen-coated plate)-ELISA和dot-ELISA分析表明,利用重组蛋白制备的多克隆抗体效价和灵敏度均高于利用多肽制备的抗体。进一步的Westernblot分析显示该抗体可用于多种植物小热休克蛋白(SHSP)的检测。【结论】制备了OsHSP20蛋白质抗体,且具有高度的特异性和较高的效价和灵敏度,可广泛地用于多种单、双子叶植物HSP20蛋白的表达分析,有助于OsHSP20及其他植物同源蛋白的功能鉴定。 相似文献
7.
为剧毒靶标(黄曲霉毒素)绿色分析提供高效抗体,在已有抗黄曲霉毒素杂交瘤细胞株1Cll的基础上,通过小鼠腹水法制备单克隆抗体,经胃蛋白酶酶解,制备F(ab')2片段,发现在优化条件37C、pH4.1柠檬酸缓冲液中酶解4.5h,可高效制备F(ab')2片段.通过酶联免疫吸附法(ELISA)比较了抗体片段与原始抗体的识别活性,发现F(ab')2片段效价达到1∶320000,是原抗体效价的1.5倍;灵敏度(IC50)为8.7pg/mL,保持了原抗体的抗原结合能力. 相似文献
8.
污染粮油食品的主要真菌毒素及胶体金免疫层析技术在快速检测中的应用 总被引：3，自引：1，他引：2
污染粮油食品的真菌毒素主要有黄曲霉毒素、T-2毒素、玉米赤霉烯酮、脱氧雪腐镰刀菌烯醇和伏马菌素B1。胶体金免疫层析技术是20世纪90年代初建立的一种简易、快速的免疫学检测技术,现已广泛应用于残留检测、生物医学等许多领域。本文介绍了污染粮油食品的主要真菌毒素及胶体金免疫层析技术在粮油食品真菌毒素快速检测中的应用,并对其发展前景进行了展望。 相似文献
9.
10.
采集吉林省主产区玉米280份,采用液相色谱串联质谱法测定13种真菌毒素含量,研究吉林省玉米中多种真菌毒素污染和膳食暴露风险状况,评价膳食暴露风险。结果表明,供试玉米中真菌毒素的平均含量为522.3μg/kg,真菌毒素污染种类分别是伏马菌素(97.9%)、脱氧雪腐镰刀菌烯醇(52.1%)和玉米赤霉烯酮(37.5%),平均污染水平分别为1269.1、284.2、42.8μg/kg。采用残留平均值计算成人和儿童经玉米摄入的伏马毒素、脱氧雪腐镰刀菌烯醇及玉米赤霉烯酮,结果无显著慢性摄入风险。以最大残留值计算摄入呕吐毒素的膳食暴露风险,成人和儿童无显著急性摄入风险。吉林省玉米中真菌毒素的污染呈多种类污染普遍的现象,居民通过玉米摄入真菌毒素的风险熵值很小,对人体健康潜在风险不大。 相似文献
11.
量子点标记荧光免疫法检测花生中黄曲霉毒素B1 总被引：2，自引：0，他引：2
建立了基于量子点标记二抗的间接竞争荧光免疫吸附测定方法（indirect competitive fluorescence-linked immunosorbent assay,cFLISA）,并研究检测花生中黄曲霉毒素B1可行性。采用谷胱甘肽为稳定剂,在水相中直接合成碲化镉（CdTe）量子点,并利用EDC与兔抗鼠二抗进行共价偶联,以黄曲霉毒素B1的单克隆抗体建立cFLISA方法。结果表明,该方法的灵敏度和最低检测限值分别为0.023ng/mL和0.001ng/mL,与传统的有机染料FITC-二抗法比较,灵敏度提高了30倍,花生样品加标0.1、0.05和0.025ng/g,回收率范围在88%~116%之间,变异系数均小于10%。建立的cFLISA方法可以较好的检测花生中黄曲霉毒素B1,并为其它真菌毒素的检测提供参考。 相似文献
12.
The present study aimed to identify mycotoxins in edible tissues of Atlantic salmon (Salmo salar) using liquid chromatography coupled to hybrid quadrupole time-of-flight mass spectrometry (LC-Q-TOF-MS). After using a non-targeted screening approach and a home-made spectral library, 233 mycotoxins were analyzed. Moreover, the occurrence of mycotoxins in fish filets was evaluated, and their potential toxicity was predicted by in silico methods. According to the obtained results, forty mycotoxins were identified in analyzed salmon samples, the predominant mycotoxins being enniatins (also rugulosin and 17 ophiobolins), commonly found in cereals and their by-products. Thus, mycotoxin carry-over can occur from feed to organs and edible tissues of cultivated fish. Moreover, the toxicity of detected mycotoxins was predicted by the in silico webserver ProTox-II, highlighting that special attention must be paid to some less reported mycotoxins due to their toxic predicted properties. 相似文献
13.
People affected by coeliac disease need to adhere to a life-long gluten-free diet to avoid symptoms. ELISA-tests are seen as the mainstay for the detection of gluten in gluten-free food because of their sensitivity. They can, however, yield different gluten amounts depending on the antibody and reference material used. We compared the reactivity of three prominent mouse anti-gliadin-antibodies to a reference gliadin isolated from 28 common bred European wheat varieties. The reference material proteins were labelled with fluorescent dye Cy3. They were then separated by 2DE and transferred by Western blot onto low fluorescent PVDF-membranes, followed by incubation with the three primary anti-gliadin antibodies one by one. Detection of the reacting proteins used anti-mouse antibody which was labelled with fluorescent dye Cy5. The use of this technique made it possible to co-detect the 2DE-image of the reference material proteins (Cy3) and proteins reacting with the respective antibody (Cy5). The three investigated antibodies had dissimilar reactivities with different proteins of the reference gliadin. Antibodies R5 and PN3 reacted mainly with gliadin fractions, antibody 401.21 mainly with high molecular weight glutenins. The results confirm the individual specificity of these antibodies and demonstrate the importance of validating immunochemical methods for gluten detection. 相似文献
14.
我们于1988年建立了以水稻白叶枯病菌单克隆抗体为第二抗体的酶联免疫吸附测定法,虽然具有灵敏、精确、简便、快速的优点,但用于检测水稻白叶枯病病种所带的微量细菌,灵敏度仍然不足,存在漏检现象。因而,我们对利用单克隆抗体检测水稻白叶枯病病种的方法作了进一步的改进研究。 相似文献
15.
Silage and total mixed ration hygienic quality on commercial farms: implications for animal production 
下载免费PDF全文
下载免费PDF全文 T. Cogan R. Hawkey E. Higgie M. R. F. Lee E. Mee D. Parfitt J. Raj S. Roderick N. Walker P. Ward J. M. Wilkinson 《Grass and Forage Science》2017,72(4):601-613
Implications of silage hygienic quality for animal production were investigated on forty‐five dairy farms in South West England. Samples of grass and maize silages and of total mixed rations (TMR) were obtained together with information on silage technology, herd size and animal production. Samples were analysed for mycotoxins, bacteria, yeasts, moulds and chemical composition. Thirteen mycotoxins were assayed, but none were detected in the samples of grass silage. However, mycotoxins were found in 0·9 of all maize and other silage samples, with deoxynivalenol and zearalenone predominating. There was no relationship between total mycotoxin concentration and mean lactation milk yield per cow. Enterobacteria counts tended to be higher in maize silage than in grass silage and higher still in TMR – a cause for concern. There were no relationships between mould counts and mycotoxin concentrations in silages, implying that mycotoxins may have been produced in the field pre‐ensiling. 相似文献
16.
Toxicogenic fungal moulds have been previously identified in Irish farm silages and are known to be capable of producing mycotoxins. Mycotoxins are detrimental to animal health and performance and can be transferred through feed to animal produce. The objectives of this study were to identify and quantify the challenge posed to livestock from mycotoxins in Irish silages and determine whether conventional chemical characteristics could be used as indicators of mycotoxin occurrence. Over the 2‐year period, 300 silages were sampled on 150 farms and these consisted of round‐baled grass silages (n = 115) and pit silages of either grass (n = 175) or maize (n = 10). There was no significant difference in mycotoxin concentrations across silage types in either year, except for baled silage containing higher concentrations of enniatin B compared with pit silage in Year 1. Conventional chemical characteristics of silages were generally not reliable predictors for mycotoxin incidences; however, dry‐matter digestibility, crude protein, fermentation products and ash did predict the incidence of enniatins and beauvericin. The incidence and concentration of the twenty measured mycotoxins were generally low and individual mycotoxin concentrations recorded were considerably lower than current EU directive or guidance thresholds. Non‐regulated mycotoxins measured were similar to or lower than concentrations reported in the literature. Based on current knowledge and the concentrations of individual mycotoxins detected in this study, the challenge to Irish livestock and livestock products from animal consuming silage is generally low. However, the additive or synergistic effects of multiple mycotoxins in silage are unknown. 相似文献
17.
Antibody molecules, and antibody fragments in particular, have enormous potential in the development of biosensors for marine monitoring. Conventional immobilisation approaches used in immunoassays typically yield unstable and mostly incorrectly oriented antibodies, however, resulting in reduced detection sensitivities for already low concentration analytes. The 2H12 anti-domoic acid scFv antibody fragment was engineered with cysteine-containing linkers of two different lengths, distal to the antigen binding pocket, for covalent and correctly oriented immobilisation of the scFvs on functionalised solid supports. The Escherichia coli-produced, cysteine-engineered scFvs dimerised in solution and demonstrated similar efficiencies of covalent immobilisation on maleimide-activated plates and minimal non-covalent attachment. The covalently attached scFvs exhibited negligible leaching from the support under acidic conditions that removed almost 50% of the adsorbed wildtype fragment, and IC50s for domoic acid of 270 and 297 ng/mL compared with 1126 and 1482 ng/mL, respectively, for their non-covalently adsorbed counterparts. The expression and immobilisation approach will facilitate the development of stable, reusable biosensors with increased stability and detection sensitivity for marine neurotoxins. 相似文献