Paper-based colorimetric loop-mediated isothermal amplification (LAMP) assay for the identification of latent Colletotrichum in harvested fruit

Manpreet Kaur; Khadijah Ayarnah; Danielle Duanis-Assaf; Noam Alkan; Evgeni Eltzov

doi:10.1016/j.aca.2023.341394

Paper-based colorimetric loop-mediated isothermal amplification (LAMP) assay for the identification of latent Colletotrichum in harvested fruit

Anal Chim Acta. 2023 Aug 1:1267:341394. doi: 10.1016/j.aca.2023.341394. Epub 2023 May 18.

Authors

Manpreet Kaur¹, Khadijah Ayarnah¹, Danielle Duanis-Assaf¹, Noam Alkan², Evgeni Eltzov³

Affiliations

¹ Department of Postharvest Science, Institute of Postharvest and Food Sciences, The Volcani Institute, Agricultural Research Organization, Bet Dagan, 50250, Israel; Institute of Biochemistry, Food Science and Nutrition, Faculty of Agriculture, Food and Environment, The Hebrew University of Jerusalem, Rehovot, 76100, Israel.
² Department of Postharvest Science, Institute of Postharvest and Food Sciences, The Volcani Institute, Agricultural Research Organization, Bet Dagan, 50250, Israel.
³ Department of Postharvest Science, Institute of Postharvest and Food Sciences, The Volcani Institute, Agricultural Research Organization, Bet Dagan, 50250, Israel; Agro-Nanotechnology and Advanced Materials Research Center, Institute of Postharvest and Food Science, Department of Postharvest Science, Agricultural Research Organization, The Volcani Institute, Rishon LeZion, 7505101, Israel. Electronic address: eltzov@volcani.agri.gov.il.

PMID: 37257967
DOI: 10.1016/j.aca.2023.341394

Abstract

Paper-based analytical devices (PADs) have gained enormous attention because of their low-cost, simple fabrication, and portability. Here, we propose a paper-based device for performing reverse transcription loop-mediated isothermal amplification (RT-LAMP) with real-time simultaneous detection of C. gloeosporioides latent infections in tomatoes. RT-LAMP-based PAD platform comprises a paper substrate on which the DNA amplification reaction occurs. Among different types of tested papers, cellulose membrane (grade 4) enabled effective visualization of the amplification result. The assay was found highly selective for the latent stage of C. gloeosporioides with lower limit of detection (LOD) of 0.5 pg of total extracted RNA. The developed assay generated the results within 40 min and hence can be efficiently employed for identifying C. gloeosporioides in resource-limited settings.

Keywords: Fruit; Fungal pathogen; Loop-mediated isothermal amplification (LAMP); Paper-based analytic devices (PADs); Point of care technology (POCT); mRNA.

MeSH terms

Colletotrichum* / genetics
Colorimetry / methods
Fruit
Molecular Diagnostic Techniques
Nucleic Acid Amplification Techniques / methods
Reverse Transcription
Sensitivity and Specificity

Supplementary concepts

LAMP assay