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Rapid detection of aflatoxigenic Aspergillus sp. in herbal specimens by a simple, bendable, paper-based lab-on-a-chip
Piyasak Chaumpluk1, Pattra Plubcharoensook2, Sehanat Prasongsuk3
1Laboratory of Plant Transgenic Technology and Biosensor, Department of Botany, Faculty of Science, Chulalongkorn University, Bangkok, Thailand. piyasakcha@yahoo.com.
Abstract:
Postharvest herbal product contamination with mycotoxins and mycotoxin-producing fungi represents a potentially carcinogenic hazard. Aspergillus flavus is a major cause of this issue. Available mold detection methods are PCR-based and rely heavily on laboratories; thus, they are unsuitable for on-site monitoring. In this study, a bendable, paper-based lab-on-a-chip platform was developed to rapidly detect toxigenic Aspergillus spp. DNA. The 3.0-4.0 cm(2) chip is fabricated using Whatman™ filter paper, fishing line and a simple plastic lamination process and has nucleic acid amplification and signal detection components. The Aspergillus assay specifically amplifies the aflatoxin biosynthesis gene, aflR, using loop-mediated isothermal amplification (LAMP); hybridization between target DNA and probes on blue silvernanoplates (AgNPls) yields colorimetric results. Positive results are indicated by the detection pad appearing blue due to dispersed blue AgNPls; negative results are indicated by the detection pad appearing colorless or pale yellow due to probe/target DNA hybridization and AgNPls aggregation. Assay completion requires less than 40 min, has a limit of detection (LOD) of 100 aflR copies, and has high specificity (94.47%)and sensitivity (100%). Contamination was identified in 14 of 32 herbal samples tested (43.75%). This work demonstrates the fabrication of a simple, low-cost, paper-based lab-on-a-chip platform suitable for rapid-detection applications.
Insights
A new paper-based device rapidly detects dangerous Aspergillus fungi in herbal products. This low-cost, on-site test identifies contamination, crucial for food safety and preventing carcinogenic risks.
Area of Science:
- Food Science
- Analytical Chemistry
- Biotechnology
Background:
- Postharvest herbal products can be contaminated by mycotoxins and fungi, posing carcinogenic risks.
- Aspergillus flavus is a primary fungal contaminant, and current laboratory-based detection methods are not suitable for rapid, on-site monitoring.
Purpose of the Study:
- To develop a rapid, on-site detection method for toxigenic Aspergillus species DNA in herbal products.
- To create a low-cost, paper-based lab-on-a-chip platform for detecting the aflatoxin biosynthesis gene, aflR.
Main Methods:
- Fabrication of a bendable, paper-based lab-on-a-chip using filter paper, fishing line, and lamination.
- Utilized loop-mediated isothermal amplification (LAMP) to amplify the aflR gene.
- Employed blue silvernanoplates (AgNPls) for colorimetric detection of DNA hybridization.
Main Results:
- The assay detected as few as 100 aflR gene copies with high specificity (94.47%) and sensitivity (100%).
- Assay completion time was under 40 minutes, yielding visible colorimetric results.
- Contamination by toxigenic Aspergillus was identified in 43.75% of tested herbal samples (14 out of 32).
Conclusions:
- A simple, low-cost, paper-based lab-on-a-chip platform was successfully developed for rapid detection of Aspergillus DNA.
- This technology is suitable for on-site monitoring of herbal products, addressing limitations of traditional laboratory methods.
- The developed platform offers a practical solution for ensuring the safety of herbal products by detecting potential carcinogenic contaminants.
