Influenza viruses are highly contagious negative-sense RNA viruses cause approximately 114,000 hospitalizations and 20,000 deaths annually. Several drugs are approved by the US Food and Drug Administration (FDA) for treating influenza infections, but successful treatment depends on knowing the specific antigenic subtype and administering these drugs within the first 48 h of illness. Recent outbreaks of avian influenza H5N1 in East Asia have fueled concern over another pandemic and emphasize the need for comprehensive strain sub-typing in surveillance and diagnostic applications. The objective of this Phase 2 Project is therefore to develop and field test a sample-to-answer RT-PCR array kit for influenza detection and antigenic sub-typing in diagnostic and surveillance settings, with an emphasis on simple, low-cost microarray technology that can be deployed and used in peripheral hospital labs. The performance goal is a sample-to answer result in 2 hours or less at an input target concentration of 102 viral particles per mL nasopharyngeal swab. Early adopters at Wadsworth Center, Columbia University, Little Company of Mary Hospital and the CDC will assist in the pre-clinical verification of the technology on amended and blind clinical samples.
The influenza RT-PCR array will be the first-of-its-kind combination of PCR and microarray technology in a simple, inexpensive test kit for disease surveillance and diagnosis. The underlying platform developed herein will likewise find broad application in many areas of infectious disease diagnostics.
|Chandler, Darrell P; Griesemer, Sara B; Knickerbocker, Christopher et al. (2014) Development and clinical testing of a simple, low-density gel element array for influenza identification, subtyping, and H275Y detection. J Virol Methods 208:152-9|
|Holmberg, Rebecca C; Gindlesperger, Alissa; Stokes, Tinsley et al. (2013) High-throughput, automated extraction of DNA and RNA from clinical samples using TruTip technology on common liquid handling robots. J Vis Exp :e50356|
|Griesemer, Sara B; Holmberg, Rebecca; Cooney, Christopher G et al. (2013) Automated, simple, and efficient influenza RNA extraction from clinical respiratory swabs using TruTip and epMotion. J Clin Virol 58:138-43|
|Chandler, Darrell P; Bryant, Lexi; Griesemer, Sara B et al. (2012) Integrated Amplification Microarrays for Infectious Disease Diagnostics. Microarrays (Basel) 1:107-24|
|Chandler, Darrell P; Griesemer, Sara B; Cooney, Christopher G et al. (2012) Rapid, simple influenza RNA extraction from nasopharyngeal samples. J Virol Methods 183:8-13|