Integrated microfluidic loop-mediated-isothermal-amplification system for rapid diagnosis of aquaculture virus

The micro polymerase chain reaction (PCR) chip has been extensively reported for rapid clinical diagnosis recently. However, a series of heating and cooling processes may increase total reaction time and complexity of the control system. Besides, nucleic acid extraction, which is a critical procedure for molecular diagnosis, still remains an issue for most of the microfluidic systems. This study therefore, for the first time, reports a new automatic assay for ribonucleic acid (RNA) extraction and one-step reverse-transcription loop-mediated isothermal amplification (RT-LAMP) processes under a constant temperature for rapid detection of viruses from infected tissue samples by utilizing an integrated microfluidic system. Experimental data showed that the entire process for molecular diagnosis of an aquaculture virus from grouper tissues can be automatically performed within a short period of time and carried out with high sensitivity and specificity.