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食品中鲨鱼源性成分真实性PCR鉴别研究 总被引:7,自引:0,他引:7
建立食品中真实性鲨鱼源性成分PCR鉴别方法,该方法特异性强,灵敏度高。运用普通PCR技术对9种鲨鱼及42种常见非鲨鱼类动植物样品进行检测,9种鲨鱼中出现180bp的特异性扩增条带,其他非鲨鱼样品中均未出现扩增条带,实验表明,本检测方法具有特异性,检测限为0.1ng/μL鲨鱼DNA和0.1%(W/W)鲨鱼肉粉。运用建立的方法对20种常见的鲨鱼产品进行PCR检测,除仿鱼翅和鲨鱼肝油外,所有产品中均能检测出鲨鱼成分。该检测方法能够用于食品中鲨鱼源性成分的真实性鉴别。 相似文献
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根据狐狸线粒体基因组中的保守序列和貉子线粒体D-loop基因保守序列设计狐狸、貉子特异性引物和Taq Man探针,建立一种基于实时荧光聚合酶链式反应的肉及肉制品狐狸、貉子源性成分测定方法,通过特异性、灵敏性、线性检测对该方法体系进行检验和评价。本研究建立的狐狸、貉子源性成分实时荧光聚合酶链式测定方法体系具有良好的特异性及灵敏性,最低可检测0.5 pg/μL纯狐狸DNA和5 pg/μL纯貉子DNA。本研究建立的狐狸貉子源性成分荧光PCR检测方法,可以用来检测实际样品中是否含有狐狸貉子源性。 相似文献
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依据牛种属的线粒体细胞色素b(cytochrome b,Cytb)基因上的保守序列设计牛源特异性引物对,建立一种特异性强、灵敏性高、耗时短的鉴定食品中牛源性成分的聚合酶链式反应(polymerase chain reaction,PCR)方法。选取牦牛、水牛、黄牛品种及羊、猪、鸡、鸭、鹅、兔、马、驴、鱼、大豆、玉米等动植物源成分进行特异性试验。结果显示:所建立的方法高度特异于牛源性检测,其他非牛动植物源成分均无扩增条带。采用常见的牛品种进行灵敏性试验,结果显示所建立的方法可检测到的最低牛DNA量为0. 2 pg;将牛肉分别与猪肉、驴肉、羊肉、大豆组织混合进行PCR测定,结果表明最低检测限为鲜样品含0. 01%质量分数的牛源成分,高压121℃、0. 1MPa,20 min处理样品含0. 01%质量分数的牛源成分。对市售食品中牛源性成分检测结果与现行PCR牛源性成分检测标准方法的检测结果 100%一致。 相似文献
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保健品中牛羊源性成分的PCR检测 总被引:6,自引:0,他引:6
本文根据牛、羊线粒体mtDNA中的保守序列,设计了针对牛羊源性成分检测的特异性扩增引物,通过聚合酶链反应技术(Polymerase Chain Reaction,PCR)建立了保健品中牛羊源性成分的快速检测方法。通过内切酶DpnⅡ和SspⅠ可分别对牛羊成分的扩增结果进行进一步验证,该方法对牛源性成分的检测低限为0.05%,对山羊和绵羊源性成分的检测低限分别为0.005%和和0.5%,可作为保健品中牛羊源性成分检测有效方法,也可作为保健品及动物源性产品中成分真伪鉴别的准确、可靠方法。 相似文献
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根据羊线粒体DNA细胞色素b基因序列,设计特异性引物和Taqman探针。通过反应条件的优化筛选,构建标准曲线,建立灵敏、可靠的肉制品中羊源性成分的实时荧光PCR鉴别方法。结果表明,所设计的引物可有效地进行肉制品中羊肉成分的鉴别,特异性强,灵敏度高达16pg/μL;标准曲线扩增效率达98.561%,R2=1,符合《实时荧光定量国际化标准-MIQE指南》要求;本研究建立的方法检测结果与国标GB/T 20190-2006方法结果符合率达100%,且不需电泳、酶切和测序等步骤。通过对市售肉制品样本的鉴别验证了方法的实用价值,为肉制品质量的有效控制提供了新的途径。 相似文献
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荧光定量PCR方法鉴别肉制品中羊源性成分 总被引:1,自引:0,他引:1
根据羊线粒体DNA细胞色素b基因序列,设计特异性引物和Taqman探针。通过反应条件的优化筛选,构建标准曲线,建立灵敏、可靠的肉制品中羊源性成分的实时荧光PCR鉴别方法。结果表明,所设计的引物可有效地进行肉制品中羊肉成分的鉴别,特异性强,灵敏度高达16pg/μL;标准曲线扩增效率达98.561%,R2=1,符合《实时荧光定量国际化标准-MIQE指南》要求;本研究建立的方法检测结果与国标GB/T 20190-2006方法结果符合率达100%,且不需电泳、酶切和测序等步骤。通过对市售肉制品样本的鉴别验证了方法的实用价值,为肉制品质量的有效控制提供了新的途径。 相似文献
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建立一种快速、准确鉴定牛肉制品中牛源性成分并且量化牛肉成分含量的方法。以牛线粒体细胞色素b基因为靶基因,设计出具有特异性引物。选择真核生物核糖体16SrDNA为内参基因,采用实时荧光相对定量法。牛肉质量百分比的对数值与之对应的循环阈值差值△Ct呈良好线性关系。标准曲线回归公式为y=-3.3645x+0.8737,R2=0.9926,扩增效率达98.25%。通过模拟混合样品对标准曲线进行质量评估,证明该方法适用于对牛肉成分的鉴定以及含量的检测,为量化肉制品中牛肉成分研究提供参考意见。 相似文献
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Je-Hung Kuo Hsiu-Wei Tsuei Zih-Ling Jia Che-Yang Lin Yuan-Hsin Chang Bo-Lin Chen Jung Kuan Hsu-Yang Lin Lih-Ching Chiueh Daniel Yang-Chih Shih Hwei-Fang Cheng 《Food Analytical Methods》2018,11(4):992-1000
Mullet roe is a product of high economic value in a number of Asian countries, particularly Taiwan. However, actual mullet roe is commonly adulterated by the addition of other species, such as escolar and oilfish. The purpose of this study was to develop a method to detect the ingredient of mullet roe products. Based on the TaqMan real-time PCR assay, we designed the specific primers-probe set (mullet) that targets the mitochondrial 16S ribosomal RNA gene. Meanwhile, the positive amplification control is designed based on the eukaryotic 18S rRNA gene. The PCR amplicon used to identify fish species in processed roe products is smaller than 200 bp. Method specificity was evaluated by analyzing tissue samples of 29 food fish and 9 puffer fish species. No indications of cross-reactivity toward non-target species were observed. Sensitivity and linearity tests were conducted using five-fold serial dilutions of target DNA from processed mullet roe, and we determined that our proposed method has a sensitivity of 1.2 ng. Further tests on a random survey of commercial fish roe products demonstrated the efficacy of the technique in the detection of mullet DNA. The real-time PCR methods developed in this study could be used to verify the labeling of actual mullet roe products. 相似文献
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为有效鉴别市售鱼翅的真伪,针对鱼翅中鲨鱼源性成分的线粒体基因组的部分序列,利用实时荧光聚合酶链式反应(Polymerase Chain Reaction,PCR)技术,通过特异性引物和探针的设计建立起一种有效的真伪鉴别方法。该方法中对样品脱氧核糖核酸(Deoxyribonucleic acid,DNA)的提取方法、DNA检测的浓度灵敏度、质量灵敏度以及方法的特异性进行研究。结果表明鱼翅样品用试剂盒进行提取效果良好,其检测的DNA浓度灵敏度可达1×10~(-5) ng/μL,质量灵敏度可达0.01%。利用该方法对市场上的购买的47份鱼翅类样品和18份非鱼翅样品进行了检测,43样品检测出鲨鱼成分,包括4份仿鱼翅在内的22份未检出鲨鱼成分。 相似文献
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This study describes the total lipids, protein and fatty acids in the meat, liver, brain and adipose tissue of five male and five female nutria (Myocastor coypus). The animals were reared on a specialized farm in Uruguay. Total lipid content was between 1.41% and 1.84% in males and females. Total cholesterol content was between 70.1 mg and 72.7 mg/100 g of wet tissue. The protein content was between 19.56% and 22.34% in males and females. No significant differences (P > 0.05) were detected between males and females for total lipid, cholesterol or protein. Total, saturated, monounsaturated and polyunsaturated fatty acid values were higher (P < 0.001) in female than in male thigh muscle. Pectoral muscle had more (P < 0.05) saturated and monounsaturated fatty acids in females than in males. Subcutaneous adipose tissue from females had the highest (P < 0.01) total, saturated and monounsaturated but not polyunsaturated fatty acids while abdominal adipose tissue from males had more polyunsaturated fatty. 相似文献
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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。 相似文献
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本文针对鱼翅中的鲨鱼成分进行检测鉴定开发了一种快速灵敏的PCR检测方法,可检测鱼翅类食品中是否存在鲨鱼成分。根据鲨鱼线粒体的细胞色素亚基I基因序列设计了鲨鱼特异性引物,扩增长度为228 bp;为了评价方法的特异性,将设计的引物分别针对22份鱼翅样品DNA和37种其它种类DNA进行PCR检测,结果显示,只在鲨鱼鱼翅中能检测出特异的228 bp条带,其它37种物种中均无条带检出。为了评价方法的灵敏度,将鱼翅DNA中掺入了不同比例土豆DNA的样品采用本方法进行了PCR分析,显示方法可检测灵敏度为0.1%(m/m)。随机抽取45份不同类型的鱼翅样品,检测出22份鲨鱼翅均含鲨鱼成分,而21份仿鱼翅均不含鲨鱼成分而含有植物成分。该样品前处理方法、DNA提取方法以及PCR检测方法可广泛应用于食品中鲨鱼成分检测鉴定。 相似文献
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Growth, carcass yield and proximate composition were determined in intensively reared nutria (Myocastor coypus) fed different levels of protein. Growth, food intake and food utilization efficiency were not significantly affected, within the same sex, in animals receiving 16%, 19% or 22% of protein in the diet. Males consumed the same amount of food as females but had greater live and carcass weights at slaughter age than females. Males had the highest meat yield and females had the highest fat content. Independent of dietary protein level, the pectoral muscles of all animals ranged from 19.2% to 23.6% protein, 1.97% to 2.47% total lipids, and 70.1 to 72.0mg of cholesterol in 100g of tissue. In the thigh muscles, the observed ranges were between 21.4% and 22.9% in proteins, 1.83% and 2.07 in total lipids and 69.9 and 71.0mg of cholesterol in 100g of tissue. The determination of classes of lipids shows more phospholipids contents in animals receiving 16% of protein in diet. 相似文献
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北京油鸡是我国珍贵的地方品种,在自然群体中,部分个体表现出多趾性状,该性状是由于其LMBR1基因的246 bp处发生突变,突变位点的碱基由C突变为A时表现出多趾形状。本实验根据多趾北京油鸡肉基因型与华都艾拔益加肉鸡基因型的不同,通过聚合酶链式反应(polymerase chain reaction,PCR)测序法和限制性酶切片段多态性(restriction fragment length polymorphism,RFLP)技术,确定突变位点的碱基为A或C,从而准确鉴定食品中多趾北京油鸡肉与华都艾拔益加肉鸡的区别。 相似文献