Non-invasive, ultra-sensitive, high-throughput assays to quantify rare biomarkers in the blood.
Many diseases are easier to treat and control when detected at an early stage of disease progression. Often, disease-related antigens or biomarkers are shed from the primary site and present in the blood. Unfortunately, there are very few tests capable of detecting these rare biomarkers in the blood. A blood test would be very useful to diagnose the disease earlier, monitor effectiveness of treatments, predict recurrence, and monitor recurrence. There is certainly a need to develop assays that are ultra-sensitive, non-invasive, and high-throughput. Here we describe several highly sensitive immunological assays we have developed to detect rare serum antigens. Initially we created an assay named immuno-detection amplified by T7 RNA polymerase (IDAT). To enhance the effectiveness and streamline the procedure, this assay was amended to the facile amplification system termed fluorescent amplification catalyzed by T7 polymerase technique (FACTT). These assays have been used to analyze the tumor antigen HER2 and the prion protein PrPSc. They can also be applied to other tumor markers or antigens from a variety of diseases such as cardiovascular disease, rheumatoid arthritis, Alzheimer's disease, Parkinson's disease, and hepatitis. These tests are not limited to testing only serum, but may also be applicable to detecting biomarkers in tissue, saliva, urine, cerebrospinal fluid, etc. Clearly, the FACTT-based technology represents an important step in the detection of rare molecules in fluids or tissues for a variety of diseases.
Department of Pathology & Laboratory Medicine, University of Pennsylvania, 252 John Morgan Building, 3620 Hamilton Walk, Philadelphia, PA 19104-6082, USA.
Jaclyn A
JA
Freudenberg
Kathryn
K
Bembas
Mark I
MI
Greene
Hongtao
H
Zhang
D000818:Animals; D000941:Antigens / Q000097:blood; D001681:Biological Assay / Q000639:trends; D015415:Biomarkers / Q000097:blood* / Q000134:cerebrospinal fluid / Q000652:urine; D012321:DNA-Directed RNA Polymerases; D019937:Diagnostic Techniques and Procedures / Q000639:trends*; D005456:Fluorescent Dyes; D006801:Humans; D007124:Immunoenzyme Techniques / Q000639:trends*; D007364:Intercalating Agents; D016133:Polymerase Chain Reaction / Q000379:methods; D018620:PrPSc Proteins / Q000032:analysis; D016601:RNA-Binding Proteins / Q000032:analysis; D018719:Erb-b2 Receptor Tyrosine Kinases / Q000032:analysis; D014764:Viral Proteins
D000941:Antigens; D015415:Biomarkers; D012321:DNA-Directed RNA Polymerases; D005456:Fluorescent Dyes; D007364:Intercalating Agents; D018620:PrPSc Proteins; D016601:RNA-Binding Proteins; D018719:Erb-b2 Receptor Tyrosine Kinases; D014764:Viral Proteins; C062210:bacteriophage T7 RNA polymerase