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Latex enhanced immunoturbidimetric detection kit of cardiac troponin T

A technology for cardiac troponin and detection kits, which is applied in the fields of medicine, immunity and in vitro diagnosis, and can solve the problems of batch-to-batch difference, high repeatability and high reagent cost

Active Publication Date: 2019-05-17
BEIJING STRONG BIOTECH INC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The main disadvantages of using ELISA to detect cTnT are the need for washing and separation steps during the determination process, time-consuming, low degree of automation, batch-to-batch variation and relatively large repeatability
Although the detection of cTnT by chemiluminescence has high sensitivity and linearity, the cost of reagents is relatively high and the supporting large-scale instruments are relatively expensive

Method used

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  • Latex enhanced immunoturbidimetric detection kit of cardiac troponin T
  • Latex enhanced immunoturbidimetric detection kit of cardiac troponin T
  • Latex enhanced immunoturbidimetric detection kit of cardiac troponin T

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0050] Example 1. Preparation method of latex particles bound to antibodies

[0051] (1) Rinse 10mg / mL latex particles twice with 10mL MES buffer (average particle size 300nm, purchased from JSR) (average particle size 200nm and average particle size 400nm particles can be used instead);

[0052] (2) Resuspend the particles in 10 mL of active buffer (10 mM MES, pH 6.0) to make the particle concentration 0.1% w / v, and use ultrasound or stirring to ensure that the particles are well resuspended;

[0053](3) Add 5 mg / mL EDC (the amount of EDC is 0.5 to 2.5 times the carboxyl content on the latex) to the granules in step 2, react at 25°C to 45°C for 15 to 30 minutes, keep mixing, and activate;

[0054] (4) Rinse the activated particles twice with 10mM boric acid buffer pH9.0, and resuspend in 5mL boric acid buffer (10mM, pH9.0);

[0055] (5) Dissolving the cTnT antibody (mouse monoclonal antibody) in 5 mL of borate buffer;

[0056] (6) Mix the particle suspension in step 4 and t...

Embodiment 2

[0059] Embodiment 2. The preparation method of the kit of the present disclosure

[0060] Prepare reagents according to the following component contents:

[0061] 1. The composition of the first reagent:

[0062] Gly buffer 100mM

[0063] Bovine Serum Albumin 0.5% w / v

[0064] Biolipidure 405 0.1% (or 0.2% w / v of biolipidure 103)

[0065] NaN 3 0.1%w / v

[0066] Its pH is 7.5.

[0067] 2. The composition of the second reagent:

[0068] Gly buffer 100mM;

[0069] cTnT antibody-coated latex particles 0.1% w / v;

[0070] Bovine serum albumin 0.5% w / v;

[0071] NaN 3 0.1%w / v

[0072] Its pH is 8.0.

[0073] 3. Preparation of calibrator:

[0074] Calibrator can be prepared by yourself: Dissolve commercially available human cTnT recombinant protein (purchased commercial antigen) in diluent (pH7.4, NaCl 0.9%, bovine serum albumin 0.5%, NaN 3 0.1%) to make 6 concentrations of calibrator (0, 1.5, 3, 6, 12, 24ng / mL). Commercially available cTnT calibrators can also be us...

Embodiment 3

[0075] Embodiment 3. Preparation of contrast kit

[0076] According to the method of Example 2, the only difference is that Biolipidure is replaced by PEG-12000 at a concentration of 0.5% w / v.

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Abstract

Theinvention relates to a latex enhanced immunoturbidimetric detection kit of cardiac troponin T.More precisely, the latex enhanced immunoturbidimetric detection kit comprises a first reagent and a second reagent, wherein the first reagent comprises a buffer solution, a stabilizer, a copolymer of 2-methyl allyl oxyethyl phosphonyl choline, and a preservative; and the second reagent comprises the buffer solution, latex granules bound with anti-human troponin T antibody, the stabilizer, and the preservative. The latex enhanced immunoturbidimetric detection kit of the cardiac troponin Thas a quantitative detection limit of about 0.1 ng / ml, the correlation with commercial chemiluminescence reagent is good, and can be applied to the clinical test of cTnT.

Description

technical field [0001] The disclosure relates to the fields of medicine, immunity and in vitro diagnosis, and specifically relates to a kit for detecting troponin T by immunoturbidimetry. Background technique [0002] Troponin T (TnT) has a molecular weight of 37 kD and is a tropomyosin-binding subunit. There are three subtypes of TnT, among which skeletal muscle troponin T (sTnT) includes fast skeletal muscle type and slow skeletal muscle type; in addition, there is cardiac muscle type (cTnT). cTnT has 40% non-homology relative to the two skeletal muscle subtypes. [0003] In view of cTnT molecular stability, hydrophilicity, specific antigenic determinant and good reactivity, it can be used as a specific marker of myocardial injury. The serum half-life of cTnT is only 120 minutes, but the elevated serum concentration of cTnT can last for several days to three weeks, up to 27 days in patients with acute myocardial infarction (AMI). When cardiomyocytes are reversibly damag...

Claims

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Application Information

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IPC IPC(8): G01N33/68G01N33/531G01N35/00G01N21/31
Inventor 韩成莲刘瑶刘希
Owner BEIJING STRONG BIOTECH INC
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