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Detection and quantification of Aspergillus ochraceus in green coffee by PCR
Author(s) -
Schmidt H.,
Bannier M.,
Vogel R.F.,
Niessen L.
Publication year - 2004
Publication title -
letters in applied microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.698
H-Index - 110
eISSN - 1472-765X
pISSN - 0266-8254
DOI - 10.1111/j.1472-765x.2004.01524.x
Subject(s) - aspergillus ochraceus , ochratoxin a , aspergillus , ochratoxins , ochratoxin , green coffee , biology , food science , dna extraction , mycotoxin , polymerase chain reaction , microbiology and biotechnology , gene , biochemistry
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.