基于肽生物标志物技术鉴别肉品的真实性研究进展

基于DNA和蛋白质的方法已广泛应用于肉制品的真实性鉴定,但肉制品经加工处理后,DNA和蛋白质很容易发生降解和变性,而蛋白质的一级结构(氨基酸序列)则足够稳定。伴随着蛋白质组学的发展,以肽生物标志物为基础的鉴定方法逐渐应用于肉制品的真实性分析。本文将从肉中肽生物标志物的分离鉴定方法、肽生物标志物在鉴定不同肉制品及肉制品中外源蛋白的应用三个方面进行归纳和总结,并对肽生物标志物在鉴定肉品真实性应用中的发展方向进行展望,以期为食品溯源提供更丰富的技术手段。     

Multiplex PCR in Species Authentication: Probability and Prospects—A Review

Food forgery is one of the most articulated socio-economic concerns which contributed to increase people’s awareness on what they eat and how and where it is produced. Consumers are anxious about the consequences of food falsification on their choices, religious rituals, health, and hard-earned fortunes. The recent scandals of horse and rat meats in Europe and China have given us a brainstorming apprehension on the detection, differentiation, and identification of meat products. To restore consumers’ trust and protect wildlife in natural habitats, researchers and policy-making and policy-implementing authorities have massively monitored all steps in the production of foods and food materials. Analytical approaches based on lipids, proteins, and DNA have been proposed for the authentication of meat species under pure and complex matrices. However, protein and lipid-based methods are less effective since the target biomarkers could be modified throughout the processing treatments. On the other hand, DNA-based species identification schemes have gained wider acceptance and reliability because of the superior stability and universality of DNA in all tissues and cells. We systematically presented here major species detection schemes with special emphasis on multiplex polymerase chain reaction (PCR) of both end-point and real-time platforms. We believe this short but comprehensive review would serve as a reference guide for the developers and users of multiplex PCR and others DNA-based techniques.     

畜禽肉中源成分检测技术研究进展

近年来,畜禽肉掺假问题严重侵害消费者权益,引发公众对食品安全的担忧及市场监管部门的关注.禽畜肉溯源分析和鉴别技术成为食品领域研究热点,畜禽肉中的源成分是用于区分鉴定不同物种的定性成分,在肉制品质量检测及肉制品真实性鉴别方面应用广泛.本文针对目前定性及定量检测禽畜肉中源成分的检测技术,主要包括基于蛋白质的检测技术、基于脂...     

Analytical approaches of meat authentication in food

Meat industry sector is likely exposed to illegal practices which involve replacement of meat ingredients with other animal species. Meat products are commonly modified during processing, and the cases have been rampant in certain part of the world. Meanwhile, individual consumption patterns are affected by religious belief and health reason. For instance, Muslims and Jews are prohibited from consuming pork and allergic reaction to certain meat species affects choices of meat products. Thus, food authentication is vital as it offers protection to consumers from fraudulent act. Numerous authentication techniques have been developed to tackle food fraud issues. Authentication methods are categorised mainly as targeted analysis which provides quantitative outcome and non-targeted analysis which offers qualitative result. In the present review, both targeted and non-targeted analyses for meat species authentication are discussed based on previous research conducted using these methods and how they serve the purpose for authentication.     

DNA条形码技术在食品鉴定中的应用

近年来,国内外不断出现有关食品掺假造假等食品安全问题的报道。产品与标签不符,肉类掺假,以次充好等商业欺诈问题影响着人们的利益与健康。食品掺假造假问题越来越受到重视,而传统的检测方法已不能满足食品鉴定的需要。DNA条形码技术是从分子水平上对食品进行鉴定,弥补了传统鉴定方法的不足,其准确、高效、简单的特点为食品鉴定领域带来了新的革命。本文在简要介绍DNA条形码研究现状的基础上,主要总结目前国内外关于DNA条形码技术在食品鉴定中的应用情况,包括在渔类产品、肉类产品、可食用植物、加工食品鉴定中的应用,最后讨论DNA条形码技术在食品鉴定中的优缺点以及发展趋势。     

微滴式数字聚合酶链式反应精准定量检测羊肉中掺杂猪肉

以羊和猪的单拷贝持家基因DNA复制蛋白A1为靶基因设计合成了适用于微滴式数字聚合酶链式反应的特异性引物和探针,通过理论推导获得了单位质量两种肉基因拷贝数之比的固定值,并进行了验证,据此将样品中羊肉和猪肉的拷贝数转换为相对质量分数,从而建立了羊肉中掺杂猪肉的精准定量检测方法。该方法可以很好地应用于羊肉中掺杂猪肉的含量检测,猪肉的最低定量限为1%,在5%~80%范围内绝对误差小于±1.30%,相对误差小于±10%,定量结果准确、重复性高,可以为肉类掺假的监管工作提供有力的技术参考。     

Authentication technologies using DNA-based approaches for meats and halal meats determination

Food authenticity and safety are major concerns for researchers, consumers, and particularly the meat industry. Meat products are targets for species substitution and adulteration due to their market value. Presently, the demand for halal products is witnessing a substantial increase. Therefore, it is essential to use appropriate science-based methods for determining the species origin of halal meat. DNA barcoding is a useful technique for the molecular identification of biological specimens, and raw and processed foods. The potential of using DNA barcoding is increasingly applied as an authentication tool for halal animal and meat products. Our review will bring together all DNA-based techniques that have been developed for the authenticity of meat derived from halal and non-halal animals and also their derivatives. Additionally, the present paper will highlight the possibility of using the DNA barcoding approach for halal meat authenticity.     

A fast and accurate method for controlling the correct labeling of products containing buffalo meat using High Resolution Melting (HRM) analysis

The substitution of high priced meat with low cost ones and the fraudulent labeling of meat products make the identification and traceability of meat species and their processed products in the food chain important. A polymerase chain reaction followed by a High Resolution Melting (HRM) analysis was developed for species specific detection of buffalo; it was applied in six commercial meat products. A pair of specific 12S and universal 18S rRNA primers were employed and yielded DNA fragments of 220 bp and 77 bp, respectively. All tested products were found to contain buffalo meat and presented melting curves with at least two visible inflection points derived from the amplicons of the 12S specific and 18S universal primers. The presence of buffalo meat in meat products and the adulteration of buffalo products with unknown species were established down to a level of 0.1%. HRM was proven to be a fast and accurate technique for authentication testing of meat products.     

Peptide biomarkers as a way to determine meat authenticity

Meat fraud implies many illegal procedures affecting the composition of meat and meat products, something that is commonly done with the aim to increase profit. These practices need to be controlled by legal authorities by means of robust, accurate and sensitive methodologies capable to assure that fraudulent or accidental mislabelling does not arise. Common strategies traditionally used to assess meat authenticity have been based on methods such as chemometric analysis of a large set of data analysis, immunoassays or DNA analysis. The identification of peptide biomarkers specific of a particular meat species, tissue or ingredient by proteomic technologies constitutes an interesting and promising alternative to existing methodologies due to its high discriminating power, robustness and sensitivity. The possibility to develop standardized protein extraction protocols, together with the considerably higher resistance of peptide sequences to food processing as compared to DNA sequences, would overcome some of the limitations currently existing for quantitative determinations of highly processed food samples. The use of routine mass spectrometry equipment would make the technology suitable for control laboratories.     

一对同时鉴定8种动物源性成分的通用引物的制备及应用

肉类真假鉴定是食品检测工作的内容之一,目前已有多种基于PCR的肉类鉴定方法,但是鉴定种类和效率受限。本研究设计了一对基于普通PCR技术可同时鉴定8种动物源性成分的通用引物并建立了鉴定方法。该引物以线粒体DNA为靶标,利用扩增产物中不同物种间的插入缺失多态性片段大小即可鉴定山羊、绵羊、鹿、水牛、牛、牦牛、猪和骆驼8个物种,扩增后分别得到728 bp、704 bp、504 bp、453 bp、448 bp、431 bp、396 bp和326 bp的片段,每种PCR产物经SspI酶切后产生数量和大小不同的片段,可以进一步清晰鉴别8个物种。引物特异性测试表明和其他常见肉类动物DNA无交叉反应,DNA检测最低限度在0.01~0.05 ng。应用本方法对40份市场肉类及产品的检测表明,羊肉串、羊肉卷以及特色畜产品如驼肉、鹿肉和驴肉存在较多的掺假行为。与其他现有PCR检测方法相比,该方法具有简便易行和高通量的优点,可以作为肉类掺假筛选检测的常规方法。     

Potential DNA markers as a rapid tracing tool for animal adulterants in vegetarian food

DNA-based methods are rapid, cost-effective and broadly applicable approaches for food authentication. Recently, the requirements for food safety and food integrity have increased with improved quality of life. Methodologies regarding food authentication based on DNA analysis are more commonly being used. With the increasing number of vegetarians, searching for markers for blind identification across kingdom species, such as an ingredient of animal origin in vegetarian food, would be valuable and attractive. Using bioinformatic analysis of an existing data source composed of 481 ultraconserved sequences, we selected 6 new candidate DNA segments that exist in most vertebrates but that do not exist in plants. Then, primers were designed for all of the candidate DNA markers, and DNA samples isolated from cow, pig, chicken, duck, soy bean, rice, pepper, wheat, sunflower and colza were amplified using each primer pair. None of the plant DNA samples generated a PCR product, while the DNA samples of animal origin were amplified successfully using 5 of the candidate segment primers; the 6th segment primer failed to amplify the DNA and was discarded. Moreover, a simulation experiment containing a plant product contaminated by an animal component indicated that the candidate DNA markers can be used for the rapid detection of animal adulterants in vegetarian products with a promising 5% detection limit. The identified candidate DNA markers for the blind identification of animal adulteration in vegetarian food may be highly desirable in the vegetarian food market, and these markers may facilitate the study of molecular technology for food authentication.     

Species determination – Can we detect and quantify meat adulteration?

Proper labelling of meat products is important to help fair-trade, and to enable consumers to make informed choices. However, it has been shown that labelling of species, expressed as weight/weight (w/w), on meat product labels was incorrect in more than 20% of cases. Enforcement of labelling regulations requires reliable analytical methods. Analytical methods are often based on protein or DNA measurements, which are not directly comparable to labelled meat expressed as w/w. This review discusses a wide range of analytical methods with focus on their ability to quantify and their limits of detection (LOD). In particular, problems associated with a correlation from quantitative DNA based results to meat content (w/w) are discussed. The hope is to make researchers aware of the problems of expressing DNA results as meat content (w/w) in order to find better alternatives. One alternative is to express DNA results as genome/genome equivalents.     

Detection of allergenic additives in processed meat products

Allergic responses to food components are an increasing problem all over the world. It is therefore important to protect people who are vulnerable to food allergens against accidental and unintended consumption of products containing allergic ingredients. The meat industry commonly uses various allergic additives in the production of processed products, such as legumes (soy, peas, beans), milk and egg preparations, cereals containing gluten (wheat, rye, barley and oats), and spices (celery and mustard). These meat additives have specific technological properties, which help to create a texture or flavor profile, or affect the nutritional value, although some of them, such as soy, mustard, milk and egg white proteins, can cause severe allergic reactions. The aim of this paper is to discuss the application of various recently established methods of detection of allergenic additives in processed meat products – for instance cold cuts and sausages. The new methods are based mainly on protein, DNA, and isoflavones or phytic acid analysis. The article also characterizes the latest trends in the development of research on methods that would enable quick and reliable identification of targeted allergens in meat products. © 2018 Society of Chemical Industry     

Authenticity Determination of Meat and Meat Products on the Protein and DNA Basis

Adulterated food can be defined as food incompatible with the declaration of the seller. In the case of meat and meat articles, adulterations refer not only to the replacement of ingredients but also to inappropriate information concerning the origin of raw materials. Methods aiming at investigating meat and meat product authenticity may be based either on the analysis of protein composition or on the analysis of nucleic acids. At the present time, meat and meat product authenticity investigations based on protein analysis employ electrophoretic, enzymic, and chromatographic methods, sometimes supported by the mass spectrometry technique. On the other hand, species identification is often based on polymerase chain reaction (PCR). Biochips present a promising technology.     

Methodology for meat species identification: A review

This review covers methods available for the recognition of the species of animal whose meat is present in raw materials or meat products. Isolation of species-specific proteins is cumbersome; however, isoelettric focusing followed by recognition of the pattern of protein bands is particularly effective for speciation. Immunoassay has been established as a powerful technique for the determination of food protein analytes in suitable extracts of unresolved mixtures. Simple immunodiffusion using antisera to serum proteins is sufficient to provide a check on the identity of suspect raw samples; routine control would involve extensive sampling programmes. Enzyme-linked immunosorbent assay procedures are also appropriate, but attempts to quantify individual meat species in mixtures have not been successful due to the variability of residual blood levels. Species identification in heat-processed meat products is hindered by progressive denaturation of the protein markers, leading to loss of solubility and antigenicity; the effects of heating can be minimised by a judicious choice of analyte and its solubilisation by renaturation.     

Universal primers for species authentication of animal foodstuff in a single polymerase chain reaction

BACKGROUND: There are many DNA‐based systems for detecting animal species present in food and food products, applicable for food quality control and authentication. However, most (if not all) methods require more than one pair of primers and cannot be applied over a wide taxonomic range, e.g. identifying vertebrates and invertebrates with the same primers and protocols. RESULTS: A pair of primers is described here that allows in a single polymerase chain reaction the identification of animal species in food and processed (precooked, canned or smoked) food products over a wide taxonomic range. CONCLUSION: These primers permit the identification of most animal taxa employed in human nutrition, from invertebrates such as molluscs to higher vertebrates, distinguishing between species of the same genus. The short fragment amplified within the 16S rDNA exhibits phylogenetic value and could be considered universal based on the wide taxonomic range assayed. The primers are easy to use and accessible for laboratories with a modest budget, as well as being valuable for consumer information and to reveal food fraud. Copyright © 2012 Society of Chemical Industry     

Authentication of berries and berry-based food products

Berries represent one of the most important and high-valued group of modern-day health-beneficial “superfoods” whose dietary consumption has been recognized to be beneficial for human health for a long time. In addition to being delicious, berries are rich in nutrients, vitamins, and several bioactive compounds, including carotenoids, flavonoids, phenolic acids, and hydrolysable tannins. However, due to their high value, berries and berry-based products are often subject to fraudulent adulteration, commonly for economical gain, but also unintentionally due to misidentification of species. Deliberate adulteration often comprises the substitution of high-value berries with lower value counterparts and mislabeling of product contents. As adulteration is deceptive toward customers and presents a risk for public health, food authentication through different methods is applied as a countermeasure. Although many authentication methods have been developed in terms of fast, sensitive, reliable, and low-cost analysis and have been applied in the authentication of a myriad of food products and species, their application on berries and berry-based products is still limited. The present review provides an overview of the development and application of analytical chemistry methods, such as isotope ratio analysis, liquid and gas chromatography, spectroscopy, as well as DNA-based methods and electronic sensors, for the authentication of berries and berry-based food products. We provide an overview of the earlier use and recent advances of these methods, as well as discuss the advances and drawbacks related to their application.     

Gelatin controversies in food,pharmaceuticals, and personal care products: Authentication methods,current status,and future challenges

Gelatin is a highly purified animal protein of pig, cow, and fish origins and is extensively used in food, pharmaceuticals, and personal care products. However, the acceptability of gelatin products greatly depends on the animal sources of the gelatin. Porcine and bovine gelatins have attractive features but limited acceptance because of religious prohibitions and potential zoonotic threats, whereas fish gelatin is welcomed in all religions and cultures. Thus, source authentication is a must for gelatin products but it is greatly challenging due to the breakdown of both protein and DNA biomarkers in processed gelatins. Therefore, several methods have been proposed for gelatin identification, but a comprehensive and systematic document that includes all of the techniques does not exist. This up-to-date review addresses this research gap and presents, in an accessible format, the major gelatin source authentication techniques, which are primarily nucleic acid and protein based. Instead of presenting these methods in paragraph form which needs much attention in reading, the major methods are schematically depicted, and their comparative features are tabulated. Future technologies are forecasted, and challenges are outlined. Overall, this review paper has the merit to serve as a reference guide for the production and application of gelatin in academia and industry and will act as a platform for the development of improved methods for gelatin authentication.     

基于PCR技术的肉类成分溯源鉴定方法研究进展

近几年屡屡曝光的食品安全事故引起了社会的广泛关注,食品安全已经成为社会共同关注的问题,肉类掺假造假现象更是层出不穷,其中用低价鸡肉、鸭肉、猪肉等掺入、冒充牛羊肉成为主要的掺假方式。国内外进行肉类掺假鉴定主要以核酸作为靶标,核酸鉴定也是物种鉴别最常用、最核心的方法,以DNA检测为基础建立起来的DNA条形码、多重PCR、荧光定量PCR、荧光探针等技术也得到空前发展和广泛应用。我国针对动物源性成分检测也制定了相关国家标准和行业标准,但大多现行标准中基于DNA检测建立的PCR技术只能检测单一物种,滞后于技术的发展。目前,基于PCR发展起来的衍生技术凭借其高灵敏度、强特异性和高通量等优势在动物源性成分检测工作中显示出巨大潜力,也是肉类成分鉴定未来的重要方向。本文综述了PCR技术在肉类检测中的研究概况和现行标准的技术概况,以期为肉类成分鉴定研究提供信息。     

Meat quality evaluation by hyperspectral imaging technique: an overview

During the last two decades, a number of methods have been developed to objectively measure meat quality attributes. Hyperspectral imaging technique as one of these methods has been regarded as a smart and promising analytical tool for analyses conducted in research and industries. Recently there has been a renewed interest in using hyperspectral imaging in quality evaluation of different food products. The main inducement for developing the hyperspectral imaging system is to integrate both spectroscopy and imaging techniques in one system to make direct identification of different components and their spatial distribution in the tested product. By combining spatial and spectral details together, hyperspectral imaging has proved to be a promising technology for objective meat quality evaluation. The literature presented in this paper clearly reveals that hyperspectral imaging approaches have a huge potential for gaining rapid information about the chemical structure and related physical properties of all types of meat. In addition to its ability for effectively quantifying and characterizing quality attributes of some important visual features of meat such as color, quality grade, marbling, maturity, and texture, it is able to measure multiple chemical constituents simultaneously without monotonous sample preparation. Although this technology has not yet been sufficiently exploited in meat process and quality assessment, its potential is promising. Developing a quality evaluation system based on hyperspectral imaging technology to assess the meat quality parameters and to ensure its authentication would bring economical benefits to the meat industry by increasing consumer confidence in the quality of the meat products. This paper provides a detailed overview of the recently developed approaches and latest research efforts exerted in hyperspectral imaging technology developed for evaluating the quality of different meat products and the possibility of its widespread deployment.