Lateral flow based methods and assays for rapid and inexpensive diagnostic tests

摘要

The invention provides reagents and methods for lateral flow assays and quantitative capture or determination of components, including cells, in a sample. In one aspect, reagents and methods for diagnostic assay are provided. In one embodiment an assay for determining T cell numbers, particularly a CD2+ CD4+ T cell assay is provided. A manufacturing method for producing rapid diagnostic assays in a decentralized manner is also described. The method generates net economic advantages over conventional diagnostic manufacturing practices.

主权项

1. A system for the detection and quantitation of cells of interest in a sample, said system comprising:
(A) a solid support which is a wickable medium suitable for the reception, lateral flow and transport of said sample and any cells therein, wherein the medium supports cells moving over the pre-wetted surface propelled by liquid flow pressure; (B) a scaffold or polymer having a repeating unit, which scaffold or polymer is bound covalently or non covalently to the solid support of (A) and applied to the solid support in bands of varying concentrations or dilutions after a first capture area, wherein the scaffold or polymer is a nucleic acid having defined or repeating nucleotide sequence and to which multiple antibodies, peptides or other binding agents are affixed; (C) a first capture reagent capable of binding directly or indirectly with the cells of interest in the sample, which first reagent is bound covalently or non covalently to the support of (A) in a capture area, wherein the first capture reagent is one or more antibody, antigen, peptide, protein or ligand configured to bind to an antigen expressed by the cells of interest; (D) a surrogate polymer, which is a nucleic acid having defined or repeating complementary nucleotide sequence configured to hybridize with the nucleic acid scaffold or polymer of (B), and which acts as a surrogate marker and is configured for lateral flow in the solid support and for binding directly or indirectly with the cells of interest in the sample; and (E) a non-radioactive indicator configured to indicate an amount of scaffold or polymer of (B) which is bound in an assay; whereby a presence and the amount of cells of interest in the sample is indicated by the depletion of the surrogate marker such that less surrogate marker is available for binding with the scaffold or polymer of (B) and the intensity of the indicator is reduced.