转基因番木瓜及其深加工制品的PCR检测方法

对番木瓜不同材料及其深加工制品的核酸提取方法进行了优化。成功提取出DNA。选取番木瓜特异性的内源蛋白PAPAIN基因片段作为内参基因，用于检验提取DNA的质量，有效避免了聚合酶链式反应PCR的假阴性并能用于确定检测样品中是否含有番木瓜成分。选取国内目前应用比较多的外源目的基因环斑病毒外壳蛋白（CP）基因以及复制酶（RP）基因作为品系鉴定的基因。适用于出入境检验检疫部门和农业部门对转基因番木瓜及其制品的检验监测。     

采用多重PCR技术检测转基因番木瓜华农一号

为了建立针对转基因番木瓜华农一号简便、快速、高灵敏度的多重PCR技术,用DP305植物基因组试剂盒方法提取3种转基因番木瓜(Event 55-1、GM-YK和华农一号)和非转基因番木瓜(台农2号)样品DNA,以木瓜蛋白酶基因(Papain)作为内参基因,进行了单重PCR技术引物特异性验证,多重PCR方法条件摸索及引物的灵敏度确定。结果表明∶5对特异性引物(CP、35s、NPTII、Papain、RP)得到相应的不同大小单一条带,其大小分别103、166、213、281、369 bp,说明引物具有特异性;建立了检测转基因番木瓜华农一号的五重PCR检测技术,检测限为0.2 ng。     

2012~2015年深圳市售转基因大豆及其产品的监测结果分析

目的 了解2012~2015年深圳市售转基因大豆及其制品的市场占有率、标识情况及发展态势。方法 在深圳各大连锁超市随机抽取大豆及其制品, 采用试剂盒法提取样品DNA, 采用实时荧光PCR的方法扩增大豆内源基因Lectin, 并扩增外源基因CaMV35S启动子和NOS终止子对样品进行定性筛查, 分析不同类别、不同年份样品的阳性率差异及变化趋势。结果 4年监测大豆及豆制品共283份, 检出阳性样品19份,总体阳性率为6.71%; 大豆样品的转基因阳性率为0; 不同年份的豆制品转基因阳性率之间存在显著性差异且有逐年上升的趋势; 所有检出的阳性样品均未按照规定标识。结论 目前深圳市场转基因大豆及其制品(大豆油除外)的市场占有率还很低, 但豆制品转基因阳性率有逐年上升的趋势。政府应该加强转基因产品的标识管理, 并加强转基因农产品的监管。     

2011—2012年北京市丰台区市售食品中镉污染状况分析

了解北京市丰台区部分市售食品中镉的污染状况。方法 采取分层随机抽样方法,2011—2012年间采集丰台区大型农副产品批发市场、商场超市、农贸市场共6类215份市售食品样品,按照GB/T 5009.15—2003《食品中镉的测定方法》对样品进行前处理,采用石墨炉原子吸收光谱法检测镉含量。结果 2011—2012年监测样品中镉的检出率为76.7%(165/215);按照GB 2762—2012规定的限量值,以及根据鲜香菇的含水量进行折算后的干香菇的限量值进行比较,则2011—2012年监测样品的合格率为100%。结论 丰台区各类市售食品镉污染水平总体是安全的,但各类市售食品都有检出镉,说明存在不同程度的污染,需要加强监测,控制污染,保障人群的健康。     

转基因食品在规范中求发展

20世纪80年代以来,以转基因技术(GeneticallyModified Technology)为核心的现代生物技术产业开始蓬勃发展。它为解决人口膨胀、粮食短缺、资源匮乏、环境污染、能源危机等问题提供了崭新的解决之道。但是目前国际社会并没有普遍承认转基因技术,许多人把它看成了一把“双刃剑”,认为转基因     

潮州市市售豆制品的转基因成分检测

为了解潮州市区市售豆制品的转基因成分状况,随机购买潮州市区主要市场市售豆制品进行检测与分析。用CTAB法即改良十六烷基三甲基溴化铵,对豆制品的DNA成分进行提取并对提取到的豆制品DNA用核酸蛋白分析仪测量OD值,利用核酸定性聚合酶链式反应(Polymerase Chain Reaction,PCR)、实时定量PCR等技术对外源基因CP4-EPSPS、NOS和Ca MV35S进行检测。对潮州市区主要市场市售豆制品29份样品进行检测,研究结果表明,提取到的豆制品DNA溶液浓度和纯度都较高,OD260 nm/OD280 nm介于1.9～2.1之间,抽取到的DNA质量浓度达到200 ng/μL以上,能够满足检测要求。在检测的豆制品中,有超过一半含有转基因成分。除了大润发超市外,豆制品没有任何食品标签,所有豆制品也没有是否含有转基因成分的标签。转基因食品标识制度的实施仍处于一个初始阶段,国家应该建立起强有力的市场监管机制。因此,我们必须高度重视转基因食品标签制度落实及其相应的市场监管工作。     

番木瓜及其深加工制品中转基因成分定性PCR检测方法的建立(英文)

本研究对番木瓜不同材料及其深加工制品的核酸提取方法进行了优化,成功提取出可用于PCR扩增反应的DNA。选取番木瓜特异性的内源蛋白PAPAIN基因片段作为内参基因,用于检验提取DNA的质量,有效避免了PCR反应的假阴性并能用于确定检测样品中是否含有番木瓜成分。选取国内目前应用比较多的外源目的基因环斑病毒外壳蛋白(CP)基因以及复制酶(RP)基因作为品系鉴定的基因。本方法适用于出入境检验检疫部门和农业部门对转基因番木瓜及其制品的检验监测。     

复合PCR荧光检测方法检测转基因水稻品系

建立一种高通量的转基因复合聚合酶链式反应（polymerase chain reaction,PCR）荧光检测方法,该方法通过对转基因检测内源、外源及品系基因检测引物进行荧光标记,经复合PCR扩增和毛细管电泳荧光检测,实现特异性强、高通量检测。结果表明,复合PCR荧光检测方法检测4 种转基因水稻品系特异性好,并且灵敏度达到0.1%,可以在进出口检验检疫、食品安全监测等领域进行应用。     

转基因生物食品安全检测分析

综合运用当前国际上先进的基于核酸水平的外源转基因成分检测技术,通过DNA提取、Southern杂交试验、转基因结构的检测、外源基因拷贝数的检测等4项试验,从分子水平和生产角度上对转基因肉鸡进行了一次完整的安全检测分析,这一整套方法对生物食品的生产安全性进行了检测,取得了一次的成效。     

Detection of recombinant DNA from genetically modified papaya

A method using polymerase chain reaction (PCR) was developed to detect the genetically modified (GM) papaya (55-1 line), of which the mandatory safety assessment has not been finished in Japan because of insufficient data. The papaya intrinsic papain gene was used as an internal control. The results of PCR amplification of the papain gene segment indicated that a commercial silica membrane type kit (QIAGEN DNeasy plant mini) was useful for extraction of DNA from papaya fruit, but not for extraction from canned papaya fruit. On the other hand, a commercial ion-exchange type kit (QIAGEN Genomic-tip) provided enough purified DNA for PCR from canned papaya fruit. Compared with the parental line and other commercial non-GM papayas, the DNA from GM papaya fruit provided specific amplification bands in PCR with five primer pairs (Nos. 2-6) including beta-glucuronidase and neomycin phosphotransferase II gene-specific ones. On the other hand, the primer pairs recognizing these genes showed false-positive results when we used DNAs from canned papaya. Therefore, we recommend that the primer pairs (Nos. 5 and 6) recognizing the sequences derived from two different species of organism should be used in order to detect specifically the GM papaya in canned fruits.     

Detection method for genetically modified papaya using duplex PCR

A simple and rapid method for the identification of genetically modified (GM) papaya, derived from Line 55-1, was developed by modifying the Japanese official PCR method. Genomic DNA was directly extracted from the fresh fruit without the lyophilization step, using a commercial silica-based kit. To develop a duplex PCR method which simultaneously detects the GM papaya-specific gene and the intrinsic papain gene, the papain 2-5'/3' (amplicon size; 184 bp) primer pair for the detection of the papain gene was newly designed within the region of the products (211 bp) amplified using the papain 1-5'/-3' primer pair adopted in the Japanese official PCR method. To detect the GM papaya-specific gene, the primer pair Nos C-5'/CaM N-3' described in the Japanese official method was used. The DNA sequences of the GM papaya gene and the intrinsic papain gene were co-amplified using the PCR method in a single tube. The developed duplex PCR method allows the simultaneous detection of the products by means of agarose gel electrophoresis or microchip electrophoresis. The proposed method for GM papaya identification is simple and rapid.     

转基因棉的检测研究

转基因棉在全球范围内的种植量已有很大程度的增加,同时对有机棉的需求也在不断增长。所谓“有机”,就是禁止基因工程。随之而来的是不可忽视的污染和冒牌问题。针对于这些问题介绍了一项关于转基因棉纤维检测的十分可靠灵敏的分析程序。     

Detection of genetically modified organisms in processed meat products on the Serbian food market

The addition of soybean proteins to processed meat products has significantly increased in recent years due to the interesting functional and nutritional properties of these vegetable proteins. Since the Roundup Ready (RR) soybean is the only transgenic soybean line approved for market in EU this work was aimed at monitoring its presence in meat products on the Serbian food market. The extracted DNA was analyzed using duplex polymerase chain reaction (PCR) with primer pairs aimed at the lectin gene and 35S promoter. Samples positive for the presence of GM soybean were subjected to a real-time quantification of the percentage of RR soya. The results indicated that out of fifty processed meat products examined, twelve gave positive results with 35S promoter and all contained RR soya below 0.1%.     

2011年深圳转基因大豆及其制品的市场占有率和标识情况调查

目的 了解深圳市场转基因大豆及其制品的市场占有率和标识情况,为政府监管转基因大豆提供科学依据.方法 按照监测计划,定期在深圳市场随机抽取大豆及其制品,采用实时荧光PCR方法对其进行定性和定量检测,评估深圳市场转基因大豆的市场占有率和标识情况,并与2006年的监测结果进行比较和分析.结果 2011年共监测样品106份,检出2份转基因阳性样品,市场占有率为1.89％;两份阳性样品中转基因大豆的含量分别为0.93％和0.51％,定量检出限为0.1％.所有检出转基因阳性样品均没有转基因标识;与2006年结果比较,转基因大豆及其制品的市场占有率以及标识情况差异均无统计学意义.结论 目前深圳市场转基因大豆及其制品(食用油脂除外)的市场占有率还很低,并且5年来未发现明显上升;转基因产品标识的管理还有待加强,政府应该加大这方面的执法力度.     

进口薯格中转基因成分的检测

目的建立了一种快速检测进出口食品中转基因成分的方法。方法本实验采用DNA提取试剂盒对薯格中的DNA进行快速提取,接着用实时荧光PCR方法对其进行转基因成分和品系的鉴定。结果通过对食品标签进行初筛,发现其标识成分中含有未标明的转基因成分。进一步对所含转基因成分的品系进行鉴定,确定薯格的外包裹玉米粉中含有多种转基因成分,包括转基因玉米TC1507、NK603、MON810、59122、MON89034等5个品系。结论本文方法可以用于加工食品中转基因玉米成分及品系的定性检测,也可以作为常规PCR定性方法的确证试验方法。     

Detection approaches for genetically modified organisms in foods

This review examines the various detection strategies for genetically modified organisms (GMOs) in food products. It begins with a brief discussion of the issues related to the technology especially the risks and public concerns. An introduction to the biological aspects of the major GMOs then follows. The bulk of the review is concerned with the different approaches toward detection: (a) PCR-based methods such as real-time, duplex and multiplex, (b) the use of biosensors and microarrays, (c) the presence of commercially available kits, and (d) other methods such as electrophoresis and wavelength-dispersive X-ray fluorescence. Each of these methods is critically discussed and various applications are described.     

食用大豆油中转基因成分的检测

食用油脂中转基因成分的检测是当前食品检验工作的一个主要方面。针对食用油脂中DNA含量极低、DNA序列片段短、破坏严重的特点,建立了食用油脂中DNA提取方法,通过实时荧光PCR可检测出大豆内源基因(Lectin)以及外源抗除草剂基因EPSPS,为食用油脂进行核酸类生物性检测提供了一种简捷有效的方法。     

A histochemical method using a substrate of beta-glucuronidase for detection of genetically modified papaya

A histochemical assay for detecting genetically modified (GM) papaya (derived from Line 55-1) is described. GM papaya, currently undergoing a safety assessment in Japan, was developed using a construct that included a beta-glucuronidase (GUS) reporter gene linked to a virus coat protein (CP) gene. Histochemical assay was used to visualize the blue GUS reaction product from transgenic seed embryos. Twelve embryos per fruit were extracted from the papaya seeds using a surgical knife. The embryos were incubated with the substrate 5-bromo-4-chloro-3-indolyl-beta-D-glucuronide (X-Gluc) in a 96-well microtiter plate for 10-15 hours at 37 degrees C. Seventy-five percent of GM papaya embryos should turn blue theoretically. The histochemical assay results were completely consistent with those from a qualitative polymerase chain reaction (PCR) method developed by this laboratory. Furthermore, the method was validated in a five-laboratory study. The method for detection of GM papaya is rapid and simple, and does not require use of specialized equipment.