摘要:
An enzymatic method for the analytical determination of creatine kinase in an aqueous liquid such as blood serum is described. The determination is made by measuring an optical density change using the reagents creatine phosphate, adenosine diphosphate, glycerol, glycerol kinase, .alpha.-glycerophosphate oxidase, a chromagen and a mercapto-containing creatine kinase activator. The mercapto-containing creatine kinase activator is added in encapsulated form or in low concentrations so as to preserve the activity of the chromagen.
摘要:
An aqueous composition containing primers for opposing strands of a target retroviral DNA (such as HIV-I DNA) can be used in polymerase chain reaction to provide simultaneously rapid and efficient amplification and detection of that target DNA and one or more additional target DNA's. The primers for each target DNA differ in length by no more than 5 nucleotides and have a T.sub.m within the range of from about 65.degree. to about 74.degree. C., while all of the T.sub.m 's are within about 5.degree. C. of each other. Such compositions are useful in diagnostic test kits and methods for amplification and detection of multiple nucleic acids, or in "multiplexing", using multiple capture probes, all of which have T.sub.m 's which are greater than 50.degree. C. and within 15.degree. C. of each other.
摘要:
Specific binding methods are used for diagnostic assays and purification separations whereby the specific binding capture reagent is prepared from copolymers having highly reactive carboxy groups. These groups are extended from the polymer surface with a linking group having from 8 to 50 atoms in the chain and two or more alkylene, arylene, alkylenearylene or arylenealkylene groups. To these reactive groups is attached a biologically active substance such as a protein or oligonucleotide which then participates in the diagnostic assays or purification separation methods.
摘要:
A cuvette and a method of use which prevent nucleic acid amplified by PCR technology from being released to the atmosphere, while still proceeding to a detection step to determine whether or not the nucleic acid is present. Detection reagents are either pre-incorporated into compartments in the cuvette or added after amplification. In the latter case, a check valve prevents amplified nucleic acid from being released. Transfer of liquids between compartments is achieved via the use of flexible compartment walls and an external pressure source, or via pistons that are part of the cuvette and operate on the compartments as a piston within a piston chamber.
摘要:
Nucleic acids can be made available for amplification or other treatment after lysis by contacting the lysate with specific weakly basic polymers to form a precipitate with the nucleic acids at acidic pH. After removing non-precipitated materials, the pH is then made basic, thereby releasing the nucleic acids from the polymer. This method for preparing specimen samples is simple and quite rapid, and the released nucleic acids can be further treated in hybridization assays or amplification procedures. The weakly basic polymers are water-soluble and cationic at acidic pH, but neutral in charge at basic pH.
摘要:
An oligonucleotide is linked to a particle through a protein or carbohydrate to form a water-insoluble nucleic acid probe. The protein or carbohydrate has a pI of about 6 or less, and has been chemically modified with an acylating, alkylating or sulfonylating agent. The particle surface is substantially free of other proteins or carbohydrates. The probe is useful in various diagnostic and purification methods wherein hybridization of the oligonucleotide with a target nucleic acid is possible. In one instance, the probe can be used to capture a DNA strand which has been amplified using polymerase chain reaction techniques.
摘要:
Biologically active reagents are prepared from particles of copolymers having highly reactive carboxy or equivalent groups. The reagents are prepared by covalently attaching biologically active substances, for example antibodies, to the particles, directly or indirectly through highly reactive carboxy groups on the particle surface. These reagents are used to advantage in analytical elements, methods for the detection of specific binding ligands (such as immunological species) and immunoassays, and in purification methods as affinity chromatography reagents.
摘要:
A multilayer analytical element for the determination of a clinically significant enzyme analyte comprises a photosensitive compound (e.g. a photosensitive dye or dye precursor) and a filter layer containing one or more filter dyes. The filter layer is situated in the element such that incident radiation used to detect a density change resulting from interaction of the analyte and the photosensitive compound passes through the filter layer before it reaches the photosensitive compound. The use of the filter layer inhibits premature changes in the photosensitive compound caused by incident radiation. This element is particularly useful for the determination of creatine kinase or one of its isoenzymes, e.g. creatine kinase-MB.
摘要:
A nucleic acid test article can be used to detect a targeted nucleic acid found in a specimen. The test article includes a substrate having two opposing surfaces and a water-insoluble nucleic acid probe attached in a distinct zone of one of the surfaces. The probe is prepared from a water-insoluble particle to which is covalently attached an oligonucleotide which is complementary to the targeted nucleic acid. Substantially none of the probe is embedded within the surface of the substrate. Particularly useful test articles have a multiplicity of water-insoluble probes located in distinct zones on one of the substrate surfaces. These probes are useful for the detection of a multiplicity of targeted nucleic acids, particularly after amplification by polymerase chain reaction.
摘要:
A device and method are disclosed for amplifying and detecting nucleic acid material. The device and method use a label and signalling material responsive to the label to produce a detectable signal. A surprising result of the method and device is that at least one of the wash steps heretofore required has been eliminated without substantially adversely affecting the results.