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动物产品中牛、羊源性成分多重PCR检测方法的建立 总被引:14,自引:2,他引:14
以肉骨粉、鱼粉、猪肉干和鱼肉干为研究对象,异硫氰酸胍法提取总DNA,18S rDNA片段的扩增结果表明提取到的DNA中不存在抑制PCR的物质。应用梯度PCR技术对牛、羊源性成分检测的退火温度进行了优化,在单一PCR检测技术的基础上分别进行了18S rDNA片段和牛、羊源性成分的多重PCR分析,得到了预期的结果。试验表明,本文建立的多重PCR方法具有快速、简便、准确等特点,对动物产品牛、羊源性成分检测具有重要意义。 相似文献
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曹志玲 《养殖与饲料.饲料世界》2010,(5):49-51
流行病学调查证明,用患有疯牛病和羊痒病的牛、羊等动物的肉和骨头制成的肉骨粉喂养动物会导致疯牛病和羊痒病的传播,是引起疯牛病发生的最初原因。目前,用于检测饲料中牛、羊源性成分的主要方法有:以检测骨头为基础的显微镜方法;以检测蛋白质为基础的免疫学方法和以检测DNA为基础的检测方法;其它方法还包括近红外光谱法等。 相似文献
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根据牛、羊线粒体中物种特异性序列设计引物,应用可视化环状等温扩增技术(visual loop-mediate isothermal amplification,vLAMP)对牛、羊源性成分进行快速扩增及可视化判断,同时建立牛、羊源成分人工污染的饲料模型,考察vLAMP检测方法的灵敏度。结果表明,牛源性引物和羊源性引物仅分别对牛、羊源性成分产生特异性扩增,灵敏度高达0.001%。本试验所建立的牛、羊源性成分现场可视化筛查方法具有较高的特异性与灵敏度,操作简单、快速,可用于饲料中牛、羊源性成分污染和食品掺伪的现场可视化检测。 相似文献
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为鉴定和区分饲料及动物产品中牛、山羊、绵羊源性成分,根据线粒体DNA(mitochondrial DNA,mtDNA)种间保守序列,设计合成了3对特异性引物与TaqMan探针,通过对荧光PCR反应体系和反应条件的优化筛选,建立了三重荧光PCR方法,在同一个荧光PCR反应中完成3种动物源性成分的检测。用该方法对16种不同源性的动物DNA进行检测,结果表明能特异地鉴别检测出牛、山羊和绵羊源性成分,且敏感性比现行国标PCR法高100倍。该方法适用于饲料、肉制品、奶制品等动物源性产品的检测。 相似文献
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Immunosuppressive activity of conditioned medium from cultured ovine, caprine, and hybrid trophoblast tissue was examined. Conceptuses were obtained from naturally mated donor ewes and does at d 20 of gestation and trophoblast tissue was cultured for 24 h in medium supplemented with 15% calf serum and 1% antibiotic/antimycotic. Conditioned medium was added to pokeweed mitogen-stimulated sheep and goat lymphocyte cultures. Quantification of [3H]thymidine uptake by cells was used to measure lymphocyte proliferation. Ovine, caprine, and hybrid conditioned medium effectively suppressed sheep and goat lymphocyte proliferation (P less than .01). There were no differences (P greater than .05) between the immunosuppressive activity of the three tissue types on either sheep or goat lymphocytes. For all treatment groups, sheep lymphocytes were suppressed more than goat lymphocytes (P less than .05). These results indicate that, at d 20 of gestation, sheep x goat hybrid trophoblast tissue is capable of suppressing pokeweed mitogen-stimulated lymphocyte proliferation. 相似文献
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根据线粒体基因的特异性鉴别检测饲料中的动物源性成分 总被引:8,自引:1,他引:8
为了防止海绵状脑病疫区和痒病疫区的反刍动物源性饲料入手动物食物链,继而进入人类消费的食物链,因而,非常用必要对贸易往来中的肉骨粉品种来源进行鉴定检测。我们运用聚合酶链反应技术(Polymerase Chain Reaction,PCR),以检测不同种动物特异性线粒体基因片段为目标,分别建立了从动物源性饲料中鉴别检测猪、鸡、马成分的方法。该方法具有灵敏度高、快速和特异性强的优点,可作为动物源性饲料中猪、鸡、马源性成份鉴别检测的常规方法之一。 相似文献
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E E Brako R W Fulton S S Nicholson G F Amborski 《American journal of veterinary research》1984,45(4):813-816
Sera from healthy sheep were collected in January and March 1982 from flocks of sheep located in southwestern and southeastern Louisiana. These sera were tested for bovine herpesvirus-1 (BHV-1), bovine viral diarrhea virus (BVDV), parainfluenza-3 (PI-3) virus, and goat respiratory syncytial virus (GRSV) antibodies by microtitration virus-neutralization test. The sera were tested also for bovine leukemia virus (BLV) and bluetongue virus (BTV) antibodies by immunodiffusion tests. The number of flocks with seropositive sheep for each virus were: 2/8 (25%) for BVDV; 8/8 (100%) for PI-3 virus; 7/8 (87.5%) for GRSV; and 6/8 (75%) for BTV. Seropositive rates for each virus for the individual sheep tested were: 4/158 (2.5%) for BVDV; 117/158 (74.1%) for PI-3 virus; 77/158 (48.7%) for GRSV; and 21/158 (13.3%) for BTV. All sheep were seronegative for BHV-1 and BLV. 相似文献
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利用多重PCR检测兔肠致病性大肠杆菌和魏氏梭菌的毒力基因 总被引:3,自引:0,他引:3
我们建立了一种同时检测兔肠致病性大肠杆菌和魏氏梭菌毒力基因的二重PCR方法。在一次PCR反应中同时扩增兔肠致病性大肠杆菌(REPEC)的eae毒力基因和魏氏梭菌的α-toxin毒力基因,扩增产物经电泳,出现与阳性对照细菌条带大小一致的条带即判断为该标本阳性,本试验还对79份腹泻兔粪便或肠内容物样品进行了检测,结果35.4%(28/79)检出eae+的REPEC,而魏氏梭茵的检出率为。6.3%。敏感性试验结果表明,该二重PCR能检出10 CFU的REPEC,10 CFU的魏氏梭菌,具有简便、快速、特异、经济等特点,宜于临床应用。 相似文献
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根据鱼线粒体DNA(mtDNA)基因组序列,选择高度保守区域设计特异性引物和TaqMan探针,通过反应条件优化,建立了饲料中鱼源性成分检测的实时荧光PCR方法.结果显示,应用该方法只能检测到鱼源性成分,表明该方法具有良好的特异性,对饲料中鱼源性成分的最低检出限为0.05%,定量检测的线性范围0.018~18.46ng.试验结果表明,该方法能对饲料中鱼源性成分进行准确、快速的检测,具有特异性好、灵敏度高的优点,对防止饲料掺假、控制进出口饲料安全具有重要意义. 相似文献