Currently, one of the most reliable detection methods for viral pathogens are polymerase chain reaction (PCR) based assays. These assays often involve procedures of swabbing a patient, processing the sample to lyse the virus, purify its nucleotides, and then process the purified genetic material via PCR for detection of a gene product needed to confirm the patient’s suspected diagnosis. This process is time-consuming and is dependent on the availability of the reagents and plastics required to complete lysis, extraction, purification, and amplification procedures. Using human coronavirus 229-E (HCoV-229E) as our model system, we have developed a method to detect virus from an in vitro spiked swab using only mechanical lysis via the Omni Bead Ruptor Elite bead mill homogenizer and a direct-to-PCR methodology, bypassing the reagent-heavy and time-consuming extraction and purification steps.
Figure 1: HCoV-229E N-Gene amplification via RT-qPCR following homogenization of virally-spiked swabs. Red, 1.2e6 PFU/mL spiked swab. Brown, 1.2e5 PFU/mL spiked swab. Pink, 1.2e4 PFU/mL spiked swab. Navy, 1.2e3 PFU/mL spiked swab. Teal, 1.2e2 PFU/mL spiked swab. Olive, 1.2e1 PFU/mL spiked swab. Black, 1.2e0 PFU/mL spiked swab. Green, negative control.