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

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. Graphical abstract

Advances in Laboratory Diagnosis of Coronavirus Infections in Cattle

“…The use of RT-LAMP was taken further by researchers in Germany who designed a duplex device aimed at diagnosing viral agents, particularly respiratory pathogens [65]. Their device could detect bovine coronavirus and influenza A/X-31 virus (they used this combination as a model for human respiratory infection that might include SARS-CoV-2), and the fully integrated system performed well for both pathogens [65].…”

“…The use of RT-LAMP was taken further by researchers in Germany who designed a duplex device aimed at diagnosing viral agents, particularly respiratory pathogens [65]. Their device could detect bovine coronavirus and influenza A/X-31 virus (they used this combination as a model for human respiratory infection that might include SARS-CoV-2), and the fully integrated system performed well for both pathogens [65]. The device was designed with all of the primers, reagents, and a lateral flow strip included and thus did not require additional specialized equipment, and it produced a color band to indicate a positive reaction.…”

“…The limit of detection for BCoV using purified samples was 2000 RNA copies/mL, and this was even lower for influenza A. Unfortunately, however, this was not tested with feces or clinical samples [65]. Further considerations are that the assay in this state largely reports a qualitative result (positive or negative) with no quantification, and the authors did not report on the specificity and potential cross-reactions with other similar viruses beyond the two tested.…”