This presentation titled “Detection of Congenital Cytomegalovirus by testing Neonatal Saliva in Minutes Using a Disposable Cartridge in a Near-Patient Platform” was given by Vamsee Pamula, Founder & President of Baebies, in a livestream on August 4, 2020 hosted by the Pediatric Academic Societies 2020 PAS Summer Webinar Series – Neonatal Infectious Diseases & Immunology
Funding for this project: National Institute on Deafness And Other Communication Disorders of the NIH: R44DC016576
FINDER and associated assays are not available for sale in the US.
This presentation is based on the abstract that was accepted for an oral talk at PAS 2020:
Background: Congenital cytomegalovirus (cCMV) is the most common congenital infection and a leading cause of hearing loss and intellectual disability. In the U.S., 1 in 150 newborns are estimated to have cCMV, however, most infants have no clinically detectable symptoms and therefore, are not identified. Identification of infants with cCMV can facilitate early detection of CMV-associated hearing loss and provide interventions including antiviral therapy to improve outcomes. Although most newborn screening (NBS) is performed using dried blood spots (DBS), the sensitivity of CMV detection in DBS is a challenge, even when using DNA amplification whereas saliva and urine CMV detection have high sensitivity. Due to lack of infrastructure for transport of saliva to NBS labs, the test will likely need to be performed in hospitals similar to bilirubin, hearing screening, and pulse oximetry.
Design/Methods: CMV sequences published by Boppana and custom sequences for internal control lambda were used. Heaters and sensors were integrated into a digital microfluidic (DMF) cartridge to enable rapid amplification of target DNA sequences using fluorescent probe chemistries (R44DC016576). Reaction droplets (containing sample, master mix, primers, probes, and lambda DNA) were subjected to 50 cycles including denaturing (93°C) and extension (60°C) stages. Saliva samples from newborns are run on the cartridge.
Results: The entire 50 cycle reaction, including on-cartridge droplet dispensing, reagent mixing, and thermal cycling, was completed in 5 min. Cycle thresholds were measured from PCR curves for CMV at concentrations from 10-100,000 cp/µL with the results following expected dose response. Figure shows PCR from 1,500 CMV cp/µL multiplexed with lambda. CMV patient samples showed amplification while the normal samples did not.
Conclusion(s): This study demonstrates performance of cCMV PCR assays in a disposable cartridge in a completely automated fashion to enable in-hospital screening. All reagents are fully integrated into the cartridge eliminating the need for reagent preparation or loading. Rapid PCR (in less than 5 mins) circumvents the necessity to setup infrastructure to transport saliva samples and can accelerate adoption of near-patient testing and rapid return of results. Further testing is required to establish the performance and clinical utility of this device in a clinical setting.
AUTHORS/INSTITUTIONS: Wu, R.L. Kitchener, A. Kennedy, R. Ng, R. Sista, V. Pamula, Baebies, Inc, Durham, North Carolina; S. Permar, Duke Universiy Medical Center, Durham, North Carolina; S.B. Boppana, Pediatrics, University of Alabama at Birmingham, Birmingham, Alabama
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