Real Time PCR to detect hazelnut allergen coding sequences in processed foods

被引：37

作者：

Iniesto, Elisa ^{[1
]}

Jimenez, Ana ^{[1
]}

Prieto, Nuria ^{[1
]}

Cabanillas, Beatriz ^{[3
]}

Burbano, Carmen ^{[2
]}

Pedrosa, Mercedes M. ^{[2
]}

Rodriguez, Julia ^{[3
]}

Muzquiz, Mercedes ^{[2
]}

Crespo, Jesus F. ^{[3
]}

Cuadrado, Carmen ^{[2
]}

Linacero, Rosario ^{[1
]}

机构：

[1] Univ Complutense Madrid, Fac Biol, Dept Genet, E-28040 Madrid, Spain

[2] SGIT INIA, Dept Tecnol Alimentos, Madrid 28040, Spain

[3] Hosp 12 Octubre I 12, Inst Invest, Serv Alergia, Madrid 28041, Spain

来源：

FOOD CHEMISTRY | 2013年 / 138卷 / 2-3期

关键词：

Allergen detection; Hazelnut; High hydrostatic pressure; Processed foods; Real time-PCR; CHAIN-REACTION PCR; SANDWICH-ELISA; IDENTIFICATION; PRESSURE; RESIDUES; CLONING;

D O I：

10.1016/j.foodchem.2012.11.036

中图分类号：

O69 [应用化学];

学科分类号：

081704 ;

摘要：

A quantitative RT-PCR method, employing novel primer sets designed on Cor a 9, Cor a 11 and Cor a 13 allergen-coding sequences has been setup and validated. Its specificity, sensitivity and applicability have been compared. The effect of processing on detectability of these hazelnut targets in complex food matrices was also studied. The DNA extraction method based on (TAB-phenol-chloroform was the best for hazelnut. RT-PCR using primers for Cor a 9, 11 and 13 allowed a specific and accurate amplification of these sequences. The limit of detection was 1 ppm of raw hazelnut. The method sensitivity and robustness were confirmed with spiked samples. Thermal treatments (roasting and autoclaving) reduced yield and amplificability of hazelnut DNA, however, high-hydrostatic pressure did not affect. Compared with an ELISA assay, this RT-PCR showed higher sensitivity to detected hazelnut traces in commercial food-stuffs. The RT-PCR method described is the most sensitive of those reported for the detection of hazelnut traces in processed foods. (C) 2012 Elsevier Ltd. All rights reserved.

引用

页码：1976 / 1981

页数：6

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