A nucleic acid probe capable of participating in a chemiluminescent reaction comprising a defined nucleic acid sequence, the sequence being linked to any one of a. a chemiluminescence precursor, b. a chemiluminescence enhancer, and c. an enzyme the remaining two of (a), (b) and (c) not linked to the sequence being in a mixture of the linked sequence. A method for determining a particular single stranded polynucleotide sequence in a test medium, comprising the steps of: (a) combining the test medium with a polynucleotide probe having a base sequence substantially complementary to the sequence to be determined, (b) labeling either the resulting hybrids or probe which has not hybridized with the sequence to be determined with one of the participants in an enhanced chamiluminescent reaction involving a chemiluminescent precursor, an enzyme, an oxidant, and a chemiluminescence enhancer, (c) initiating such chemiluminent reaction with the labeled hybrid or probe, and (d) detecting the resulting light emission.
This invention discloses a method for coevolving products from two or more reactants, along with the nucleic acid that can facilitate the reaction for making the products. The invention further discloses the products and facilitating nucleic acids produced by said method.
A method and test kit using a hydroxyaryl cyclic diacylhydrazide, a peroxide and a peroxidase enzyme on a blotting membrane for detecting DNA, RNA or proteins (polypeptides) is described. The method and kit provides enhanced chemiluminescence in an assay because of the use of the membrane. The method and test kit is particularly useful in Western, Southern and Northern blotting type assays and DNA sequencing.
Probes labeled with 1,2-dioxetane precursors can be employed in a variety of assays. The probes may be nucleic acid, peptide nucleic acid, proteins including enzyme, antibody or antigen, steroid, carbohydrate, drug or non-drug hapten. The probe is provided with a 1,2-dioxetane precursor bound thereto, generally either covalently, or a strong ligand bond. The dioxetane precursor moiety is converted to a bound 1,2-dioxetane by exposure to singlet oxygen. These dioxetane (labels) either spontaneously decompose, or are induced to decompose by an appropriate trigger to release light. The trigger may be a change in pH temperature, or an agent which removes a protective group. Assay formats in which these 1,2-dioxetane labeled probes and referents may be used to include hybridization assays, immuno assays, gel-based assays and Capillary Zone Electrophoresis.
An assay method, compositions and test kits using a hydroxyaryl cyclic diacylhydrazide is described. A hydrogen peroxide and peroxidase enzyme. The preferred compositions incorporate enhancer compounds and a chelating agent which suppresses light production prior to addition of a peroxidase enzyme. The assay method can test for a peroxidase enzyme, a peroxide or can be used in immunoassays and probe assays.
A hybridization carrier, containing a single-stranded polynucleotide having the formula: wherein N represents admine, guanine or cytosin; T represents thymine; n is an integer of 2 or larger; and m is an integer of 5 or larger; the polynucleotide being immobilized by an amide bond on a surface of an organic polymers particle having a diameter of from about 0.05 .mu.m to about 5 .mu.m; the polynucleotide being immobilized at the site of a nucleotide sequence consisting of 2 or more polynucleotide which contain a primary amino residue in the polynucleotide; and the amide bond having been formed between the primary amino residue and a carboxyl residue on the surface of the organic polymer particle.
