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通过聚合酶链式反应(PCR)分析鉴别黑曲霉及属于黑曲霉组的其他曲霉属菌种。

Discrimination of Aspergillus niger and other Aspergillus species belonging to section Nigri by PCR assays.

作者信息

González-Salgado Amaia, Patiño Belén, Vázquez Covadonga, González-Jaén M Teresa

机构信息

Departamento de Genética, Facultad de Biología, Universidad Complutense de Madrid, José Antonio Nováis 2, 28040-Madrid, Spain.

出版信息

FEMS Microbiol Lett. 2005 Apr 15;245(2):353-61. doi: 10.1016/j.femsle.2005.03.023.

Abstract

Aspergillus species included in section Nigri are common in plant products and processed food, such as grapes, cereals, coffee and derivatives, particularly in warm and tropical climates. Two of these species, A. carbonarius and A. niger, are known to produce ochratoxin A (OTA), a potent nephrotoxin and carcinogenic to human (group 2B). Recognition of the several species of this section is difficult and requires considerable expertise using conventional methods based on morphological features. In this work we describe rapid, sensitive and robust assays based on the PCR technique to discriminate the main species included in section Nigri: A. japonicus, A. heteromorphus, A. ellipticus and the two morphologically indistinguishable species of the A. niger aggregate: A. niger and A. tubingensis. The species-specific primers have been designed on the basis of ITS (internal transcribed spacers of rDNA units) sequence comparisons obtained from several Aspergillus strains and have been tested in a number of strains from different origins and hosts. These PCR assays, based on multi-copy sequences, are highly sensitive and specific and represent a good tool for an early detection of OTA-producing Aspergillus species in order to prevent OTA from entering the food chain.

摘要

黑曲霉组中的曲霉物种在植物产品和加工食品中很常见,如葡萄、谷物、咖啡及其衍生物,在温暖和热带气候地区尤为如此。其中的两个物种,即炭黑曲霉和黑曲霉,已知会产生赭曲霉毒素A(OTA),这是一种强效肾毒素,对人类具有致癌性(2B类)。识别该组中的多个物种很困难,需要运用基于形态特征的传统方法的相当专业知识。在这项工作中,我们描述了基于PCR技术的快速、灵敏且可靠的检测方法,以区分黑曲霉组中的主要物种:日本曲霉、异形曲霉、椭圆曲霉以及黑曲霉复合体中形态上无法区分的两个物种:黑曲霉和管囊曲霉。物种特异性引物是根据从多个曲霉菌株获得的ITS(核糖体DNA单位的内部转录间隔区)序列比较设计的,并已在来自不同来源和宿主的多个菌株中进行了测试。这些基于多拷贝序列的PCR检测方法高度灵敏且特异,是早期检测产OTA曲霉菌种的良好工具,以便防止OTA进入食物链。

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