Development and validation of a SYBR-Green I Real-Time PCR test to detect bivalves including Mytilus species in foods

Sara Graziano; Mariolina Gulli; Nelson Marmiroli

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Development and validation of a SYBR-Green I Real-Time PCR test to detect bivalves including Mytilus species in foods

机译：SYBR-Green I实时PCR测试的开发和验证，以检测食品中贻贝种类的纤维

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The incidence of allergy to seafood, and in particular to molluscs is second only to that of nuts. To protect consumers, the regulators of food products insisted on identifying molluscs as allergens. The aim was to develop quantitative assay for the presence Mytilus species in processed food products. The chosen platform was real-time PCR (qPCR) targeting either the gene encoding mitochondrial cytochrome C oxi-dase I or the nuclear gene encoding p-actin. Recombinant plasmids containing each of target regions were used as a reference for quantification purposes. Limit of detection (LOD) and of quantification (LOQ) were determined. Spiked food samples containing 50-500 μg g~(-1) of Mytilus chilensis were analysed both by qPCR and by ELISA. The former assay gave a positive outcome over this range, whereas the latter was sensitive down to a concentration of 125 μg g~(-1).

机译：对海鲜过敏的发病率，特别是软体动物的发病率仅为螺母的第二个。为了保护消费者，食品的监管机构坚持认为将软体动物鉴定为过敏原。目的是为加工食品中的存在霉菌物种进行定量测定。所选择的平台是实时PCR（QPCR），其靶向编码线粒体细胞色素C氧化酶I或编码P-肌动蛋白的核基因的基因。含有每种靶区域的重组质粒用作定量目的的参考。确定检测限（LOD）和定量（LOQ）的限制。通过QPCR和ELISA分析含有50-500μgg〜（-1）米干酪氏菌的尖刺食品样品。前者测定在该范围内产生了阳性结果，而后者敏感至125μgg〜（-1）的浓度。

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《International journal of food science & technology》 |2017年第7期|1567-1575|共9页
作者
Sara Graziano; Mariolina Gulli; Nelson Marmiroli;
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Dipartimento di Bioscienze Universita degli Studi di Parma Parco Area delle Scienze 11a Parma 43124 Italy;

Dipartimento di Bioscienze Universita degli Studi di Parma Parco Area delle Scienze 11a Parma 43124 Italy Centro Interdipartimentale SITEIA.PARMA Universita degli Studi di Parma Parco Area delle Scienze 181/A Parma 43124 Italy;

Dipartimento di Bioscienze Universita degli Studi di Parma Parco Area delle Scienze 11a Parma 43124 Italy Centro Interdipartimentale SITEIA.PARMA Universita degli Studi di Parma Parco Area delle Scienze 181/A Parma 43124 Italy;

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正文语种 eng
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Allergens; ELISA; PCR; seafood;

机译：过敏原;ELISA;PCR;海鲜;

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3. Development and Validation of a Duplex Real-Time PCR Method To Simultaneously Detect Potentially Allergenic Sesame and Hazelnut in Food [J] . KERSTIN SCHORINGHUMER, GERDA REDL, MARGIT CICHNA-MARKL Journal of Agricultural and Food Chemistry . 2009,第6期

机译：同时检测食品中潜在致敏芝麻和榛子的双重实时PCR方法的开发和验证
4. REAL-TIME PCR BASED FOOD PATHOGEN DETECTION ON A CENTRIFUGAL MICROFLUIDIC FOIL DISK INCLUDING POSITIVE- AND NO-TEMPLATE-CONTROLS [C] . Oliver Strohmeier, Nico Marquart, Daniel Mark, International Conference on Miniaturized Systems for Chemistry and Life Sciences . 2011

机译：离心微流体箔盘的实时PCR基食物病原体检测，包括阳性和无模板对照
5. Development, validation, and evaluation of a continuous, real-time, bivalve biomonitoring system. [D] . Allen, H. Joel. 2002

机译：开发，验证和评估连续，实时，双瓣生物监测系统。
6. Ecotoxicological Evaluation of the Immunocompetence of two Bivalves Species (Mya Arenaria and Mytilus Edulis) in the Saguenay Fjord Including a Salinity Gradient [O] . A. Beaudry, E. Lacaze, A. Jobin-Piché, 2015

机译：盐度梯度在萨格奈峡湾中两个双壳类（Mya Arenaria和Mytilus Edulis）免疫能力的生态毒理学评估。
7. INTRODUCTION After fourteen years of pioneering systematic work spanning from 1996 to 2010, carried out in the territory of Santa Catarina State, SC, within the Brazilian Southern region, framed in the socalled Atlantic Slope of the Southern Cone of South America (Agudo-Padrón, 2008a), a basic list of continental (land and freshwater) and marine mollusc species was compiled. Besides constant interactions and consultations with numerous national and international specialists, such a list was mostly based on available literature and/or analyses of voucher specimens deposited in collections belonging to research centers or environmental education institutes. To date (up to the first semester of 2010), this list comprises a total of 878 taxa (species and subspecies, including 695 marine and 183 continental forms), and these numbers are likely to increase as field surveys ensue. In the present study, results obtained from the author’s active participation in three recent regional field sampling expeditions dealing with marine and continental mollusc taxa, are reported. I. Official State program for listing and control of invasive exotic species Starting from November 2009, and for the first time in the history of Santa Catarina State, the presence of invasive allochthonous mollusc species in Santa Catarina State was studied and discussed through the organisation of seminars by the Official Foundation for the Environment of the State of Santa Catarina (Fundação do Meio Ambiente – FATMA) jointly with the Hórus Institute for Development and Environmental Conservation (Instituto Hórus de Desenvolvimento e Conservação Ambiental), with the main goal to compile the Official State List of Species. To date, the occurrence of a total of twenty allochthonous (exotic) forms of mollusc species has been confirmed, 14 Gastropoda and 6 Bivalvia namely, 11 terrestrial gastropods, 5 freshwater taxa (3 gastropods and 2 bivalves) and 4 marine bivalves. Taking into account the contributions of Agudo Bleicker (2006a), Agudo-Padrón (2008b) and Agudo-Padrón Lenhard (2010), the slug Pallifera sp. - the taxonomic determination of which is still in process (Thomé et al., 2006) - was included within such a list. Of these twenty Mollusca and environmental conservation in Santa Catarina State (SC, Southern Brazil): current situation [O] . A. Ignacio Agudo-Padrón 2011

机译：引言经过十四年的开展系统工作，从1996年到2010年，在南卡罗来纳州南卡罗来纳州圣卡塔琳娜州境内开展，在南美洲南锥体所谓的大西洋坡（agudo-padrón， 2008a），编制了大陆（陆地和淡水）和海洋软体动物物种的基本清单。除了与众多国家和国际专家不断互动和协商之外，这样的清单主要基于现有文献和/或存放在属于研究中心或环境教育机构的馆藏中的凭证标本的分析。到目前为止（截至2010年第一季度），该清单共包括878个分类群（物种和亚种，包括695种海洋和183种大陆形式），随着实地调查的发生，这些数字可能会增加。在本研究中，报告了作者积极参与最近三次涉及海洋和大陆软体动物类群的区域野外抽样考察的结果。 I.列入和控制外来入侵物种的官方国家方案从2009年11月开始，圣卡塔琳娜州历史上第一次通过组织“圣卡塔琳娜州”对圣卡塔琳娜州的入侵性异地软体动物物种的存在进行了研究和讨论。由圣卡塔琳娜州官方基金会（Fundaçãomomeio ambiente - FaTma）与Hórus发展与环境保护研究所（InstitutoHórusdeDesenvolvimento eConservaçãoambiental）联合举办的研讨会，主要目标是编制官方州物种清单。到目前为止，已确认共发生了20种异地（外来）形式的软体动物物种，14种腹足纲和6种双壳纲即11种陆生腹足类动物，5种淡水类群（3种腹足动物和2种双壳类动物）和4种海洋双壳类动物。考虑到agudo＆Bleicker（2006a），agudo-padrón（2008b）和agudo-padrón＆Lenhard（2010）的贡献，pallifera sp。 - 其中的分类学决定仍在进行中（Thomé等，2006） - 被列入此类清单。在圣卡塔琳娜州（sC，巴西南部）的这20个软体动物和环境保护中：当前的情况

Development and validation of a SYBR-Green I Real-Time PCR test to detect bivalves including Mytilus species in foods

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