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建立并验证了一种免疫层析法,可快速检测谷物样品中的伏马菌素 B1。

Development and validation of an immunochromatographic assay for rapid detection of fumonisin B1 from cereal samples.

机构信息

DRDO-BU-Center for Life Sciences, Bharathiar University Campus, Coimbatore, Tamilnadu 641046 India ; DFRL, Mysore, 11 India.

Department of Applied Nutrition, Defence Food research Laboratory, Siddarthanagar, Mysore, 570011 Karnataka India.

出版信息

J Food Sci Technol. 2014 Sep;51(9):1920-8. doi: 10.1007/s13197-013-1254-x. Epub 2014 Jan 7.

Abstract

Fumonisins are one of the most agriculturally significant environmental toxins produced by Fusarium and Aspergillus species that grow on agricultural commodities in the field or during storage. Cereals contaminated with fumonisins causes serious loss to agricultural produce leads to health problems in humans and other farm animals. In the present study, polyclonal hyperimmune sera was raised against FB1 in rabbits immunized with FB1-keyhole limpet haemocyanin (KLH). Purified antibodies were used to establish a sensitive gold nanoparticle based immunochromatographic strip (ICG) for detecting FB1 levels in cereal grains. Effective on-site detection of FB1 was achieved by developing a rapid and sensitive pAb based ICG strip. This strip had a detection limit of 5 ng mL(-1) for FB1 in cereal samples and it could be completed within 3 min. Close examination of 150 cereal samples by ICG strip method revealed that 77 were fumonisin-positive. Results obtained by the developed method was further validated with well standardized HPLC method and results of strip method was correlated well with those obtained by HPLC method. In conclusion, the developed method was a better alternative for onsite detection of FB1 in cereal samples intended for human consumption to reduce risk of humans and other farm animals. The high level of FB1 concentrations recorded in present study warrants the need to develop an awareness creation programme to the farmers of India for safe handling of cereal grains at the time of harvesting and storage of grains.

摘要

伏马菌素是由生长在田间或储存过程中农作物上的镰刀菌和曲霉菌产生的对农业生产具有重大影响的环境毒素之一。被伏马菌素污染的谷物会导致农产品严重损失,并引发人类和其他农场动物的健康问题。在本研究中,用 FB1 与血蓝蛋白(KLH)偶联物免疫家兔制备了针对 FB1 的多克隆高效价免疫血清。纯化的抗体用于建立一种基于金纳米颗粒的敏感免疫层析条(ICG),用于检测谷物中的 FB1 水平。通过开发一种快速灵敏的基于 pAb 的 ICG 条,实现了对 FB1 的有效现场检测。该条在谷物样品中对 FB1 的检测限为 5ng/mL,可在 3 分钟内完成。用 ICG 条法对 150 个谷物样品进行仔细检查,发现 77 个样品为伏马菌素阳性。用开发的方法获得的结果与经过充分标准化的 HPLC 方法进行了进一步验证,条带法的结果与 HPLC 方法的结果相关性良好。总之,该方法是一种用于检测人类食用谷物中 FB1 的替代方法,可以降低人类和其他农场动物的风险。本研究中记录的高浓度 FB1 需要在印度农民中开展安全处理谷物的宣传计划,以确保在收获和储存谷物时的安全处理。

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