Viral Detection off Swabs Using the Bead Ruptor Elite

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.