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. 2023 Sep 8;25(4):36.
doi: 10.1007/s10544-023-00676-w.

A fully integrated duplex RT-LAMP device for the detection of viral infections

Nicolas Mytzka  1 , Skaiste Arbaciauskaite  2   3 , Natalia Sandetskaya  1 , Kai Mattern  1 , Dirk Kuhlmeier  1
Affiliations

A fully integrated duplex RT-LAMP device for the detection of viral infections

Nicolas Mytzka et al. Biomed Microdevices. .

Abstract

Respiratory viruses can cause epidemics or pandemics, which are worldwide outbreaks of disease. The severity of these events varies depending on the virus, its characteristics, along with environmental factors. The frequency of epidemics and pandemics caused by respiratory viruses is difficult to predict, but the potential severity of such events underlines the importance of continued monitoring, research, and preparation for emerging infectious diseases. To help improve pandemic preparedness, we created a fully integrated duplex reverse transcription loop-mediated isothermal amplification (RT-LAMP) device targeting two respiratory viruses, influenza A/X-31 virus and bovine coronavirus, as a replacement for SARS-CoV-2. This device can be adapted to any other respiratory virus. In this study, we showed and evaluated a prototype of a microfluidic system, and showed that duplex RT-LAMP can detect and distinguish between the two viruses, with LoDs of 2,000 copies/ml for bovine coronavirus and 200 copies/ml for influenza A/X-31 virus.

Keywords: Coronavirus; Influenza; Multiplex; Point-of-care; RT-LAMP; Respiratory infections; SARS-CoV-2.

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Conflict of interest statement

The authors declare no conflict of interest.

Figures

Fig.1
Fig.1
Microfluidic chip design of the duplex RT-LAMP device. Chip design with microfluidic channels, connecting the reaction chamber that contains lyophilized LAMP reagents (colored with phenol red) with the inlet, the integrated LFT and the water-filled blister
Fig. 2
Fig. 2
Duplex RT-LAMP assay for influenza A virus and bovine coronavirus. LFT bands from left to right: control line (c), InfA test line (i), BovCov test line (b); A Negative control with water; B Positive control with 1,000 RNA copies of InfA and BovCov
Fig. 3
Fig. 3
Quantitative RT-LAMP for bovine coronavirus and influenza A virus. RT-LAMP amplification curves for A bovine coronavirus and B influenza A RNA dilution series detected with LightCycler
Fig. 4
Fig. 4
LoD for RT-LAMP device for bovine coronavirus and influenza A virus. A Three positive replicates for 100 RNA copies of BovCov and 10 RNA copies of InfA representing the LoD of the respective LAMP reaction; B Two positive replicates (top, bottom) and one negative replicate (middle) for BovCov LAMP, two weak positive replicates (top, bottom) and one negative replicate (middle) for InfA LAMP
See this image and copyright information in PMC

References

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Supplementary concepts