Mini-Disk Capillary Array Coupling with LAMP for Visual Detection of Multiple Nucleic Acids using Genetically Modified Organism Analysis as an Example

Abstract

Convenient, portable, and low-cost multiplex nucleic acid testing (NAT) systems are the trends in the fields of food safety, environmental microorganisms, molecular diagnosis, etc. In this study, we developed a novel system for visual monitoring of multiple nucleic acids combining a mini-disk capillary array (diameter = 17 mm, embedded with 6–10 capillaries), visual loop-mediated isothermal amplification (LAMP), and quick DNA extraction called mDC-LAMP. The performance and applicability of mDC-LAMP in testing multiple nucleic acids were evaluated and verified employing genetically modified contents analysis as an example. All of the results confirmed that mDC-LAMP has the advantages of high specificity without any cross contamination, high sensitivity with a limit of detection of 25 copies/reaction, high throughput with flexible channel sensors, easy fabrication, and low costs. We believe that mDC-LAMP is a competitive choice for on-spot monitoring of multiple nucleic acids in terms of the easy fabrication/operation, low costs, and suitable performance presented in the nucleic acids test.

Isothermal nucleic acid amplification technology in HIV detection

“…However, since the LAMP technique requires multiple amplification primers for amplification reactions, multiple primers coexist when performing multiplex analysis, resulting in competitive binding of these primers to the DNA polymerase and substrate. 69 Therefore, conventional LAMP technology makes it difficult to perform multiplex testing of different genomes in the same reaction system or multiplex testing of different pathogens in the same patient. In the later period, researchers aimed at this deficiency, optimized it on the basis of detection of amplification by the release of quenching LAMP (DARQ LAMP), and realized single-tube multi-gene detection using BST 3.0, but this method has not yet been applied to HIV nucleic acid detection.…”

Advanced Technologies in On-Site Detection of Genetically Modified Products

“…Li et al designed a mini-disk capillary (Figure 3) with a detection limit of 25 copies/reaction by combining quick DNA extraction procedures with visualized LAMP findings to create the mDC-LAMP. Using transgenic maize and transgenic rice for testing, the mDC-LAMP was shown to provide the benefits of high specificity, a lack of cross-contamination, high sensitivity, high throughput, and suitability for field detection [21]. Loo et al developed a portable and rapid system for the detection of transgenic papaya using LAMP and an integrated microfluidic platform using a microfluidic lab-on-a-disc (LOAD).…”

The Potential Use of Isothermal Amplification Assays for In-Field Diagnostics of Plant Pathogens

“…In this review we have focused on paper-based lab-on-a-chip approaches that utilize simple, complex, and technological detection. Not only are pathogens detected using this approach but also genetically modified organisms [252]. Two lab-on-a-chip assays were proposed for the detection of plant pathogens.…”