發(fā)布時間：2018-01-11 03:19

  本文關(guān)鍵詞：羊肉及其制品中摻假動物源性成分?jǐn)?shù)字PCR技術(shù)精準(zhǔn)定量研究　出處：《中國農(nóng)業(yè)大學(xué)》2017年博士論文　論文類型：學(xué)位論文

  更多相關(guān)文章： 數(shù)字PCR 羊肉 精準(zhǔn)定量 真?zhèn)舞b別 摻假

【摘要】：近年來,食品欺詐問題日益突出,肉制品摻假現(xiàn)象尤為嚴(yán)重。前幾年歐洲爆發(fā)的"馬肉風(fēng)波"揭露了多國肉制品的摻假現(xiàn)象;我國同樣存在不法分子將廉價肉摻入高價肉中,以獲取巨額利潤的經(jīng)濟(jì)利益驅(qū)動型摻假現(xiàn)象。在肉類產(chǎn)業(yè)中,肉羊是我國畜牧業(yè)的重要支柱產(chǎn)業(yè)之一,消費(fèi)需求旺盛。然而,用雞、鴨和豬肉摻入甚至替代羊肉的事件屢見不鮮。這些問題的出現(xiàn)嚴(yán)重干擾了我國畜肉產(chǎn)業(yè),制約了肉品質(zhì)量的提升,并嚴(yán)重?fù)p害了消費(fèi)者的權(quán)益,甚至涉及宗教信仰等問題。因此,提升羊肉的真?zhèn)舞b別能力對于保障居民的食肉安全有著重要的現(xiàn)實(shí)意義。目前,羊肉的真?zhèn)舞b別檢測大多停留在定性和半定量階段,目前尚無精準(zhǔn)的定量檢測方法,因此無法確定摻假成分是有意添加還是無意污染,更無法作為食品摻假犯罪中量刑依據(jù)的定量檢測標(biāo)準(zhǔn)方法推廣。因此,亟待建立一種精準(zhǔn)、快速、高通量的定量方法,對羊肉及其加工品中不同動物源性摻假成分進(jìn)行精準(zhǔn)定量檢測。本論文基于微滴式數(shù)字PCR技術(shù),建立了針對羊肉及其多種動物源性摻假成分的定性及精準(zhǔn)定量檢測方法,具體研究工作如下:1.針對羊肉及其摻假物的定性檢測,選取單拷貝核基因作為檢測靶標(biāo),通過篩選和優(yōu)化各物種的特異性引物、探針和數(shù)字PCR體系,建立了基于數(shù)字PCR技術(shù)的羊肉及其摻假物定性檢測方法;對方法特異性、穩(wěn)定性以及靈敏度的驗(yàn)證結(jié)果表明該檢測體系可以對摻假物種含量為0.1%的羊肉樣品實(shí)現(xiàn)穩(wěn)定檢測,且靈敏度高、特異性好。2.針對純?nèi)庵破返木珳?zhǔn)定量,通過理論推導(dǎo)獲得單位質(zhì)量兩種肉基因拷貝數(shù)之比的倍增系數(shù),并進(jìn)行實(shí)驗(yàn)驗(yàn)證,據(jù)此將羊肉和其他摻假肉的基因拷貝數(shù)轉(zhuǎn)化為質(zhì)量分?jǐn)?shù),最低定量檢測限達(dá)到1%(w/w),模擬不同條件對肉制品進(jìn)行處理,結(jié)果表明定量方法的準(zhǔn)確度和重復(fù)性不受樣品部位、超高壓、加熱和冷凍處理等條件的影響。3.針對含有食品配料和添加劑羊肉制品的精準(zhǔn)定量,通過在樣品中加入內(nèi)標(biāo)(火雞)肉種,制備標(biāo)準(zhǔn)樣品,繪制標(biāo)準(zhǔn)曲線,計(jì)算得到各物種基因拷貝數(shù)與質(zhì)量的轉(zhuǎn)化公式,進(jìn)而計(jì)算出肉的質(zhì)量,最低定量檢測限為1%(w/w),模擬不同條件對肉制品進(jìn)行處理,結(jié)果表明定量方法的準(zhǔn)確度和重復(fù)性不受添加劑和配料的種類、樣品部位、超高壓、加熱和冷凍處理?xiàng)l件的影響。4.針對羊肉中同時摻雜多種動物源性成分情況下的定量檢測,在單一物種定量檢測方法的基礎(chǔ)上,建立基于倍增系數(shù)法的多重?cái)?shù)字PCR定量檢測體系,包括羊肉中同時摻雜雞、鴨和豬肉的四重?cái)?shù)字PCR定量檢測體系和羊肉中同時摻雜狐貍、水貂和貉肉的四重?cái)?shù)字PCR同時定量檢測體系;通過在混合樣品中加入內(nèi)標(biāo)(火雞),計(jì)算摻假物種與內(nèi)標(biāo)肉種的比值,建立了基于內(nèi)標(biāo)法的五重?cái)?shù)字PCR定量檢測體系。綜上所述,本論文基于微滴式數(shù)字PCR技術(shù)建立的兩種肉制品中物種成分含量分析方法,可分別實(shí)現(xiàn)對羊肉中多種摻假物種的定性篩查、單一摻假成分的精準(zhǔn)定量檢測以及多種動物源性成分同時精準(zhǔn)定量檢測,從而初步建立了羊肉質(zhì)量檢測的表征屬性識別技術(shù)體系,該體系不僅可以定性和定量羊肉中摻雜的動物源性成分,也可以應(yīng)用于其他肉類及其他食品摻假的定性和定量檢測,為羊肉及其制品的市場監(jiān)管和相關(guān)執(zhí)法提供有力的技術(shù)保障。
[Abstract]:In recent years, food fraud problems have become increasingly prominent, meat adulteration phenomenon is particularly serious. A few years ago the European outbreak of "horsemeat" reveals the phenomenon of adulterated multinational meat products; China also has the criminals will be cheap meat meat mixed with high price, in order to obtain huge profits driven by economic interests in the adulteration. The meat industry, meat is China's animal husbandry is one of the important pillar industry, consumer demand. However, with chicken, duck and pork mixed with mutton or even replace the event. It is often seen. these problems serious interference in the meat industry, restricting the meat quality improvement, and serious damage to the interests of consumers and even religion and other issues. Therefore, enhance the ability to identify the authenticity of mutton has important practical significance for the protection of residents food safety. At present, the authenticity of mutton are qualitative detection Semi quantitative and quantitative detection method, there is no accurate current, therefore unable to determine the adulteration is intentionally added or not pollution, not as the extension of the standard method for quantitative detection of adulteration of food crime sentencing basis. Therefore, urgent need to establish an accurate, rapid, high-throughput quantitative method, accurate quantitative detection of different animal the source of mutton and its processed products of adulteration. The droplet digital PCR technology based on the established detection method for qualitative and a variety of animal derived mutton adulteration and accurate quantitative, specific research work is as follows: 1. for mutton and adulterated qualitative detection, selection of single copy nuclear gene as target detection. Through the screening and optimization of specific primers for each species of the probe and the digital PCR system was developed to detect mutton and its adulterant qualitative digital PCR technology based on; The specificity of the method, stability and sensitivity of the verification results show that the detection system of adulteration species content of 0.1% mutton samples to achieve stable detection, high sensitivity, good specificity and accurate quantitative.2. for pure meat products, obtain copies of unit quality of two kinds of meat gene of the ratio of the number of multiplication coefficient by theoretical derivation, and based on the experiment, mutton and other meat adulteration gene copy number into the mass fraction, the minimum quantitative detection limit reached 1% (w/w), the simulation of different conditions on meat processing, results showed that the accuracy and repeatability of quantitative method from the sample site, ultra high pressure, heating and freezing conditions effect of.3. for accurate quantification of containing food ingredients and additives of mutton products, in the sample by adding internal standard (Turkey) meat, the preparation of standard sample, drawing the standard curve, the calculated A gene copy number and mass conversion formula, and then calculate the meat quality, the lowest limit of quantification was 1% (w/w), the simulation of different conditions on meat processing, the result show that the type, accuracy and repeatability of quantitative methods without additives and ingredients sample parts, high pressure, heating and freezing treatment the effect of.4. on various conditions of quantitative animal derived ingredients and mutton in the doping test, based on the quantitative detection method of single species, the establishment of multiple digital PCR quantitative detection system based on the multiplier method, including sheep meat and chicken and the fox four doped, doped heavy digital PCR quantitative detection system and mutton duck and pork, four mink and racoon dog meat and digital PCR quantitative detection system; through the mixed samples with internal standard (Turkey), calculation of species and the internal standard of meat adulteration ratio value is established based on Five digital PCR quantitative detection system of internal standard method. To sum up, this paper analysis of species composition content of two kinds of meat products to establish the droplet digital PCR technology based on can be realized respectively for qualitative screening multiple species adulteration in Mutton adulteration, single precision quantitative detection and a variety of animal derived ingredients at the same time accurate quantitative detection, we preliminarily established the system characterization of attribute recognition technology of mutton quality detection, the system can not only components of animal origin doping of lamb in the qualitative and quantitative, qualitative and quantitative detection can also be applied to other meat and other food adulteration, to provide strong technical support for mutton and its products and related market supervision law enforcement.

【學(xué)位授予單位】：中國農(nóng)業(yè)大學(xué)
【學(xué)位級別】：博士
【學(xué)位授予年份】：2017
【分類號】：TS251.7

【參考文獻(xiàn)】

相關(guān)期刊論文 前10條

1 周廣運(yùn);王桂姬;任皓威;蘆乾;楊艷;郭立海;劉寧;;超高效液相色譜-串聯(lián)質(zhì)譜法鑒定豬肉的特異性肽生物標(biāo)志物[J];分析化學(xué);2017年02期

2 陳加齊;魏曉娟;朱增勇;趙安平;;近年來全球畜產(chǎn)品消費(fèi)趨勢分析及未來展望[J];農(nóng)業(yè)展望;2017年01期

3 段敏;李強(qiáng);戴岳;劉鵬;郝佳;王彥蓉;劉文;;肉制品質(zhì)量安全風(fēng)險(xiǎn)因子檢測與控制研究進(jìn)展[J];肉類工業(yè);2017年01期

4 馮兆民;舒躍龍;;數(shù)字PCR技術(shù)及其應(yīng)用進(jìn)展[J];病毒學(xué)報(bào);2017年01期

5 鄔升超;黃斐;趙瀛;虞倩;韓序;王蓓麗;張春燕;郭瑋;樓文暉;潘柏申;;利用多重?cái)?shù)字PCR檢測KRAS相關(guān)突變[J];復(fù)旦學(xué)報(bào)(醫(yī)學(xué)版);2016年06期

6 張穎穎;趙文濤;李慧晨;馬燕紅;郭文萍;李瑩瑩;;液相色譜串聯(lián)質(zhì)譜對摻假牛肉的鑒別及定量研究[J];現(xiàn)代食品科技;2017年02期

7 張秋;肖平輝;;從“僵尸肉”事件談肉制品安全風(fēng)險(xiǎn)管理[J];肉類研究;2016年10期

8 金萍;結(jié)莉;陸俊;陳英;丁洪流;;Taqman探針熒光聚合酶鏈?zhǔn)椒磻?yīng)實(shí)時同步鑒定動物源性食品中豬肉、雞肉源性成分[J];肉類研究;2016年09期

9 唐穗平;張燕;黃景輝;;廣東省牛羊肉及其制品中摻雜摻假情況的調(diào)查分析[J];食品安全質(zhì)量檢測學(xué)報(bào);2016年05期

10 于曉帆;高宏偉;孫敏;肖西志;李榮貴;;熒光PCR和數(shù)字PCR法檢測轉(zhuǎn)基因DAS-44406-6品系大豆[J];食品科學(xué);2016年16期

相關(guān)重要報(bào)紙文章 前2條

1 李俊;;業(yè)內(nèi)呼吁：加強(qiáng)對原料肉和標(biāo)簽標(biāo)注的監(jiān)管[N];中國食品報(bào);2017年

2 陳穎;;食品打假鑒偽亟須插上科技翅膀[N];科技日報(bào);2015年

相關(guān)碩士學(xué)位論文 前2條

1 董洋洋;實(shí)時熒光PCR對牛肉中摻入鴨肉和豬肉的定量檢測研究[D];黑龍江八一農(nóng)墾大學(xué);2016年

2 呂二盼;動物源性食品中各種動物源性肉及肉制品鑒別檢驗(yàn)的研究[D];河北農(nóng)業(yè)大學(xué);2012年

，

本文編號：1407921