Human heparin binding protein immunoturbidimetry detection reagent and application thereof

A heparin-binding protein and reagent technology, which is applied in the field of immunoassay, and can solve the problems of large venous blood volume, inability to detect, and inability to analyze and measure by analytical instruments.

Pending Publication Date: 2021-07-30
苏州优诺康生物技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In order to obtain serum or plasma samples, a large amount of venous blood needs to be collected, and after blood collection, inspectors need to pretreat the whole blood sample, which cannot be directly tested, which makes it impossible for analytical instruments to perform fully automatic analysis relying on existing kits Determination, and the entire detection process takes a long time

Method used

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  • Human heparin binding protein immunoturbidimetry detection reagent and application thereof
  • Human heparin binding protein immunoturbidimetry detection reagent and application thereof
  • Human heparin binding protein immunoturbidimetry detection reagent and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0080] Embodiment 1, preparation reagent

[0081] 1.1 Preparation of pretreatment reagents

[0082] Take deionized distilled water to dissolve sodium chloride, add PB buffer and mix evenly, then add a small amount of TritonX-100, PEG6000 and proclin300, adjust the concentration and pH value, and prepare the pretreatment reagent. In the pretreatment reagent, the concentration of PB buffer was 25mM, the concentration of TritonX-100 was 2.0mg / mL, the concentration of sodium chloride was 40mg / mL, the concentration of PEG6000 was 20mg / mL, and the concentration of proclin300 was 0.3mg / mL. The pH of the pretreatment reagent was 6.0.

[0083] 1.2 Preparation of recognition reagents

[0084] Recognition reagents include latex microspheres coupled with HBP antibody, PB buffer, sodium chloride, bovine serum albumin, Tween 20, and proclin300. In the recognition reagent, the concentration of PB buffer is 50mM, the concentration of latex microspheres coupled with HBP antibody is 10mg / mL,...

Embodiment 2

[0093] Embodiment 2, make standard curve

[0094] 2.1 Determination of the absorbance of the calibrator

[0095] The absorbance (OD) of the HBP antigen calibrator was measured on an automatic protein analyzer using the pretreatment reagent, recognition reagent and 9 different concentrations of the HBP antigen calibrator prepared in Example 1, and the measurement conditions are shown in Table 1. The HBP antigen calibrator of each concentration was measured three times and the average value was taken, and the measurement results are shown in Table 2.

[0096] Table 1. Assay conditions

[0097] temperature 37℃ Test wavelength 650nm Sample load 5ul Pretreatment Reagent Loading Amount 150ul Identify reagent load 50ul Reaction time 3min

[0098] Table 2. Absorbance measurement results of different concentrations of HBP antigen calibrator

[0099] Concentration (ng / mL) 0 25 50 100 200 400 600 800 1000 Absorbanc...

Embodiment 3

[0102] Embodiment 3, measure minimum detection limit (analysis sensitivity)

[0103] 3.1 Repeatedly measure the absorbance of the blank sample

[0104] Using the zero-value HBP antigen calibrator selected in Example 1 as a blank sample, using the pretreatment reagent and identification reagent prepared in Example 1, the absorbance of the blank sample was measured on an automatic protein analyzer. Refer to Example 2 for the measurement conditions, repeat the measurement 10 times, and see Table 3 for the measurement results.

[0105] Table 3. Absorbance measurement results of blank samples

[0106] Assay number 1 2 3 4 5 6 7 8 9 10 Absorbance (OD) 253 248 229 261 258 244 258 249 241 254

[0107] 3.2 Calculate the minimum detection limit

[0108] Calculate the mean absorbance of the blank sample and the standard deviation (SD) of the absorbance of the blank sample, was 249.5 and SD was 9.63. by Substituting the values ​​into the HB...

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Abstract

The embodiment of the invention discloses a kit for detecting heparin binding protein in a sample. The kit comprises a pretreatment reagent for pretreating a sample and a recognition reagent for recognizing heparin binding protein. The pretreatment reagent comprises a hemolytic agent and an electrolyte, the hemolytic agent is TritonX-100 with the concentration of 1-3 mg / mL, the electrolyte is sodium chloride with the concentration of 30-60 mg / mL, and the pH value of the pretreatment reagent is 5.5-6.5. The kit can be used for detecting the concentration of the heparin binding protein by using a whole blood sample, the blood sampling amount required for detection is small, and the detection time is short. The embodiment of the invention further discloses application of the kit in quantitative detection of the heparin binding protein and a method.

Description

technical field [0001] This specification relates to the field of immunoassay, in particular to a human heparin-binding protein immunoturbidimetric detection reagent and its application. Background technique [0002] Heparin-binding protein (HBP), also known as CAP37 or azurecin, is a granular protein with various functions released by neutrophils upon external stimulation. HBP can activate monocytes and macrophages, has significant antibacterial activity, chemotactic properties, and promotes vascular leakage, making it an emerging infectious disease-related biomarker. Once a bacterial infection occurs, the infection-related M protein combines with fibrinogen, resulting in the activation of multilineage leukocytes and the secretion of a large amount of HBP. HBP is widely used in the diagnosis and differential diagnosis of clinical infectious diseases, especially in the early diagnosis and curative effect monitoring of patients with sepsis, and also in other infectious disea...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68G01N33/539
CPCG01N33/539G01N33/6893G01N2800/26
Inventor 顾金华梁佳然
Owner 苏州优诺康生物技术有限公司
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