Journal
INTERNATIONAL JOURNAL OF FOOD MICROBIOLOGY
Volume 149, Issue 3, Pages 226-235Publisher
ELSEVIER SCIENCE BV
DOI: 10.1016/j.ijfoodmicro.2011.06.019
Keywords
Ochratoxin A; Mold; qPCR; Food; SYBR Green; TaqMan probes
Categories
Funding
- Spanish Comision Interministerial de Ciencia y Tecnologia [AGL2007-64639, BES-2008-008021]
- Carnisenusa [CSD2007-00016]
- Consolider Ingenio
- Junta de Extremadura [GRU08100, GRU09158]
- FEDER
- Danish Food Industry Agency [3412-07-01873]
Ask authors/readers for more resources
Ochratoxin A (OTA) is a mycotoxin synthesized by a variety of different fungi, most of them from the genera Penicillium and Aspergillus. Early detection and quantification of OTA producing species is crucial to improve food safety. In the present work, two protocols of real-time qPCR based on SYBR Green and TaqMan were developed, and their sensitivity and specificity were evaluated. Primers and probes were designed from the non-ribosomal peptide synthetase (otanpsPN) gene involved in OTA biosynthesis. Seventy five mold strains representing OTA producers and non-producers of different species, usually reported in food products, were used as references. All strains were tested for OTA production by mycellar electrokinetic capillary electrophoresis (MECE) and high-pressure liquid chromatography-mass spectrometry (HPLC-MS). The ability of the optimized qPCR protocols to quantify OTA-producing molds was evaluated in different artificially inoculated foods. A good linear correlation was obtained over the range 1 x 10(4) to 10 conidia/g per reaction for all qPCR assays in the different food matrices (cooked and cured products and fruits). The detection limit in all inoculated foods ranged between 1 and 10 conidia/g for SYBR Green assay and TaqMan. No significant differences were found between the Ct values obtained from pure mold DNA and pure mold DNA mixed with food DNA. The ability of the designed qPCR methods to quantify two known conidial suspensions inoculated on several foods was evaluated. The amount of conidia assessed by both qPCR methods was close to the inoculated amount for most foods and indicates that the described procedure holds potential for use for the detection and quantification of OTA producing molds in foods. (C) 2011 Elsevier B.V. All rights reserved.
Authors
I am an author on this paper
Click your name to claim this paper and add it to your profile.
Reviews
Recommended
No Data Available