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Human serum amyloid protein A determination kit with high sensitivity and wide detection range

A technology of amyloid protein and human serum, which is applied in the field of immunoassay, can solve the problems of insufficient reagent detection sensitivity, insufficient sensitivity, and insufficient linear range, etc.

Inactive Publication Date: 2019-04-16
BEIJING BEIER BIOENG
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Chromatography reagent system varies greatly, quantification is not accurate enough, sensitivity is not enough; latex turbidimetry reagent can realize automatic mode operation on the biochemical analyzer, the system error is small, the precision is good, the reagent stability is good, but the existing reagent The detection sensitivity is not good enough and the linear range is not wide enough

Method used

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  • Human serum amyloid protein A determination kit with high sensitivity and wide detection range
  • Human serum amyloid protein A determination kit with high sensitivity and wide detection range
  • Human serum amyloid protein A determination kit with high sensitivity and wide detection range

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0061] A human serum amyloid A assay kit, comprising reagent 1, reagent 2 and the SAA calibrator of different concentration gradients; wherein,

[0062] Reagent 1 includes the following final concentrations of components: 20 mM HEPES buffer (pH 7.2), 0.5% (w / v) of BSA, 3% (w / v) of PEG6000, 1% (w / v) of Mw Be the dextran of 40000, the KCl of 150mM, the Tween-20 of 0.05% (w / v), the ProClin300 of 0.1% (w / v), the EDTA of 2mM;

[0063] Reagent 2 includes the components of the following final concentrations: carboxyl latex microspheres (average particle diameter is 240nm, microsphere final concentration is 0.02% (w / v)), 0.0065% (w / v ) labeled human SAA monoclonal antibody, carboxyl latex microspheres labeled only with human SAA polyclonal antibody (average particle size is 123nm, final concentration of microspheres is 0.2% (w / v)), 0.04% (w / v) Labeled human SAA polyclonal antibody, 50mM MOPS buffer (pH7.4), 100mM potassium chloride, 0.3% (w / v) gelatin, 1% (w / v) BSA, 2% (w / v) sucrose ...

Embodiment 2

[0070] A human serum amyloid A assay kit, comprising reagent 1, reagent 2 and the SAA calibrator of different concentration gradients; wherein,

[0071] Reagent 1 included the following final concentrations of components: 50 mM MOPS buffer (pH 7.4), 1.5% (w / v) of BSA, 2% (w / v) of PEG8000, 5% (w / v) of Mw 5000 polylysine, 300mM NaCl, 0.05% (w / v) Tween-80, 0.1% (w / v) ProClin300, 2mM EDTA;

[0072] Reagent 2 includes components of the following final concentrations: carboxyl latex microspheres (average particle diameter of 273 nm, final microsphere concentration of 0.01% (w / v)) labeled with human SAA monoclonal antibody, 0.005% (w / v ) labeled human SAA monoclonal antibody, carboxyl latex microspheres labeled only with human SAA polyclonal antibody (average particle size is 70nm, final concentration of microspheres is 0.3% (w / v)), 0.065% (w / v) Labeled human SAA polyclonal antibody, 50mM HEPES buffer (pH7.2), 100mM ammonium chloride, 0.2% (w / v) gelatin, 0.5% (w / v) BSA, 3% (w / v) suc...

Embodiment 3

[0079] A human serum amyloid A assay kit, comprising reagent 1, reagent 2 and SAA calibrator with different concentration gradients; wherein,

[0080] Reagent 1 includes the following final concentrations of components: 50 mM PBS buffer (pH 7.4), 2% (w / v) of BSA, 1% (w / v) of PEG20000, 2% (w / v) of Mw 10000 PVP, 100mM NH 4Cl, 0.1% (w / v) Tween-80, 0.1% (w / v) ProClin300, 10 mM EDTA;

[0081] Reagent 2 includes the components of the following final concentrations: carboxyl latex microspheres (average particle diameter is 223nm, microsphere final concentration is 0.015% (w / v)), 0.0072% (w / v ) labeled human SAA monoclonal antibody, carboxyl latex microspheres labeled only human SAA polyclonal antibody (the average particle size is 83nm, the final concentration of microspheres is 0.25% (w / v)), 0.055% (w / v) Labeled human SAA polyclonal antibody, 50mM TAPS buffer (pH8.0), 50mM magnesium chloride, 0.2% (w / v) gelatin, 0.2% (w / v) BSA, 3% (w / v) sucrose, 1 % (w / v) glucose, 0.05% (w / v) Twe...

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Abstract

The embodiment of the invention relates to the field of immunodetection, in particular to a human serum amyloid protein A determination kit with high sensitivity and a wide detection range. The humanserum amyloid protein A determination kit comprises a reagent 1, a reagent 2 and SAA calibrators with different concentration gradients, wherein the reagent 2 comprises carboxylated latex microspheresonly labeled with human SAA monoclonal antibodies and carboxylated latex microspheres only labeled with human SAA polyclonal antibodies; the average particle size of the carboxylated latex microspheres only labeled with the human SAA monoclonal antibodies is larger than the average particle size of the carboxylated latex microspheres only labeled with the human SAA polyclonal antibodies. The kitcan be used for detecting a single sample within 10 minutes; the kit has the advantages that the kit is simple in structure and excellent in precision, accuracy, linearity, interference resistance andother indexes, can well diagnose early inflammation of infectious diseases, can detect the content of SAA in serum and the content of SAA in blood plasma, is very suitable for clinical biochemical analyzers, and greatly facilitates clinical examination.

Description

technical field [0001] The invention relates to the field of immunoassay, in particular to a human serum amyloid A assay kit with high sensitivity and wide detection range. Background technique [0002] Serum amyloid A (serum smyloid A, SAA) is a heterogeneous protein in the apolipoprotein family. It is an acute phase response protein with a relative molecular weight of about 12KD. Its function is similar to that of the third component of high-density lipoprotein (HDL3 ), the affinity of macrophages for HDL-SAA conjugates is 2-3 times higher than that for HDL alone. There are four subtypes of SAA, among which SAA1 and SAA2 are acute proteins, which rise rapidly during the acute phase of inflammation; SAA3 gene cannot be expressed in humans; SAA4 can be continuously expressed, but does not participate in acute inflammatory response. According to the expression in vivo, SAA can be divided into acute SAA (acuteSAA, A-SAA) and constitutive SAA (constitutive SAA, C-SAA). SAA in...

Claims

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Application Information

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IPC IPC(8): G01N33/68G01N33/543G01N33/532
CPCG01N33/532G01N33/5432G01N33/68G01N2800/7095
Inventor 不公告发明人
Owner BEIJING BEIER BIOENG
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