SBIR-STTR Award

?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 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 in 2 to 3 days which often does not provide timely results for neonatal sepsis cases. Typical neonatal patients are weighing 1,000 grams or less and their total blood volume may be as low as ~ 50 mL, thus the iatrogenic blood loss for premature babies may represent up to 12% - 31% of total blood loss due to phlebotomy. New point-of-care methods with faster detection time and using smaller amounts of sample are urgently needed. Molecular assays based on DNA specificity for sepsis pathogens represent a promising diagnostic tool for early identification of bacteremia. However, today 90% of hospitals in USA 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 (maximum 4-6) 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 min and that is capable of using very small sample volumes, as low as 2 - 200 microliters, compatible with the neonatal sepsis detection. Multiplexed identification of the neonatal sepsis pathogens is enable using the microarray approach. In addition, a number of innovations are offered in the project that will bring a 5 fold decrease in the cost of instrumentation and cartridges compared to the commercialized platforms. The platform also offers a unique new qualitative parameter in DNA/RNA diagnostics that will enable integration of data for both the length of the nucleic acids as well as it will preserve the strand specificity, e.g., important in alternative splicing mechanisms. The assay development in the Phase I will focus on representative neonatal sepsis pathogens and will establish the basic platform, and cartridge prototypes and will provide a thorough validation of the assays developed using de- identified archived clinical samples.

Public Health Relevance Statement:


Public Health Relevance:
Neonatal sepsis and/or bacteremia is a life-threatening medical emergency for infants in the Rapid diagnosis of sepsis in NICUs (Neonatal Intensive Care Units) and cause over one million deaths per year worldwide. Another problem is the large volume of blood withdrawn for diagnosis critically affecting neonates. New point- of-care methods with faster detection time and using smaller amounts of sample are urgently needed. This project introduces a highly innovative point-of-care (POC) method for diagnostics of neonatal sepsis that will approach total assay turn-around time of ~15 min and that is capable of using very small sample volumes, as low as 2 - 200 microliters, compatible with the neonatal sepsis detection. The platform will find broad applications in the point-of-care diagnostics of emergent, pandemic and infectious diseases.

NIH Spending Category:
Bioengineering; Genetics; Hematology; Infant Mortality; Infant Mortality/ (LBW); Infectious Diseases; Patient Safety; Pediatric; Perinatal - Birth - Preterm (LBW); Perinatal Period - Conditions Originating in Perinatal Period; Septicemia

Project Terms:
Affect; Alternative Splicing; Antibiotics; Archives; assay development; Bacteremia; Bacteria; base; Biological Assay; Blood; Blood Circulation; Blood specimen; Blood Volume; Carbon; Cause of Death; Cessation of life; Chemistry; Clinical; Collaborations; Color; Communicable Diseases; cost; Country; data integration; design; Detection; Development; Device or Instrument Development; Diagnosis; Diagnostic; diagnostic assay; Diagnostic Procedure; DNA; DNA amplification; DNA analysis; Drug resistance; drug resistant microorganism; Early identification; Electronics; Electrophoresis; Escherichia coli; Fluorescence; fungus; gastrointestinal; Genus staphylococcus; Goals; Gold; Haemophilus influenzae; Hemorrhage; Hospitals; Infant; Ink; innovation; instrument; instrumentation; Lead; Length; Life; Listeria monocytogenes; Low income; Marketing; Medical emergency; medical schools; Methods; Miniaturization; Modeling; Molecular; Neonatal; Neonatal Intensive Care Units; neonatal sepsis; neonate; next generation; nucleic acid detection; Nucleic Acids; Optics; Organism; pandemic disease; pathogen; pediatric department; Phase; point of care; point-of-care diagnostics; Premature Infant; Preparation; Price; prototype; public health relevance; rapid diagnosis; rapid technique; Reaction Time; Risk; RNA; RNA analysis; Sampling; Sensitivity and Specificity; Sepsis; Specificity; Streptococcus; Streptococcus Group B; Testing; Time; time use; tool; Validation; Venous blood sampling; Weighing patient

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.

Public Health Relevance Statement:
Project Narrative 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.

Project Terms:
Accident and Emergency department; accurate diagnosis; Acquired Immunodeficiency Syndrome; Antibiotic Resistance; Antibiotic Therapy; Antibiotics; Archives; assay development; Bacteremia; Bacteria; Bacterial Antibiotic Resistance; base; Biological Assay; Birth; Blood; Blood Circulation; Blood specimen; Blood Volume; Carbon; Centers for Disease Control and Prevention (U.S.); Cessation of life; Child; Childhood; Clinic; Clinical; Color; Communicable Diseases; cost; Data; Decentralization; design; Detection; Development; Diagnosis; Diagnostic; diagnostic accuracy; DNA; DNA analysis; Drug resistance; drug resistant microorganism; Early identification; early onset; Extremely Low Birth Weight Infant; Fatality rate; fungus; gastrointestinal; Genes; Goals; Gold; Guidelines; Hemorrhage; Hospitals; Hour; Iatrogenesis; improved; Incidence; Infant; Infection; innovation; instrument; instrumentation; Lead; Life; Low income; low income country; malignant breast neoplasm; Malignant neoplasm of prostate; Medical emergency; Methods; microorganism; Molecular; Monitor; Mothers; multiplex detection; National Institute of Child Health and Human Development; Neonatal; Neonatal Intensive Care Units; neonatal sepsis; neonate; novel diagnostics; Organism; pandemic disease; pathogen; Pathogen detection; Patients; Phase; point of care; point-of-care diagnostics; Premature Infant; prevent; Price; rapid diagnosis; rapid technique; Reaction Time; Regimen; Risk; RNA; Sampling; Sensitivity and Specificity; Sepsis; Specificity; ST14 gene; System; Technology; Testing; Time; time use; tool; urgent care; Urine; Validation; Vendor; Very Low Birth Weight Infant; Whole Blood