The present invention relates to a method for determining, predicting or estimating the biological age of a subject, or for providing a measurement for use in determining, predicting or estimating the biological age of a subject or for predicting the presence or absence of at least one disease in a subject, predicting the risk of a subject of having or developing at least one disease; and/or predicting the risk of mortality of a subject. This invention also relates to a device for determining the presence and/or amount of each biomarker in a set of biomarkers; a set of probes for determining the presence or amount of a set of biomarkers, and the use of such device and/or probes in any of the above methods. Also provided is a biomarker testing kit for use in a method as described herein and a computer-readable storage medium or a computer program comprising computer-executable instructions and associated method.

Methods and analyte sensors including at least a first working electrode having a first active area thereon, and performing a dip coating operation to deposit a bilayer membrane upon the first working electrode and the first active area. The bilayer may include an inner layer having a first membrane polymer and an outer layer having a second membrane polymer, the first membrane polymer and the second membrane polymer differing from one another. The dip coating operation may comprise one or more first dips in a first membrane formulation to form the inner layer of the bilayer membrane and one or more second dips in a second membrane formulation to form the outer layer of the bilayer membrane upon the inner layer.

Methods and analyte sensors including at least a first working electrode having a first active area thereon, and performing a dip coating operation to deposit a bilayer membrane upon the first working electrode and the first active area. The bilayer may include an inner layer having a first membrane polymer and an outer layer having a second membrane polymer, the first membrane polymer and the second membrane polymer differing from one another. The dip coating operation may comprise one or more first dips in a first membrane formulation to form the inner layer of the bilayer membrane and one or more second dips in a second membrane formulation to form the outer layer of the bilayer membrane upon the inner layer.

A system for identifying a plurality of biomarkers in a sample, including: a substrate including a plurality of microneedles projecting therefrom, each of the plurality of microneedles including a plurality of biomarker recognition molecules attached thereto, and the plurality of microneedles including a first microneedle and a second microneedle, the first microneedle including a first plurality of biomarker recognition molecules configured to recognize a first biomarker, and the second microneedle including a second plurality of biomarker recognition molecules configured to recognize a second biomarker different from the first biomarker.

Multiple enzymes may be present in the active area(s) of an electrochemical sensor to facilitate analysis of one or more analytes. The multiple enzymes may function independently to detect several analytes or in concert to detect a single analyte. One sensor configuration includes a first active area and a second active area, where the first active area has an oxidation-reduction potential that is sufficiently separated from the oxidation-reduction potential of the second active area to allow independent signal production. Some sensor configurations may have an active area overcoated with a multi-component membrane containing two or more different membrane polymers. Sensor configurations having multiple enzymes capable of interacting in concert include those in which a first enzyme converts an analyte into a first product and a second enzyme converts the first product into a second product, thereby generating a signal at a working electrode that is proportional to the analyte concentration.

Disclosed are assays, reagents, and methods for ultrasensitive detection of target molecules. The assay comprises fusion proteins including binding moieties, such as antibodies, nanobodies (VHH/VNAR), or aptamers, linked to nonfunctional fragments of a protein. In the presence of a target molecule, the binding moieties recognize distinct target regions and bring the protein fragments into close proximity, reconstituting a functional protein that generates a detectable signal. Linkers connecting the fusion components may be flexible peptides or polypeptides that spontaneously form dimers, trimers, or tetramers, thereby providing multivalent fusion proteins with enhanced sensitivity. The reconstituted protein may produce luminescent, fluorescent, colorimetric, or spectroscopic signals detectable by microplate readers, handheld luminometers, or lateral flow devices. The invention encompasses solid-phase, homogeneous, and lateral flow assay formats for detecting viruses, bacteria, proteins, peptides, or small molecules, including GLRaV-3, SARS-CoV-2, PSA, and E. coli. The disclosed assays exhibit improved specificity, reduced background, and enhanced signal-to-noise ratios.

A lateral flow assay for detecting a target in a liquid sample is disclosed. The lateral flow assay may comprise a first flow path and a second flow path for the liquid sample that merge at or before a membrane that generates a signal of immunoassay. Also disclosed are cartridges that include the lateral flow assay and methods for detecting a target in a liquid sample using the lateral flow assay.

Provided are a highly sensitive detection method for a target substance and a reagent for inspection thereof. Specifically, provided is a method of detecting a target substance that is a substance related to an enzyme reaction, the method including: mixing a specimen liquid that may contain the target substance and a reagent to obtain a liquid sample containing hydrogen peroxide, a peroxidase, a phenol derivative and a plurality of luminescent particles each having a binding functional group; aggregating the plurality of luminescent particles through binding of the binding functional groups of the luminescent particles based on a reaction of the hydrogen peroxide, the peroxidase and the phenol derivative in the liquid sample, which occurs when the target substance is present in the liquid sample; and obtaining a value related to fluorescence anisotropy of the liquid sample.