Journal
FOOD ANALYTICAL METHODS
Volume 16, Issue 6, Pages 997-1006Publisher
SPRINGER
DOI: 10.1007/s12161-022-02242-1
Keywords
Recombinase Polymerase Amplification; RPA; Lateral flow; Fast molecular methods; Spoilage fungi; Internal transcribed spacer; ITS
Categories
Ask authors/readers for more resources
A rapid and reliable RPA-LF assay was developed for the detection of spoilage fungi in fruit-based products. The method showed high sensitivity and specificity, making it suitable for point of care applications.
Recombinase Polymerase Amplification combined with Lateral Flow (RPA-LF) detection was reported to be a good alternative to traditional culture-based methods, as well as other molecular techniques such as qPCR, PCR and Loop-Mediated Isothermal Amplification. The reasons for this are its simplicity, high sensitivity/specificity and ability for point of care (POC) applications. In this study, an RPA-LF assay was developed and evaluated for the detection of spoilage fungi in fruit-based products. In this sense, the universal primers ITS3 and ITS4 were labelled with digoxigenin and biotin, respectively. The method proved to be very sensitive with the ability to detect down to 1.2 pg/mu L of pure fungal DNA. Furthermore, when the assay was applied in artificially contaminated samples, low detection limits were determined, in particular 1.0 CFU/50 g and 45.7 spores/50 g for yeasts and moulds, respectively. Overall, a rapid (24-48 h) and reliable assay was developed for the detection of fungi that could be applied for POC applications.
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