Limit of Detection (LOD) tested on inactivated SARS-CoV-2 viral particles at 250, 500, 1000, 2000 and 4000 cp/ml. Type of sample diluted in UTM + cell line KG1A at different cp/reaction. All samples tested with KG1A (cell line) amount into the reaction: 1200 cells per reaction. LOD of 8 copies/reaction offers the test sensitivity required.
The kit is compatible with samples collected in non-fixation transport media like UTM, VTM, PBS, 0.9% NaCl, Virocult and eSwab. 8 µl of respective transport medium spiked with indicated amount of IVT + 2 µl Viral RNA UM Prep Buffer + 10 µl Viral RNA UM Master Mix show comparable Ct values.
An aliquot is taken from a primary sample (nasopharyngeal, oropharyngeal or nasal swab) in transport media as starting material. This is added to an optimized sample preparation buffer, which allows preparation of the viral RNA template without degradation in two minutes, before being combined with an RT-qPCR reaction mix for rapid amplification on any thermocycler using any assay. The output is finally interpreted, delivering test results in under one hour.
Positive high viral titer swab samples spiked into negative saliva and gargle samples. Samples heat-treated at 95°C for 15 min using the QIAprep& workflow for saliva or treated with QIAprep& Buffer AB and incubated at 95°C for 5 min.
Saliva samples (n=20). Samples 1–17: Positive donors. Samples 18–20: Negative donors. Same samples differentially treated. Protocol 1:QIAprep& Viral RNA UM Kit without Buffer AB (95°C, 15 min). Protocol 2:QIAprep& Viral RNA UM Kit with Buffer AB.