基于TaqMan实时荧光PCR检测鲜肉及加工肉制品中的驴源性成分

根据出入境检验检疫行业标准SN/T 3730.4—2013《食品及饲料中常见畜类品种的鉴定方法 第4部分：驴 成分检测 实时荧光PCR法》合成引物和探针,利用TaqMan实时荧光聚合酶链式反应（polymerase chain reaction, PCR）技术检测鲜肉及加工肉制品中的驴源性成分。首先对13 种不同动物鲜肉组织的DNA进行驴源性成分特异 性检测,然后对驴源性DNA模板原液进行梯度稀释,检测方法灵敏度,最后在加工肉制品中检测方法的适用性。 结果表明：本研究建立的方法特异性强,除驴肉外,牛、羊、猪、马、骆驼、鹿、狗、兔、鸡、鸭、鸽子、鹌鹑 12 种动物鲜肉组织均无特异性扩增;方法的灵敏度较高,驴组分DNA的检出限可达100 fg/μL,灵敏度可达0.01%; 方法的适用性较广,可以用于加工肉制品中驴源性成分的检测。     

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。     

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。      

食品中鸡源性成分标准检测方法的比较性研究

为比较食品中鸡源性成分的标准检测方法,本试验对3种标准检验的引物计、方法及灵敏度进行比较.结果 显示,实时荧光PCR法引物特异性好、方法简便、灵敏度高.因此得出结论,两种实时荧光PCR法均可作为鸡源性成分检测的补充方法.     

GNM C7-8实时荧光PCR系统对掺假肉制品中多种动物源性成分的同时检测

应用GNM C7-8实时荧光PCR系统,同时快速检测掺假肉制品中多种动物源性成分。将牛肉粉、驴肉粉、鸵鸟肉粉和羊肉粉4组不同肉粉中分别混入猪肉粉、鸭肉粉、马肉粉、鸡肉粉和狐狸肉粉中的2~3种肉粉,制备人工模拟掺假肉制品。通过GNM C7-8实时荧光PCR和ABI7500荧光PCR方法,对上述四种掺假肉制品进行多种动物源性成分检测和比较。结果表明,GNM C7-8实时荧光PCR方法与ABI7500荧光PCR方法从掺假肉制品中检测猪、鸭、马、鸡和狐狸5种成分的循环数一致,分别是31、31、29、32和25,但基于八模块设计的GNM C7-8实时荧光PCR方法能同时检测四种掺假肉制品中不同动物源性成分,检测时间更短。因此,GNM C7-8实时荧光PCR系统的八个模块能够独立运行,互不干扰,无交叉污染,能够实现对掺假肉制品中多种动物源性成分的同时快速检测,为肉制品掺假检测提供强有力的方法支持。     

素食中动物源性成分的PCR检测方法的研究

建立了素食产品中动物源性成分的实时荧光PCR检测方法,该方法特异、灵敏,仅通过检测一个基因位点即可判断素食产品中是否含有动物源性成分,对来源于畜类、禽类及鱼类动物源性成分的检出限分别为0.01%,0.05%,0.1%。运用该方法对市场上22份不同类型的素食产品进行了检测,检出2份产品中含有动物源性成分。该方法操作方便快捷,适用于素食中动物源性成分的定性检测。     

肉及肉制品中动物源性成分核酸检测方法研究进展

民以食为天,食以安为先。肉类为人类的生命活动提供了丰富的蛋白质、脂肪和B族维生素等,是人们生活的重要食品。但在利益的驱动下肉制品行业内掺假事件时有发生,严重侵害了消费者权益,并造成了不良的经济和社会影响。因此加强研究肉类产品动物源性成分的检测技术,广泛开展肉及肉制品中动物源性成分的检验意义重大。本文综合论述了目前已有的肉及肉制品中动物源性成分的核酸检测技术,并对有应用前景的新技术做了简要介绍。     

GNM C7-8实时荧光PCR同时快速检测8种动物源性成分

目的基于八模块设计的GNMC7-8实时荧光PCR系统实现对8种动物源性成分同时快速检测。方法基于猪朊蛋白基因、牛生长激素基因、羊生长激素基因、驴线粒体ATPase 6基因、马线粒体ATPase 6基因、鸭生长激素基因、水貂线粒体细胞色素b基因、狐狸线粒体细胞色素氧化酶Ⅰ亚基基因引物设计了8种动物源性成分实时荧光PCR检测方法。应用GNMC7-8实时荧光PCR系统,对8种动物源性成分启动同时检测和每隔5min顺序启动不同时检测,并将检测结果和ABI7500荧光PCR分别检测结果进行比较。结果 GNM C7-8实时荧光PCR系统同时检测猪、牛、羊、驴、马、鸭、水貂、狐狸8种动物源性成分,起峰周期数分别为30、27、21、21、24、17、15、17,与其不同时检测结果一样,与ABI7500荧光PCR系统检测结果相比,猪、羊、马、鸭、水貂5种成分的起峰快1～2个周期数,其余3种成分检测结果一致,检测完成时间少16～20 min。结论 GNM C7-8实时荧光PCR系统8个模块独立运行,互不干扰,无交叉污染,能够同时快速检测8种动物源性成分,为快速筛查动物源性成分提供强有力的设备技术支持。     

肉制品中动物源性成分DNA检测方法的研究进展

肉制品主要成分标识的真实性是全球重要的食品安全问题之一,特别是肉制品中动物源性成分的掺假和标识问题已引发全球关注。如何对肉制品中动物源性成分进行鉴定和标识已成为产品真实性鉴定的热点。基于DNA分子稳定性强的优点,DNA检测技术被广泛用于食品安全检测和监测诸多领域,体现出了灵敏度高、特异性强等优势。本文重点从动物源性检测的靶序列DNA选择和DNA分析技术研究2个方面,阐述了肉制品中动物源性成分定性、定量检测技术的研究和应用,并讨论动物源性成分定量分析的可能性,为我国实施动物源性成分量化监管提供思路。     

环介导等温扩增与qPCR用于检测肉制品中狗源性成分的比较

目的 建立基于实时荧光PCR和环介导等温扩增检测肉制品中狗源性成分的检测方法并比较2种方法的检测效能。方法 针对狗线粒体CYTB基因保守序列, 采用Primer Explorer Version 4软件设计环介导等温扩增引物, Primer Express 3.0.1软件设计实时荧光PCR引物及探针。提取狗肉基因组DNA作为阳性模板, 黄牛肉等20种基因组DNA作为阴性对照, 分析扩增特异性; 用含靶序列的人工质粒确定检测灵敏度及人工掺肉样确定最低检出限; 用市售肉制品分析检出率。结果 对于肉制品中狗源性成分检测, 实时荧光PCR和环介导等温扩增检测均有较好的特异性, 人工质粒分析灵敏度分别为0.1、1 fg/μL。对10种市售狗肉制品的狗源性成分均检出阳性、10份不含狗肉制品均检出阴性, qPCR法和环介导等温扩增法的检测结果符合率均为100%。结论 qPCR法和环介导等温扩增法在检测肉及肉制品中的狗源性成分时, 在灵敏度、检出限和准确度上具有相当的效能。     

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。     

UPLC-ESI-MS/MS analysis of Sudan dyes and Para Red in food

An analytical method for the simultaneous determination of Sudan dyes (Sudan Red G, Sudan I, Sudan II, Sudan III, Sudan Red 7B and Sudan IV) and Para Red in food by ultra-performance liquid chromatography-electrospray tandem mass spectrometry (UPLC-ESI-MS/MS) was developed. Samples were extracted with acetonitrile, and water added into the extract. The supernatant was analysed by UPLC-MS/MS after refrigeration and centrifugation. The sample was separated on an Acquity BEH C₁₈ column, and detected by MS/MS with the multiple reaction monitoring mode. Matrix calibration was used for quantitative testing of the method. The linear matrix calibrations of Sudan dyes and Para Red were 2–50 and 10–250 ng g^?1, respectively, and the regression coefficients were >0.9945. The recoveries were 83.4–112.3% with good coefficients of variation of 2.0–10.8%. The limits of detection were between 0.3 and 1.4 ng g^?1 for the six Sudan dyes, and between 3.7 and 6.0 ng g^?1 for Para Red. The limits of quantification were between 0.9 and 4.8 ng g^?1 for the six Sudan dyes, and between 12.2 and 19.8 ng g^?1 for Para Red.     

Dietary exposure of secondary school students in Hong Kong to polybrominated diphenyl ethers from foods of animal origin

This paper reports levels of polybrominated diphenyl ethers (PBDEs) in a wide range of foods of animal origin and estimates their dietary exposure for secondary school students in Hong Kong, China. Dietary exposure to PBDEs was estimated using local food consumption data obtained from secondary school students in 2000 and the concentrations of PBDEs in food samples taken from local market in 2008. The PBDE levels on a fresh weight basis for fish ranged from 13 to 6600 pg g^?1, for seafood and seafood products ranged from 15 to 1200 pg g^?1, for meat and meat products ranged from 23 to 3500 pg g^?1, for poultry ranged from 68 to 670 pg g^?1, for eggs ranged from 280 to 800 pg g^?1, and for dairy products ranged from 12 to 480 pg g^?1. The dietary exposures of secondary school students for the average and high consumers were estimated to be 2.6 and 6.4 ng kg^?1 body weight day^?1, respectively. According to the Joint FAO/WHO Expert Committee on Food Additives (JECFA), for the more toxic PBDE congeners, adverse effects would be unlikely to occur in laboratory animals at doses of less than approximately 100 µg kg^?1 body weight day^?1. The resulting margins of exposures (38,000 for average consumers and 16,000 for high consumers) showed that the estimated dietary exposures of secondary school students were far below any adverse effect dose observed in laboratory animals and were therefore of low concern for human health.     

2011—2016年广西壮族自治区市售肉及肉制品食源性致病菌污染状况分析

目的 了解广西壮族自治区市售肉及肉制品中食源性致病菌污染状况,为有效开展食源性疾病防控提供科学依据。方法 2011—2016年从广西壮族自治区14个市采集5类市售肉及肉制品共10 927份,按照国家标准方法进行9种食源性致病菌检验。结果 10 927份样品的食源性致病菌总检出率为5.0%(548/10 927),食源性致病菌检出率按样品种类依次为调理肉制品(33.3%,33/99)、生畜肉(24.5%,73/298)、生禽肉(24.2%,67/277)、冷冻肉糜制品(14.4%,14/97)、熟肉制品(3.6%,361/10 156)。检出的主要致病菌为沙门菌、金黄色葡萄球菌和单核细胞增生李斯特菌。生禽肉中未检出弯曲菌和致泻大肠埃希菌,调理肉制品中未检出致泻大肠埃希菌,熟肉制品中未检出志贺菌。熟肉制品各年度检出率范围为0.9%～4.9%。结论 广西壮族自治区市售肉及肉制品受到不同程度食源性致病菌污染,且污染持续多年存在。     

Multi-residue method for the confirmation of four avermectin residues in food products of animal origin by ultra-performance liquid chromatography–tandem mass spectrometry

A confirmatory method was developed for the rapid determination of abamectin, ivermectin, doramectin and eprinomectin residues in various food products of animal origin, such as pork muscle, pork liver, fish and milk. Samples were homogenized, extracted and de-proteinized by acetonitrile, cleaned via two-step cleaning procedure using Bond Elut C₁₈ SPE columns and then alumina-N cartridges. All the four avermectin residues in different animal-food products were simultaneously separated and determined by ultra-performance liquid chromatography–electrospray ionization tandem mass spectrometry (UPLC–ESI–MS/MS) within 3.5?min. Data acquisition under positive ESI–MS/MS was performed by applying multiple reaction monitoring (MRM) for both identification and quantification, and mass spectrometric conditions were optimized to increase selectivity and sensitivity. The matrix-matched calibration curves for different matrices, such as pork muscle, pork liver, fish and milk, were constructed and the interference effect of different sample matrices on the ionization was effectively eliminated. The UPLC–MS/MS method was validated with satisfactory linearity, recovery, precision and stability. Matrix-matched calibration curves of abamectin, ivermectin, doramectin and eprinomectin in four different matrices were linear (r^{2 ?}≥?0.990, goodness-of-fit coefficients ≤12.8%) in the range 2.5–200?µg?kg^?1. The limits of detection and quantification for the four avermectins were in the range 0.05–0.68 and 0.17–2.27?µg?kg^?1, respectively. Recoveries were 62.4–104.5% with good intra- and inter-day precision. The method was rapid, sensitive and reliable, and can be applied to the quantitative analysis of avermectin residues in different animal-food products.     

Screening for tetracycline residues in food products of animal origin in the State of Kuwait using Charm II radio-immunoassay and LC/MS/MS methods

This study surveyed tetracycline residues in foods marketed in Kuwait using various techniques to determine their prevalence above the threshold level. A total of 1517 locally produced and imported food samples of animal origin were collected for analyses, comprising dairy products, eggs and tissue samples (meat, poultry and fish) to give a representative picture of the current use and/or misuse of the drug in Kuwait. Screening was carried out using the Charm II test for tetracycline residues. Positive and suspect positive samples were confirmed by LC/MS/MS. Negative and positive controls, in triplicate, were applied to each method and showed 80–100% agreement. The results showed that 100% of tested eggs, meat, fish, ice cream and cheese were within the limit, while 5% of poultry and 18% of milk samples were above the permitted limit.