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. 2019 Apr 10;9(1):5846.
doi: 10.1038/s41598-019-42275-x.

Loop-mediated isothermal amplification (LAMP) approach for detection of heat-resistant Talaromyces flavus species

Jacek Panek  1 , Magdalena Frąc  2
Affiliations

Loop-mediated isothermal amplification (LAMP) approach for detection of heat-resistant Talaromyces flavus species

Jacek Panek et al. Sci Rep. .

Abstract

Talaromyces flavus is a soilborne fungus that can contaminate fruits. It constitutes serious influence on heat-processed food spoilage, as T. flavus belongs to the heat-resistant fungi group, which are able to survive the pasteurization process. Moreover T. flavus has been reported to be capable of mycotoxigenicity, therefore they have a serious threat to human health. To maintain the safety of food production, sensitive method for T. flavus detection was developed. The loop mediated amplification, abbreviated LAMP, reactions were designed as specific for detection of DNA replication licensing factor gene of T. flavus. The specificity of assay was confirmed by use of 5 T. flavus strains and 35 other fungal isolates. The achieved limit of detection was 1fg of T. flavus genomic DNA and 64 ascospores in 1 g of strawberry fruits or soil samples.

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Conflict of interest statement

The authors declare no competing interests.

Figures

Figure 1
Figure 1
Positioning and orientation of regions used to design LAMP primers within the nucleotide sequence of the DNA replication licensing factor gene of T. flavus DSM 63536 (GenBank accession no. KX587558).
Figure 2
Figure 2
The specificity of Talaromyces flavus LAMP detection method. (a) The amplification plots (∆R) of T. flavus and 35 other studied isolates (Table 2) in time. Legend describes tested T. flavus isolates, while rest of tested strains are described in Table 2. NTC stands for non-template control. (b) Melting curve analysis of obtained products. DNA isolated with EURx Plant&Fungi DNA Purification Kit.
Figure 3
Figure 3
Sensitivity study of Talaromyces flavus LAMP detection method. (a) The amplification plots (∆R) of pure genomic T. flavus DSM 63536 DNA added into reaction in different concentrations in time. (b) The amplification plot (∆R) of T. flavus detected in strawberries contaminated by different number of ascospores added to 1 g of sample (a/g) prior the DNA isolation reaction. DNA isolated with EURx Plant&Fungi DNA Purification Kit. (c) The amplification plot (∆R) of T. flavus detected in soil contaminated by different number of ascospores added to 1 g of sample prior the DNA isolation reaction. NTC stands for non-template control. DNA isolated with EURx Soil DNA Purification Kit.
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References

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