实时荧光PCR检测鸭血中的动物源性成分

目的建立实时荧光PCR检测鸭血中鸭、猪、牛等动物源性成分的方法。方法对重庆市火锅店销售的鸭血进行采样检测。提取鸭血样品DNA,进行实时荧光PCR检测,确定循环阈值,分析样品中鸭、猪、牛源性成分。结果 DNA提取液OD_(260 nm)/OD_(280 nm)值为1.8～2.0,纯度较高; 8个市售鸭血样品中2个样品只检出猪源性成分, 1个样品同时检出鸭源性成分和猪源性成分,研究发现所采集的鸭血存在猪血掺假的可能。结论该方法快捷、可操作性强,适用于鸭血中动物源性成分的测定。     

实时荧光PCR法检测明胶的牛、猪源性成分

为了快速准确鉴别明胶的动物源性,根据线粒体DNA基因COX I(cytochrome coxidase subunit I)的种间多态性差异合成特异性引物及探针,通过实时荧光PCR技术扩增与识别特异性基因片段.运用明胶牛、猪源性成分的荧光PCR检测方法并进行特异性、检出限、重复性实验.结果显示:该方法能够快速准确检测明...     

肉制品中牛源性成分多重实时荧光PCR检测 方法的建立及初步应用

目的建立肉制品中牛源性成分的荧光PCR检测方法。方法根据牛特异性线粒体DNA片段,设计合成两对引物,以生、熟牛肉及超市牛肉加工品为材料,建立肉制品中牛源性成分的多重实时荧光PCR检测方法,并用该法与国标法同时对市售的25份肉制品同时进行检测,通过对其他种类的肉源DNA进行扩增验证方法的特异性;对含有不同比例牛肉成分的DNA样本进行检测确定检出限。结果该方法可成功检测出肉制品中的牛源性成分。在25份肉制品检测中,与国标法检测结果一致。该法的特异性为100%,灵敏度检测线为1%。结论本研究成功建立牛源性肉制品的检测方法,该方法快速简便,且具有较高的特异性和灵敏度,可用于市售肉制品中牛源性成分的鉴定。     

实时荧光PCR法检测肉制品中猪源性成分

目的S建立一个高特异性和高灵敏度的有效检测猪源性成分的检测方法。方法S以猪雌激素受体基因设计合成一对特异性引物,采用液氮研磨结合试剂盒抽提的方法提取猪肉中的DNA,分别以300S,30S,3S,0.3S和0.03Sng为模板量进行梯度反应,基于荧光定量PCR技术(SYBR法)建立检测方法。结果S以Ct值作为纵坐标,以lgXS(X为DNA的量,ng)的值作为横坐标,得到标准曲线方法Ct=30.473-3.542lgX,SR2=0.9997;该方法的检测灵敏度达到2 Spg。结论S该猪源性成分检测方法简单快捷,特异性和重复性好,灵敏度高,可作为市场监督和检测鉴定的可行性方法。     

利用实时荧光定量PCR法检测食品中鹌鹑源性成分

为建立基于TaqMan实时荧光聚合酶链式反应(polymerase chain reaction,PCR)技术在食品中的鹌鹑源性成分的定量检测方法,根据现行检验检疫标准(SN/T 2727-2010)合成引物及探针。首先对13种不同动物鲜肉组织的DNA进行鹌鹑源性成分特异性检测,然后对鹌鹑源性DNA模板原液进行梯度稀释,检测方法灵敏度,最后在加工制品中检测方法的适用性。研究结果表明:建立的方法特异性强,除鹌鹑肉外,牛、羊、猪、马、驴、狗、兔子、鸡、鸭、鸽子、火鸡、鱼12种动物鲜肉组织均无特异性扩增;方法的灵敏度较高,鹌鹑组分DNA的检出限可达10 fg/mL,灵敏度可达0.001%;方法的适用性较广,可以用于加工制品中鹌鹑源性成分的检测。     

双重PCR检测食品中的动物源性成分

建立了一种检测肉制品中动物源性成分的双重PCR技术,针对五种不同动物线粒体DNA中所含有的特异性基因分别设计引物,同时对脊椎动物所共有的保守基因进行引物设计,并将其作为体系中的内参照。用此技术可同时检测出肉制品中5种动物源性成分,检测灵敏度达到1%,能够适应市场化检测的需要。      

双重PCR检测食品中的动物源性成分

建立了一种检测肉制品中动物源性成分的双重PCR技术,针对五种不同动物线粒体DNA中所含有的特异性基因分别设计引物,同时对脊椎动物所共有的保守基因进行引物设计,并将其作为体系中的内参照.用此技术可同时检测出肉制品中5种动物源性成分,检测灵敏度达到1%,能够适应市场化检测的需要.     

实时荧光PCR法鉴别莜面中掺杂淀粉的植物源性

以红薯ACtin基因、马铃薯UGPase基因、玉米ZEIN基因、木薯g3pdh基因设计种属特异性引物探针,以莜麦Avenin基因、真核生物18S rRNA为对照,建立一种实时荧光PCR法对莜面掺杂植物源性淀粉进行鉴别。该方法特异性强,相对灵敏度为0.1%,可用于莜面中掺杂植物源性淀粉的鉴别。     

动物源性食品PCR检测方法的应用研究

动物源性食品的成分检测已成为当前食品质量检测工作的一个重要环节,PCR检测方法在该检测中得到了较为广泛的应用。基于此,本文对PCR在动物源性食品检测中的应用进行了探讨,通过对市售多种肉制品进行PCR检测实验,对PCR检测方法的流程和要点进行详细阐述,以期为今后的相关检测工作提供参考。     

肉制品中鸭源性成分的实时荧光PCR检测

目的：建立基于实时荧光PCR技术的鸭源性成分的快速检测方法。方法：以鸭线粒体DNA序列为目的基因,设计并筛选了鸭源性特异性引物及探针,进行荧光定量PCR扩增,建立鸭源性成分检测方法。通过特异性、灵敏性、及盲样检测试验,对该体系进行验证。结果：该方法能够快速有效的检测鸭源性成分,具有较强的特异性及灵敏性,灵敏度约为0．01％（质量分数）;通过市售盲样肉制品的检测,表明该体系可用于定性检测加工肉制品中的鸭源性成分。结论：该方法特异性强,灵敏度高,可用于对肉制品中鸭源性成分的掺假鉴别。     

PCR法检测肉制品中鹅源性成分

目的 建立一种PCR法检测肉制品中鹅源性成分的方法。方法 根据鹅的线粒体DNA(mtDNA)序列设计引物, 以9种动物的DNA为模板, 利用新设计的鹅的特异性引物进行PCR反应, 并用琼脂糖凝胶电泳检测引物的特异性和敏感性。结果 通过PCR反应, 仅鹅的DNA扩增得到了194 bp的目的片段, 其余物种DNA和空白对照均无目的片段。扩增产物的核苷酸序列与GENBANK中检索到的相应序列基本相符合。 结论 该方法特异性强、灵敏度高, 适合检测肉制品中的鹅源性成分。     

A comparison of the flavour volatiles from cooked beef and pork meat systems

The headspace volatiles from beef and pork cooked with and without added adipose tissue were entrained on Tenax GC before analysis by gas chromatography (g.c.) and combined gas chromatography-mass spectrometry. Lipid-derived volatiles dominated the headspaces of all samples and the only qualitative difference between beef and pork was an alkene of lipid origin which was peculiar to beef samples. A comparison of the areas of 37 g.c. peaks from replicate analyses of the different meat preparations using canonical variates and stepwise discriminant statistical techniques unambiguously distinguished the lean meats and the lean meats cooked with adipose tissue of either species. Adding adipose tissue to lean meat did not result in proportional increases in lipid-derived volatiles.     

The tenderness of cooked and raw meat from young and old beef animals

The shortening-toughness relationships for sternomandibularis muscles of young ox and large old bull have been compared. In both, toughness, measured by tenderometer, increases to a maximum at a shortening of 40%, and declines steeply from there at higher shortenings. This characteristic peaked relationship is obtained for ox and bull cooked both at 60 and 80 °C. However, toughness values for meat cooked to 60 °C are only about half those for meat cooked to 80 °C. In contrast to these similarities in the relationships for young and old animals, the rate of toughness increase with shortening in ox is considerably less than in bull. Thus toughness of ox at a given shortening and cooking temperature is considerably lower than that for bull, especially at high shortenings. A further distinction is made between the animals in that raw ox muscle does not give a peaked shortening-toughness relationship, whereas raw old bull does. The results have shown that for longissimus and sternomandibularis muscles a trained taste panel is only sensitive to variations in toughness in the lower half of the range of shear force values determined by tenderometer. In the light of this, live animal characteristics and muscle shortening are both important in determining toughness measured by taste panel.     

Comparison of instrumental and sensory evaluation of texture of cured and cooked beef meat

The instrumental and sensory texture attributes of beef muscles (M. longissimus dorsi and M. semimembranosus) were compared after curing and thermal treatments. Shear, compression and puncture tests were carried out with an UTM Instron 4301 and the sensory evaluation was made with the score method. The force values obtained for puncture test gave a greater degree of correlation with the sensory tenderness, hardness and elasticity than the shear test forces. The shear test forces were found to significantly correlate with sensory tenderness only for muscles with perpendicular orientation of fibres to the direction of shear blade movement. The evaluation of beef texture with compression test was dependent on the level of sample deformation degree. The force values of compression were found to correlate significantly with elasticity and juiciness of meat up to 60% deformation levels, but at deformation levels higher than 60% appeared significant correlation with tenderness and hardness. The obtained results showed the usefulness of puncture test with proposed parameters for the instrumental measurements of beef texture.     

Detection of chicken and turkey meat in meat mixtures by using real-time PCR assays

In this study, TaqMan-based real-time Polymerase Chain Reaction (PCR) techniques were developed for the detection of chicken and turkey meat in raw and heat-treated meat mixtures. Primers and TaqMan probe sets were designed to amplify 86 bp and 136 bp fragments for the chicken and turkey species, respectively, on the mitochondrial NADH dehydrogenase subunit 2 gene. In the results, it was possible to detect each species at the level of 0.1 pg template DNA with the TaqMan probe technique without any cross-reactivity with nontarget species (bovine, ovine, donkey, pork, and horse) while the detection level was 1 pg template DNA using conventional PCR. The TaqMan probe assays used in this study allowed the detection of as little as 0.001% level of both species in the experimental meat mixtures, prepared by mixing chicken and turkey meat with beef at different levels (0.001% to 10%). In conclusion, TaqMan probe assays developed in this research are promising tools in the specific identification and sensitive quantification of meat species even in the case of heat-treated meat products, and suitable for a rapid, automated, and routine analysis.     

多重PCR法用于畜肉源性鉴定的研究

根据Genebank中猪、牛、羊的线粒体细胞色素b（Cyt-b）基因序列,设计了猪、牛、羊的通用上游引物和特异性下游引物,通过调节引物比例、反应体系、反应条件以及模板量等优化实验确立了多重PCR检测方法。研究结果显示,该多重PCR方法能够同时扩增样品中猪、牛、羊成分,对混合样品的检出限可达10%;并将该方法用于成都市猪、牛、羊肉的调查研究,得到了理想的结果。      

多重PCR法用于畜肉源性鉴定的研究

根据Genebank中猪、牛、羊的线粒体细胞色素b(Cyt-b)基因序列,设计了猪、牛、羊的通用上游引物和特异性下游引物,通过调节引物比例、反应体系、反应条件以及模板量等优化实验确立了多重PCR检测方法.研究结果显示,该多重PCR方法能够同时扩增样品中猪、牛、羊成分,对混合样品的检出限可达10％;并将该方法用于成都市猪、牛、羊肉的调查研究,得到了理想的结果.     

微滴数字PCR方法检测畜肉食品中鸭源性成分

目的利用微滴数字PCR技术,建立鸭源性成分的快速、特异、灵敏检测方法。方法根据鸭线粒体基因全序列设计的引物及探针,利用微滴数字PCR进行扩增,建立鸭源性成分检测方法;以市场中常见畜禽肉为参考物种作微滴数字PCR特异性检测;以羊肉为干扰物观察微滴数字PCR的抗干扰性;将鸭肉DNA进行梯度稀释,对方法灵敏度进行检测。结果该方法能够有效对鸭源性成分进行检测,特异性强,灵敏度高,并且其他物种成分的存在对方法灵敏度检测没有影响。结论该方法特异性强,灵敏度高,可以实现畜肉食品中的鸭源性成分检测。