Severe sepsis strikes more than a million patients in USA per year and it has been estimated that between 28% and 50% percent of these patients die, exceeding the number of U.S. deaths from prostate cancer, breast cancer and AIDS combined. Neonatal sepsis causes over one million death per year worldwide and 5% to 60% of infants treated with antibiotics die despite the treatment, with higher rates occurring in low income countries. Rapid and accurate diagnosis of sepsis in NICUs (Neonatal Intensive Care Units) is crucial for implementing a timely treatment. Because sepsis is a life-threatening medical emergency a large proportion of infants receive treatment with potent systemic antibiotics that can adversely lead to destruction of the infant?s normal gastrointestinal flora and the risk of becoming colonized with drug-resistant microorganisms. Blood culturing is used as the gold standard for neonatal diagnosis of sepsis, however the samples are returned after 48 hrs which often does not provide timely results for neonatal sepsis treatment. Multiple sampling that will increase accuracy of neonatal diagnosis by culturing is minimized because the iatrogenic blood loss may represent up to 12% - 31% of total blood in premature babies. New point-of-care methods with faster detection time and more accurate to use smaller amounts of sample are urgently needed. Molecular DNA-based assays have been introduced into the CDC Guidelines for neonatal sepsis diagnosis but current methods lack accuracy, need enrichment of samples and cannot provide the needed multiplexed pathogen identification. However, today 90% of hospitals in USA and world cannot perform DNA analysis and samples are sent to the centralized reference labs with the response time of 2-3 days. The current point-of-care diagnostics are mostly based on the real-time PCR methods that are limited by the number of colors for detecting multiple organisms or by the high price of the instrument or assay cost. This project introduces a new point-of-care (POC) rapid method for DNA/RNA diagnostics that will approach total assay turn-around time of ~15-20 min and that can use very small sample volumes, as low as 5 - 200 microliters, compatible with the neonatal sepsis detection. Multiplexed identification of the neonatal sepsis pathogens is enabled using the microarray approach. The proposed technology will enable a new diagnostic approach that could more rapidly identify neonatal pathogens including hospital associated infections and antibiotic resistant pathogens yielding a more tailored administration of regimen of antibiotics and promoting a judicious use of antibiotics. Several innovations in manufacturing of cartridge disposable and instrument will bring a 5-8-fold decrease in the cost of instrumentation and cartridges compared to the commercialized platforms. The Phase II project will focus on the platform and neonatal sepsis/HAI assay development; however, it is envisioned that the platform will find other broad applications in decentralized, urgent care settings as well as in hospitals.
Neonatal sepsis is a life-threatening medical emergency for infants in the NICUs (Neonatal Intensive Care Units) and causes over one million deathS per year worldwide. Standard culturing method for diagnosis of sepsis requires up to 48 hrs that is often too late for implementing an adequate antibiotic treatment, especially when trying to minimize unnecessary antibiotic treatments and/or impact the development of antibiotic resistant bacterial strains. Standard methods require a relatively large blood sample volumes causing significant iatrogenic blood losses in neonates, that limit the repetition of the sampling, that would improve the accuracy and prevent false positives due to bacterial contamination during sampling. This project brings a new DNA-based point-of-care method with ~15-20 minutes sample-to-answer time, offering very small sample volumes, as low as 5 - 200 microliters, compatible with the neonatal sepsis detection and a highly-multiplexed detection of both sepsis and antibiotic resistant pathogens in neonatal infections. The platform will find broad applications in the point-of-care diagnostics of emergent, pandemic and infectious diseases as well as its implementation in decentralized settings such as urgent care clinics and emergency rooms as well as in hospitals.
