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用于食品中榛子过敏原PCR扩增检测的一次性电化学DNA阵列。

Disposable electrochemical DNA-array for PCR amplified detection of hazelnut allergens in foodstuffs.

作者信息

Bettazzi Francesca, Lucarelli Fausto, Palchetti Ilaria, Berti Francesca, Marrazza Giovanna, Mascini Marco

机构信息

Dipartimento di Chimica, Università degli Studi di Firenze, Via della Lastruccia 3, 50019 Sesto Fiorentino (Fi), Italy.

出版信息

Anal Chim Acta. 2008 Apr 28;614(1):93-102. doi: 10.1016/j.aca.2008.03.027. Epub 2008 Mar 19.

Abstract

An electrochemical low-density DNA-array has been designed and implemented to be used in combination with polymerase chain reaction (PCR) in order to investigate the presence of hazelnut major allergens (Cor a 1.04, Cor a 1.03) in foodstuff. Unmodified PCR products were captured at the sensor interface via sandwich hybridization with surface-tethered probes and biotinylated signalling probes. The resulting biotinylated hybrids were coupled with a streptavidin-alkaline phosphatase conjugate and then exposed to a alpha-naphthyl phosphate solution. Differential pulse voltammetry was finally used to detect the alpha-naphthol signal. The detection limits for Cor a 1.03 and Cor a 1.04 were 0.3 and 0.1 nmol L(-1), respectively (R.S.D. 10%). The optimized conditions were used to test several commercially available foodstuffs, claiming to contain or not the targeted nuts. The results were compared with those obtained with classical ELISA tests.

摘要

一种电化学低密度DNA阵列已被设计并实现,用于与聚合酶链反应(PCR)结合使用,以检测食品中榛子主要过敏原(Cor a 1.04、Cor a 1.03)的存在。未修饰的PCR产物通过与表面连接的探针和生物素化信号探针进行夹心杂交,捕获在传感器界面上。所得的生物素化杂交体与链霉亲和素-碱性磷酸酶缀合物偶联,然后暴露于α-萘基磷酸溶液中。最后使用差分脉冲伏安法检测α-萘酚信号。Cor a 1.03和Cor a 1.04的检测限分别为0.3和0.1 nmol L(-1)(相对标准偏差10%)。使用优化条件测试了几种声称含有或不含有目标坚果的市售食品。将结果与经典ELISA测试获得的结果进行了比较。

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