一种检测咖啡豆中黑曲霉的分子方法。
A molecular method for detection of Aspergillus carbonarius in coffee beans.
作者信息
Fungaro Maria Helena Pelegrinelli, Vissotto Patrícia Cristina, Sartori Danieli, Vilas-Boas Laurival Antônio, Furlaneto Márcia Cristina, Taniwaki Marta Hiromi
机构信息
Centro de Ciências Biológicas, Universidade Estadual de Londrina, P.O. Box 6001, 86051-990 Londrina-PR, Brazil.
出版信息
Curr Microbiol. 2004 Aug;49(2):123-7. doi: 10.1007/s00284-004-4273-z.
Ochratoxin A (OTA) is a carcinogenic and nephrotoxic mycotoxin that has been detected in a variety of food products, including green coffee beans. About 80% of Aspergillus carbonarius strains collected from coffee beans are able to produce OTA on this substrate. The rapid identification of this fungal species would be desirable. RAPD assays were applied to identify amplification products specific for A. carbonarius. One selected fragment, denoted OPX7809, was cloned and sequenced. Based on the nucleotide sequence obtained, specific oligonucleotides (OPX7F809 and OPX7R809) were designed and used as primers for DNA amplification. One amplified band of 809 bp was obtained from A. carbonarius genomic DNA, whereas no amplified fragment from DNA of other Aspergillus species was detected. This PCR analysis was also successfully employed to detect A. carbonarius in coffee beans. This PCR assay could contribute to the early and rapid detection of the potential presence of OTA in coffee samples.
赭曲霉毒素A（OTA）是一种具有致癌性和肾毒性的霉菌毒素，已在包括生咖啡豆在内的多种食品中被检测到。从咖啡豆中分离得到的大约80%的炭黑曲霉菌株能够在该底物上产生OTA。快速鉴定这种真菌物种是很有必要的。随机扩增多态性DNA（RAPD）分析被用于鉴定炭黑曲霉特异性的扩增产物。一个选定的片段，命名为OPX7809，被克隆并测序。基于所获得的核苷酸序列，设计了特异性寡核苷酸（OPX7F809和OPX7R809）并用作DNA扩增的引物。从炭黑曲霉基因组DNA中获得了一条809 bp的扩增带，而未检测到其他曲霉属物种DNA的扩增片段。这种PCR分析也成功地用于检测咖啡豆中的炭黑曲霉。这种PCR检测方法有助于早期快速检测咖啡样品中OTA的潜在存在情况。
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