The invention features novel attenuated dengue virus mutants and compositions thereof. The invention further features an attenuated dengue virus comprising a proline to leucine change at amino acid position 2343 of non-structural protein 4B (NS4B), wherein the numbering is based upon the prototypic isolate DEN4 Dominica 1981 and the attenuated mutant DEN virus has at least one of the following properties: improved replication in Vero cells or restricted replication in mosquito cells.

A group of mosquito-borne flaviviruses that cause fatal encephalitis in humans is among the most important of all emerging human pathogens of global significance. This group includes Japanese encephalitis virus (JEV), West Nile virus, St. Louis encephalitis virus, and Murray Valley encephalitis virus. In the present disclosure, the first reverse genetics system has been developed for SA14-14-2, a live JE vaccine that is most commonly used in most JE-endemic areas, by constructing an infectious bacterial artificial chromosome that contains the full-length SA14-14-2 cDNA. Using this infectious SA 14-14-2 cDNA, combined with a mouse model for JEV infection, a key viral neurovirulence factor has been discovered that is a conserved single amino acid in the ij hairpin adjacent to the fusion loop of the viral E glycoprotein, which regulates viral infectivity into neurons within the central nervous system.

The present invention provides kits and assay methods for the early detection of pathogens, precise identification of the etiologic agent, and improved disease surveillance. More specifically, the present invention discloses an immunoassay leading to the rapid and simultaneous detection of antibodies to a wide range of infectious pathogens in biological fluids of infected patients. This immunoassay involves the covalent and oriented coupling of fusion proteins comprising an AGT enzyme and a viral antigen on an identifiable solid support (e.g. fluorescent microspheres), said support being previously coated with an AGT substrate. This coupling is mediated by the irreversible reaction of the AGT enzyme on its substrate. The thus obtained antigen-coupled microspheres show enhanced capture of specific antibodies as compared to antigen-coupled microspheres produced by standard amine coupling procedures. The methods of the invention possess the ability to multiplex, minimize the amount of biological sample, and have enhanced sensitivity and specificity toward target antibodies as compared with classical ELISA or Radio-Immunoprecipitation assays.

Methods and devices for molecular imprinting include a molecular imprinting synthesis and matching a physiological pH of a template utilized in the molecular imprinting synthesis to achieve molecular imprinting. Molecular imprinting can be achieved by matching the physiological pH of the template used in a molecular imprinting synthesis. Furthermore, electrostatic charges can be complementary matched to the template, by obtaining crystallographic data of a protein template. Particularly, positively and negatively charged amino acids can be counted and matched by an oppositely charged monomer. For hydrophobic amino acids, isoleucin, leucin, and valine amino acids are counted. Since not all hydrophobic amino acids are exposed, the hydrophobic amino acid and hydrophobic monomer ratio can be determined experimentally by varying ratios from 1:1 to 1:10.

The present invention provides compositions and methods that can be used to determine a peptide signature for an antibody repertoire in a sample comprising multiple antibodies. The method can be used to characterize a phenotype in a sample, such as providing a diagnosis, prognosis or theranosis of a medical condition.

The present invention is directed to methods and compositions that are effective in the inhibition of viral replication. In particular, the methods and compositions are effective at interfering with the activity of host cell proteins required in viral replication. For example, an embodiment of the invention is directed to methods and compositions comprising RNA sequences to which the host cell proteins TIAR and/or TIA-1 bind.

The present disclosure is directed to rapid detection and analysis of biological components using surface-enhanced Raman spectroscopy (SERS). One general aspect includes a method for preparing biological components from a biological sample for label-free SERS. The method also includes providing a Raman-background-free surfactant buffer to the biological sample. The method also includes agitating and heating the biological sample in the buffer to accelerate a lysis process thereby decomposing structural components of the biological sample. The structural components are substantially free of Raman background interference and are optimized for SERS signal enhancement.

Disclosed herein are immunoassay methods and reagents for detecting anti-Zika IgM antibody in a biological sample from a subject and/or diagnosing Zika virus infection in a subject. Also disclosed are algorithms for implementing the disclosed methods. The disclosed immunoassay methods, reagents, and algorithms enable efficient and reliable qualitative detection of anti-Zika virus antibodies and rapid determination of presumptive positive results for Zika virus infection in human subjects.