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转基因食品检测技术及方法不断发展和完善
发布时间: 2014-06-20 - 标签: , - 浏览 1,814 次

所谓转基因食品,就是通过基因工程技术将一种或几种外源性基因转移到某种特定的生物体中,并使其有效地表达出相应的产物(多肽或蛋白质),此过程叫转基因。现在社会上对转基因食品的安全度关注点极高,所以对转基因食品的检测技术也相应地在不断发展和完善。

根据转基因食品来源的不同可分为植物性转基因食品、转基因酵母疫苗、转基因工程菌抗生素、动物性转基因食品和微生物性转基因食品。目前,我国的不少转基因因技术属世界领先水平,但应用很少。

  转基因食品的检测技术

由于转基因物质有可能在耕种、收割、运输、储存和加工过程中混入食品,对食品造成偶然污染。因此,不论是对转基因食品贴示标签,或是对转基因与非转基因原料进行分别输送,转基因原料和旧食品的检测都是必不可少的;另外,要区分转基因与非转基因食品,对转基因食品进行选择性标记,对食品中的转基因含量的多少加以限制,也需要准确有效的基因检测技术。

目前,转基因存在的检测方法主要有prc(凝合酶链反应)技术、凝胶电泳测序和elisa(酶联免疫法)等,这些方法在食品和医药研究方面都有广泛的使用,其中以prc技术最为成熟。该技术是由美国Centus公司的Karymullis发明,于1985年由saiki等首次报道,是近年来开发的体外快速扩增dna技术。1988年,r.k.saiki等人又成功地将热稳定taqdna聚合酶应用于pcr扩增,是pcr技术上向实用化的一次突破性的进展。通过pcr技术可以简便快速地从微量生物材料中以体外扩增的方式获得大量特定的核酸,并且有很高的灵敏度和特异性。pcr既可做定性又可做定量分析,目前大多以定性检测为主,定性检测的检出范围为百分之0.1。但该项技术的检测结果也有可能与实际不相符合,会出现假阴性或假阳性结果(即检测物质本身含有转基因物质,而未被检出;或是本身没有转基因物质,而检出有转基因成分)。

由于许多获得批准的遗传工程体(gmo)表现出一定的导入基因的性状,因此,基因检测也需要以蛋白质为基础的检测技术,这就出现了elisa法。1971年Engvall和Perlman发表了有关酶联免疫吸附分析法用于免疫球蛋白g(igg)定量测定的文章,使该技术发展成为液体标本中微量物质的测定方法。elisa法与pcr法相比具有操作简单、快速、结果准确,有高通量的特性,解决了转基因检测中样品核酸制备的困难,同时降低了检测成本和时间高的催化效率,可极大地放大反应效果,从而使测定方法达到很高的灵敏度和稳定性。但该项技术主要应用于原料和半成品分析,在终产物分析方面灵敏度底于pcr法。另外,该项技术还需要特殊的抗体和“新”的表达蛋白,而食品中的这类“新”蛋白质的含量极低,且在加工过程中蛋白质常会发生变化;该分析法同时还需要熟练的操作技术。这些因素都使elisa法在转基因检测应用上受到了一定的限制。

  转基因食品的检测方法

前转基因的检测方法转基因作物检测方法大体分为两种:一是检测是否含有外源蛋白,即外源基因表达产物,主要采用酶联免疫吸附法和试纸条法;二是检测是否含有外源基因(DNA),主要有Southern杂交技术、基因芯片法和PCR检测法。理论上来说,以上所介绍的转基因检测方法,只要改变相应的检测靶标,都可检测不同的转基因产品。

但是实际应用中,以检测外源蛋白的转基因检测方法存大很大的局限性,食品的复杂成分会干扰检测效果,并且加工过的转基因食品中的蛋白质抗原性很容易受破坏,从而会影响检测结果的灵敏度。此外,该方法需要大量的抗体和抗原。目前,商品化的ELISA检测试剂盒和胶体金试纸条只能检测少数几种转基因产品,且一种试剂盒或试纸条只能针对某一特定转基因产物,不能针对多种混合成分的食品样品进行检测。

基因水平的检测方法具有适用范围广的优势,只要样品中有DNA存在,就能被检测,不受食品混合、复杂成分的干扰,适用于转基因原料、初加工和深加工食品的检测,而且有很好的特异性和灵敏度,已经发展为常用的转基因食品检测方法。目前,所有商业化的转基因食品都有相应的PCR检测方法标准。目前,转基因PCR检测产品多数产自国外,价格都比较高,国内杭州有个化工仪器网的平台,转基因检测产品在算是国内最齐全的,有仪器及配套试剂。

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