荧光免疫分析法检测食品中黄曲霉毒素的研究进展

黄曲霉毒素是迄今发现的真菌毒素中毒性最强的一类,是公认的I类致癌物,在粮食、坚果、油脂、乳及其制品等多种食品中均有发现。因此,研究出准确、快速、便捷的检测方法对保障食品安全和人类健康具有重要意义。荧光免疫分析法具有灵敏度高、准确性好、检测速度快、操作简单等优势,适合现场高通量快速筛查,近年来被广泛地用于检测食品中的黄曲霉毒素。该文介绍了4类常用的荧光标记物(荧光素、量子点、上转换纳米粒子和稀土离子螯合物),并详细阐述了5种荧光免疫分析法(荧光免疫吸附法、荧光免疫层析法、荧光偏振免疫分析、荧光共振能量转移免疫分析和时间分辨免疫分析)的检测原理,以及上述方法应用于检测食品中黄曲霉毒素的国内外最新研究进展,为今后的研究提供借鉴。     

SPE法低成本快速检测乳制品中黄曲霉毒素M1

黄曲霉毒素,1993年被世界卫生组织的癌症研究机构划定为1类致癌物.其中黄曲霉毒素B1毒性和致癌性最强,而黄曲霉毒素M1是黄曲霉毒素B1的代谢物,主要存在于牛奶中.卫生部官网一份有关食物中毒的说明文件指出,黄曲霉毒素主要损伤肝脏,致癌性很强,我国乳及乳制品中规定黄曲霉毒素M1限量为0.5μg/kg. 目前检测黄曲霉毒素的方法通常为免疫亲和层析净化高效液相色谱法和酶联免疫吸附法(ELISA)这两种方法.酶联免疫吸附法(ELISA)操作简单,快速,但灵敏度低且会出现假阳性,需使用免疫亲和柱法进行精确定性、定量,而免疫亲和柱又存在使用繁琐、价格昂贵、储存条件苛刻、保存期短等缺点.     

AB SCIEX黄曲霉毒素整体解决方案——全面解决黄曲霉毒素B1、B2、G1、G2和M1的检测

日前,在液体乳产品中发现黄曲霉毒素M1超标,引发公众广泛关注.ABSCIEX公司一直致力于为客户提供全方位的解决方案,在事件爆发的第一时间建立并向客户提供测定黄曲霉毒素的整体解决方案. 黄曲霉毒素检测常见的方法为经免疫亲和柱净化后,采用HPLC(高效液相色谱法)、TLC(薄层层析法)、LCMS/MS(液质联用)及ELISA(酶联免疫)法检测.其中ELISA法使用较为普遍,但因其定量不准确,只能作为阳性样品的初筛工具;LC-MS/MS检测方法具有灵敏度高,快速、准确等特点,适用于乳制品中黄曲霉毒素的检测,被广泛应用于食品安全检测.AB SCIEX公司就是用高效液相串联质谱法建立的黄曲霉毒素整体解决方案——iMethodTM.     

黄曲霉毒素B1检测方法的研究分析

黄曲霉毒素B1是黄曲霉毒素中毒性最强、危害最大的一种。由于国内外限量标准的不统一,在出口食品中由于黄曲霉毒素不合格而被通报的食品占了相当大的比例。这也对我国检测黄曲霉毒素B1的技术提出了更高的要求,在众多检测方法中酶联免疫法也因其快速、方便等特点,逐渐被认可,成为了黄曲霉毒素B1检测工作初筛的工具。     

无需衍生化样品处理快速测定食品中黄曲霉毒素B_1

目的建立无需衍生化法.超高效液相色谱直接测定食品中的黄曲霉毒素B_1的快速检测方法。方法样品经甲醇.水(7:3,v:v)提取,免疫亲和柱净化,C_(18)色谱柱分离后,采用大体积流通池荧光检测器检测。结果黄曲霉毒素B_1在0.10~5.00 ng/mL范围内与峰面积呈良好线性关系,R~20.999,在饼干、粉丝、点心、沙琪玛、蛋糕、辣椒酱6种基质中,黄曲霉毒素B_1在0.50~2.00μg/kg范围内加标,平均回收率在71.7%~111.2%之间,相对标准偏差(RSDs)为3.8%~11.5%,检出限(LOD)为0.05 gg/kg。结论本方法灵敏、快速、无衍生、结果准确,能够满足食品中黄曲霉毒素B_1残留的检测需求。     

黄曲霉毒素M1 ELISA试剂盒的检测效果研究

目的为验证本公司生产的黄曲霉毒素M1酶联免疫法(ELISA)检测试剂盒的检测效果。方法用ELISA检测试剂盒对牛奶、酸奶、奶粉和干酪等4种样品中黄曲霉毒素M_1的残留量进行检测,并与高效液相色谱荧光检测法进行对照。结果该试剂盒对4种样品的最低检测限分别为0.039、0.192、0.306、0.199μg/kg(L),试剂盒板内板间变异系数均小于5%;对4种样品做加标回收试验,其回收率均在93%~120%之间,变异系数均小于10%;该方法与高效液相色谱法检测实际样品的阴、阳性判断结果一致。结论该方法稳定、可靠、灵敏,可满足食品中黄曲霉毒素M_1残留快速检测的需要。     

黄曲霉毒素检测方法及其应用

黄曲霉毒素(aflatoxin AFT)是由黄曲霉分泌的一种毒性极大的致癌化合物,在实际生活中黄曲霉毒素会对食物造成严重污染,因此是我国的重点检测项目,在食物流通中黄曲霉毒素会直接对人体造成严重危害,进入人体后因极强的毒性作用,可诱发人体或者动物肝癌,常引发具有极大破坏性的公共卫生案件。本篇研究针对黄曲霉毒素的检测进行了分析,经过总结之后形成了对黄曲霉毒素检测方法的综述,本文主要对薄层层析法、高效液相色谱法、微柱法以及酶联吸附法等检测方法在食品黄曲霉毒素检测中的应用进行综合分析。     

食品中黄曲霉毒素总量免疫分析方法研究进展

黄曲霉毒素(aflatoxins,AFs)污染食品的广泛性、严重性和危害性倍受食品行业的关注,单一黄曲霉毒素检测已不能满足行业的需求。针对食品黄曲霉毒素总量检测目前普遍采用理化分析方法,相比之下由于免疫分析技术具有高特异性、高灵敏性、简便性等技术优势,已成为今后的发展方向和重要的研究课题。本文就食品黄曲霉毒素总量免疫分析技术的研究进展作简要综述,旨在为食品AFs总量免疫分析提供新的方法和思路。     

基于纳米材料构建免疫传感器快速测定粮油食品中的黄曲霉毒素B_1

以改性壳聚糖和纳米金复合物为电极修饰材料,包埋固定黄曲霉毒素B_1抗体制备信号放大型免疫传感器。以K_3[Fe(CN)_6]为探针,利用循环伏安法和差分脉冲伏安法研究其免疫反应对传感器响应电流的影响,结果表明,免疫响应电流与溶液中AFB_1的浓度在0.1~1.1 ng/m L范围内成线性关系,最低检测限为0.05 ng/m L(S/N=3);免疫传感器的特异性、稳定性和重现性较好。利用该法对花生、花生油等实际样品中的黄曲霉毒素B_1进行检测,回收率在87.8~98.2%,检测精确度优于ELISA试剂盒,可用于食品中黄曲霉毒素B_1的快速检测。     

间接竞争ELISA检测试剂盒测定粮油食品中的 黄曲霉毒素B1

目的建立一套适用于食品中黄曲霉毒素B1的快速、简便、准确的检测方法,同时,进一步验证本公司研制的黄曲霉毒素ELISA检测试剂盒的检测效果。方法用酶联免疫法和高效液相色谱法分别对市售的食用油、花生、谷物样品中AFB1的污染程度进行检测分析。结果用ELISA法和HPLC法测定食用油样品的回收率分别为87.1%~92.7%和85.8%~100.8%;花生样品的回收率分别为76.0%~92.8%和76.3%~92.9%;谷物样品的回收率分别为82.6%~92.7%和82.7%~106.4%,花生加标样品6次平行测定的RSD分别为1.29%~2.46%和5.15%~8.70%,11份阳性样品测定结果表明2种方法具有很好的一致性(r2=0.995)。结论 ELISA法和HPLC法均有很好的线性关系,且测定结果相近。本公司研制的黄曲霉毒素ELISA试剂盒可以快速地检测食品中黄曲霉毒素B1,分析周期短,分析效率高。     

Current Immunoassay Methods for the Rapid Detection of Aflatoxin in Milk and Dairy Products

The presence of mycotoxins in foodstuff causes serious health problems to consumers and economically affects the food industry. Among the mycotoxins, aflatoxins are very toxic and highly carcinogenic contaminants which affect the safety of many foods, and therefore endanger human health. Aflatoxin M₁ (AFM₁) found in milk results from the biotransformation of aflatoxin B₁. Many efforts have been made to control the source of AFM₁ from farmers to dairy product companies. However, AFM₁ escapes ordinary methods of food treatment such as cooking, sterilization, and freezing, hence it appears in milk and dairy products. The presence of high levels of AFM₁ constitutes an alarming threat as milk and dairy products contain essential nutrients for human health, especially for infants and children. For this reason, there is a pressing need for developing a fast and reliable screening method for detecting trace aflatoxins in food. Several analytical methods based on high‐performance liquid chromatography (HPLC) and mass spectroscopy have been used for aflatoxin detection; however, they are expensive, time‐consuming, and require many skills. Recently, immunoassay methods, including enzyme‐linked immunosorbent assay (ELISA), immunosensors, and lateral flow immunoassay (LFIA), have been preferred for food analysis because of their improved qualities such as high sensitivity, simplicity, and capability of onsite monitoring. This paper reviews the new developments and applications of immunoassays for the rapid detection of AFM₁ in milk.     

Detection of aflatoxins in tea samples based on a class‐specific monoclonal antibody

An enzyme‐linked immunosorbent assay (ELISA) using a monoclonal antibody was established to detect aflatoxin B₁ (AFB₁) in tea. The antibody was prepared from a hybridoma derived by fusing Sp2/0‐Ag14 myeloma cells and immunised spleen B cells. The effects from pH, ionic strength, and organic solvents on immunoassay were optimised and the 50% inhibition (IC₅₀) value was 0.057 ± 0.007 ng mL^?1. Spiked black and green tea samples at 10, 20 and 50 ng g^?1 levels of AFB₁ were detected with this proposed ELISA. The recoveries for black tea samples ranged from 68.5% to 117.7% and 73.5 to 114.3% for green tea samples. This immunoassay showed no cross‐reactions with other mycotoxin family but good recognition with related aflatoxins. These results indicate that the ELISA assay could be used as a screening method for aflatoxin detection in tea samples.     

Analytical methods for the determination of zeranol residues in animal products: A review

Analytical methods for zeranol residues are reviewed. Zeranol was a widely used as an anabolic promoter, and it could give rise to very low residues in the edible tissues of food animals. Zeranol was officially banned in Europe due to safety concerns because of its potential carcinogenic and endocrine-disrupting biological activity. A few analytical methods for determination of zeranol are reported in the literature and most of the methods such as thin-layer chromatography (TLC), gas chromatography-mass spectrometry (GC/MS), high-performance liquid chromatography (HPLC), liquid chromatography-mass spectrometry (LC/MS) and immunoassay are reviewed in this paper. Specific aspects of analysing zeranol such as sample selection, sample handling, method selection and chromatographic conditions are discussed. The instrumental methods such as LC/MS and GC/MS provide sensitive and specific techniques, but are very laborious and expensive. These methods are suitable for confirmation but not for screening of large numbers of samples. A rapid, sensitive and specific assay is needed to detect positive samples in routine analysis, and immunoassay offers practical advantages over the conventional instrumental methods in rapid analysis of zeranol residues. Immunochemical methods such as enzyme-linked immunoabsorbant assay (ELISA) are simple, rapid and cost-effective, with adequate sensitivity and specificity to detect small molecules. This review can be considered as a basis for further research aimed at identifying the most efficient approaches for the analysis of zeranol.     

荧光免疫定量法即时检测谷物中黄曲霉毒素B1

目的 基于时间分辨荧光免疫层析技术,研制快速定量检测试纸条,用于粮谷物中黄曲霉毒素B1的检测。方法 以荧光微球为标记物,采用DNP独立质控体系和竞争法检测原理,构建了粮谷物中黄曲霉毒素B1荧光免疫定量即时检测（point-of-care testing ,POCT）方法。评价其准确度、重复性、特异性、与仪器确证方法符合度。结果 该方法最适条件为：pH7.8硼酸盐（BB）活化,pH7.5磷酸盐（PB）偶联,微球抗体质量比为5:4,抗原抗体质量比为50:1。检测限在0.299 ng/g -0.997 ng/g之间,定量限在0.544 ng/g -2.663 ng/g。代表性样本准确度为92.2%-111%,重复性为3.2%-7.5%。除与黄曲霉毒素B2有交叉反应（6.1%）,与其他类似物无（<0.1%）。该法检测结果与仪器确证法一致性好,在-12.5%—15.9%之间。结论 提供了一种准确、快速、定量、灵敏、便捷、适合现场检测粮谷物中黄曲霉毒素B1的时间分辨荧光免疫检测试纸条。     

Head space sensor array for the detection of aflatoxin M1 in raw ewe's milk

A novel screening method was developed for simple and rapid detection of aflatoxin M1 contamination in raw ewe's milk samples without the need for sample pretreatment. The method was based on the use of a commercial head space sensor array system constituted by 12 metal oxide semiconductor sensors, 10 metal oxide semiconductor field-effect transistor sensors, and a pattern recognition software. Twenty-four raw milk samples collected from two different groups of ewes fed with a formulated feed that contained increasing amounts of aflatoxin B1 and six noncontaminated ewe's milk samples were analyzed. The results obtained by using the head space sensor array, processed by statistical methods, made it possible to group the samples according to the presence or the absence of aflatoxin M1. Sample classification was in complete agreement with the aflatoxin M1 content measured by an enzyme-linked immunosorbent assay procedure. This is the first report, to our knowledge, of detection of aflatoxin M1 in ewe's milk by a multisensor array.     

万古霉素和去甲万古霉素检测方法研究进展

万古霉素和去甲万古霉素化学结构、药理性质和抗菌作用相似,均属于糖肽类抗生素,被广泛应用于细菌感染的治疗,尤其是一些超级细菌。临床上,药物治疗方案是依据血液中药物含量而确立的。另外药物的不合理使用会造成其在农产品中的残留,对人们的食用安全具有危害性,因此需要对农产品中药物残留进行监测,并建立高效的检测方法刻不容缓。本文对万古霉素和去甲万古霉素的检测方法进行综述,分别介绍了万古霉素在医疗卫生领域和动物源食品中不同的检测方法,包括高效液相色谱法(high performance liquid chromatography,HPLC)、液相色谱-串联质谱法(liquid chromatography-tandem mass spectrometry,LC-MS/MS)、酶放大免疫法(enzyme-multiplied immunoassay technique,EMIT)、酶联免疫法(enzyme linked immunosorbent assay,ELISA)、荧光偏振免疫法(fluorescence polarization immunoassay,FPIA)等,详细介绍了上述方法的检测原理、研究现现状及实际应用情况,并对其发展趋势进行了展望。     

粮油作物中黄曲霉毒素风险预测预警研究进展

粮油作物中黄曲霉毒素污染严重影响着人和动物的健康。开展粮油作物中黄曲霉毒素产生关键影响因素研究,构建早期风险预测模型,是减少污染发生、保障人类健康和减少经济损失的重要途径。本文在总结黄曲霉毒素在粮油作物中风险发生的影响因素的基础上,从黄曲霉毒素易发多发的环节如作物收获期和储藏期讨论了预测模型建的研究进展,同时归纳了目前预测模型主要建立方法,包括基于回归、神经网络等方法建立的数据驱动模型和基于黄曲霉毒素产生机制建立的机理模型。通过分析目前的研究,讨论了未来粮油作物中黄曲霉毒素风险预测模型的研究方向和应用场景,为开展我国作物中黄曲霉毒素污染风险早期预测和污染防控研究提供依据。     

Detection of aflatoxin-producing molds in Korean fermented foods and grains by multiplex PCR

An assay based on multiplex PCR was applied for the detection of potential aflatoxin-producing molds in Korean fermented foods and grains. Three genes, avfA, omtA, and ver-1, coding for key enzymes in aflatoxin biosynthesis, were used as aflatoxin-detecting target genes in multiplex PCR. DNA extracted from Aspergillus flavus, Aspergillus parasiticus, Aspergillus oryzae, Aspergillus niger, Aspergillus terreus, Penicillium expansum, and Fusarium verticillioides was used as PCR template to test specificity of the multiplex PCR assay. Positive results were achieved only with DNA that was extracted from the aflatoxigenic molds A. flavus and A. parasiticus in all three primer pairs. This result was supported by aflatoxin detection with direct competitive enzyme-linked immunosorbent assay (DC-ELISA). The PCR assay required just a few hours, enabling rapid and simultaneous detection of many samples at a low cost. A total of 22 Meju samples, 24 Doenjang samples, and 10 barley samples commercially obtained in Korea were analyzed. The DC-ELISA assay for aflatoxin detection gave negative results for all samples, whereas the PCR-based method gave positive results for 1 of 22 Meju samples and 2 of 10 barley samples. After incubation of the positive samples with malt extract agar, DC-ELISA also gave positive results for aflatoxin detection. All Doenjang samples were negative by multiplex PCR and DC-ELISA assay, suggesting that aflatoxin contamination and the presence of aflatoxin-producing molds in Doenjang are probably low.     

贝类毒素大田软海绵酸的免疫分析检测技术研究进展

大田软海绵酸(okadaicacid,OA)是一种广泛存在于贝类等生物中的海洋生物毒素,可引起人或动物的急性中毒,对食品安全和海产养殖具有严重危害。因此建立快速、可靠、灵敏的OA检测技术具有重要意义。免疫分析检测技术基于抗原抗体的结合,特异性强、灵敏度高、应用范围广,是当前检测贝类毒素OA的主要手段。本文综述了近年来针对贝类毒素OA的免疫分析检测技术,其中包括酶联免疫吸附检测、免疫层析检测、时间分辨荧光免疫检测和基于免疫传感器的检测技术等。本文着重阐述了不同免疫分析技术的原理及其在OA检测中的实际应用,同时探讨了免疫分析技术在贝类毒素OA检测方面的挑战和发展趋势,以期为开发性能更加优异的OA免疫检测技术提供研究思路。