TY - JOUR
T1 - Development and application of a loop-mediated isothermal amplification assay for rapid identification of aflatoxigenic molds and their detection in food samples
AU - Luo, Jie
AU - Vogel, Rudi F.
AU - Niessen, Ludwig
N1 - Funding Information:
JL was supported by the China Scholarship Council, China State-Sponsored Postgraduate Study Abroad Program . We thank Prof. Jens C. Frisvad for providing pure cultures of aflatoxin producing fungi and S. Dummert for technical assistance.
PY - 2012/10/15
Y1 - 2012/10/15
N2 - Aflatoxins are the most thoroughly studied mycotoxins. They are produced by several members of the genus Aspergillus in section Flavi with Aspergillus flavus, Aspergillus parasiticus, and Aspergillus nomius being frequently isolated from contaminated food sources. In this work, we describe the development and evaluation of loop-mediated isothermal amplification (LAMP) assays for rapid detection of the three species in separate analyses. The acl1-gene of A. flavus and amy1-genes of A. nomius and A. parasiticus were used as target genes. The detection limits were 2.4, 7.6 and 20. pg of pure DNA/reaction for A. flavus, A. nomius and A. parasiticus, respectively. For specificity testing, DNA extracted from mycelia of representative strains of 39 Aspergillus species, 23 Penicillium species, 75 Fusarium species and 37 other fungal species was used as a template for the specific LAMP primer sets developed for the three target species. The LAMP assay was combined with a DNA extraction method for the analysis of pure fungal cultures as well as artificially contaminated Brazil nuts, peanuts and green coffee beans. It is suggested that the developed LAMP assay is a promising tool in the prediction of a potential aflatoxin risk in food and food raw materials and may therefore be suitable for high throughput analysis in the food industry.
AB - Aflatoxins are the most thoroughly studied mycotoxins. They are produced by several members of the genus Aspergillus in section Flavi with Aspergillus flavus, Aspergillus parasiticus, and Aspergillus nomius being frequently isolated from contaminated food sources. In this work, we describe the development and evaluation of loop-mediated isothermal amplification (LAMP) assays for rapid detection of the three species in separate analyses. The acl1-gene of A. flavus and amy1-genes of A. nomius and A. parasiticus were used as target genes. The detection limits were 2.4, 7.6 and 20. pg of pure DNA/reaction for A. flavus, A. nomius and A. parasiticus, respectively. For specificity testing, DNA extracted from mycelia of representative strains of 39 Aspergillus species, 23 Penicillium species, 75 Fusarium species and 37 other fungal species was used as a template for the specific LAMP primer sets developed for the three target species. The LAMP assay was combined with a DNA extraction method for the analysis of pure fungal cultures as well as artificially contaminated Brazil nuts, peanuts and green coffee beans. It is suggested that the developed LAMP assay is a promising tool in the prediction of a potential aflatoxin risk in food and food raw materials and may therefore be suitable for high throughput analysis in the food industry.
KW - Aflatoxin
KW - Aspergillus
KW - Detection
KW - Food
KW - Loop-mediated isothermal amplification (LAMP)
UR - http://www.scopus.com/inward/record.url?scp=84868033493&partnerID=8YFLogxK
U2 - 10.1016/j.ijfoodmicro.2012.08.018
DO - 10.1016/j.ijfoodmicro.2012.08.018
M3 - Article
C2 - 23107500
AN - SCOPUS:84868033493
SN - 0168-1605
VL - 159
SP - 214
EP - 224
JO - International Journal of Food Microbiology
JF - International Journal of Food Microbiology
IS - 3
ER -