通过聚合酶链反应检测和定量生咖啡中的赭曲霉。
Detection and quantification of Aspergillus ochraceus in green coffee by PCR.
作者信息
Schmidt H, Bannier M, Vogel R F, Niessen L
机构信息
TU München, Lehrstuhl für Technische Mikrobiologie, Freising, Germany.
出版信息
Lett Appl Microbiol. 2004;38(6):464-9. doi: 10.1111/j.1472-765X.2004.01524.x.
AIMS
The aim of this study was to detect and quantify DNA of the ochratoxinogenic fungus Aspergillus ochraceus in green coffee and to compare the results with the ochratoxin A content of naturally contaminated samples.
METHODS AND RESULTS
A DNA extraction protocol based on a combination of ultrasonification and a commercial kit was tested for the recovery of fungal DNA. PCR and real-time PCR protocols were established for the detection of A. ochraceus. Sensitivity of the PCR was checked by the addition of inoculated green coffee and pure fungal DNA to uncontaminated green coffee samples. The A. ochraceus DNA content of 30 naturally contaminated green coffee samples was determined and compared with the ochratoxin A concentrations.
CONCLUSIONS
Aspergillus ochraceus can be rapidly and specifically detected in green coffee by PCR. A positive correlation between the ochratoxin A content and the DNA quantity was established.
SIGNIFICANCE AND IMPACT OF THE STUDY
This work offers a quick alternative to the conventional mycological detection and quantification of A. ochraceus in green coffee.
目的
本研究旨在检测和定量生咖啡豆中产生赭曲霉毒素的真菌——赭曲霉的DNA，并将结果与天然污染样品中的赭曲霉毒素A含量进行比较。
方法与结果
测试了一种基于超声处理和商用试剂盒组合的DNA提取方案，用于回收真菌DNA。建立了用于检测赭曲霉的PCR和实时PCR方案。通过向未受污染的生咖啡豆样品中添加接种的生咖啡豆和纯真菌DNA来检查PCR的灵敏度。测定了30个天然污染的生咖啡豆样品中赭曲霉的DNA含量，并与赭曲霉毒素A浓度进行比较。
结论
通过PCR可在生咖啡豆中快速、特异性地检测出赭曲霉。建立了赭曲霉毒素A含量与DNA数量之间的正相关关系。
研究的意义和影响
这项工作为传统的生咖啡豆中赭曲霉的真菌学检测和定量提供了一种快速替代方法。
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