公开(公告)号：WO2022103282A1

公开(公告)日：2022-05-19

申请号：PCT/NZ2021/050201

申请日：2021-11-12

Applicant: INSITUGEN LIMITED

Inventor： HEATHER, Alison Kay

IPC: C12Q1/6897 ,  G01N33/74

Abstract: The present invention is concerned with the detection of ligands which bind to and activate steroid hormone receptors. Specifically, the present invention provides test kits and assay methods for the selective identification of steroid hormone receptor ligands from a test sample. Importantly, the test kits and assay methods described herein are cell-free, and do not require expensive-to-manufacture nuclear extracts for their performance. Instead, the test kits and assay methods described herein employ single polypeptide polymerases, such as T7 RNA polymerase, linked to a reporter construct. Activity of the enzyme is inhibited, rather than activated, by ligand-bound steroid hormone receptor complexes which only form in the presence of a target ligand. Accordingly, a measured change in a physical property of the reporter construct (e.g. fluorescence output) may be used to determine the presence of a target ligand in a sample under investigation.

公开(公告)号：WO2022103283A1

公开(公告)日：2022-05-19

申请号：PCT/NZ2021/050202

申请日：2021-11-12

Applicant: INSITUGEN LIMITED

Inventor： HEATHER, Alison Kay

IPC: C12Q1/6897 ,  G01N33/74 ,  C12N9/22

Abstract: The present invention provides diagnostic test kits and assay methods involving the detection of steroid hormone ligand(s) in a test sample. For example, in the detection of ligands which bind to and activate an androgen receptor, estrogen receptor, progesterone receptor, mineralocorticoid receptor and glucocorticoid receptor. In particular, the test kits and assay methods described herein are configured for field-based applications, for example, for trackside drug testing in athletes or at home testing of humans or animals for steroid hormone activity, which test kits, assays and methods colorimetric read-outs involving either inorganic pyrophosphate or CRISPR/Cas mediated RNA probe cleavage involving (e.g.) probes which have a detectable label including fluorophore/quencher molecules or fluorophore/tag molecules. Accordingly, the diagnostic test kits and assay methods described herein eliminate the requirement for detection instrumentation allowing reproducible 'rapid fire' tests in the field.