Color-Coding of Microchip RT-PCR Test System for SARS-CoV-2 Detection

An RT-PCR based microchip test system for the detection of SARS-CoV-2 offers pre-loaded and lyophilized reagents in microchip. However, 30- 48-microwell formats being miniaturized performing 1.2 μl reaction, seek visual attention during sample addition. Therefore, adding colorants as color indicator matrix microchips or to master mix can impart not only user-friendliness task liquid handling but also precision, color-codes easy identification multiple kits layout without compromising PCR data quality. A panel was screened their background intensity, spectral inertness towards channels AriaDNATM analyzer, interference with reporter dyes (FAM, Cy5 ROX), visibility optimal concentration microwell. The colorant displaying insignificant impact on quality amplification (Ct, fluorescence, sensitivity) comparison no-colorant control chosen inclusion kit. Tartrazine, Acid Red, Brilliant Blue FAST Green were found be stable suitable. Storage Fast tested at 40°C, 22°C, 4°C, -20°C 70 days suitable compatible different mixes available lyophilized. Additionally, presence absence two Tartrazine validated clinical samples SARS-COV-2. No significant these both intra- inter-microchips observed Ct intensity control. suggested that used sample, user-friendly empty microchips. For reagents, added mixture precision color-coding manufacturing improve loaded reagent mix.