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Colloidal gold test paper for quickly detecting troponin I and preparation method for colloidal gold test paper

A technology of colloidal gold test paper and cardiac troponin, which is applied in the field of clinical immunology detection, can solve the problems of sensitivity restricting wide application, etc., and achieve the effect of improving detection sensitivity and expanding clinical application

Active Publication Date: 2012-12-26
北京乐普诊断科技股份有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The widely used gold standard immunoassay is an important means of detection of myocardial infarction, but its sensitivity limitation restricts further wider application

Method used

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  • Colloidal gold test paper for quickly detecting troponin I and preparation method for colloidal gold test paper
  • Colloidal gold test paper for quickly detecting troponin I and preparation method for colloidal gold test paper
  • Colloidal gold test paper for quickly detecting troponin I and preparation method for colloidal gold test paper

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0043] Embodiment 1 Cardiac troponin I colloidal gold test paper and preparation method thereof (referring to Fig. 1)

[0044] Preparation of cardiac troponin I antibody: commercial cTnI antibody (Hytest company, cat: 4T2) was selected, 10 mM PBS, pH 7.2, dialyzed overnight at 4°C.

[0045] Preparation of coated membrane (nitrocellulose membrane 2.5cm×0.6cm):

[0046] Preparation of coating buffer: 10 mM phosphate buffer solution with pH 7.2, filtered and sterilized by 0.22 μm microporous membrane, and stored at 4° C. for later use.

[0047] Dilute the anti-cardiac troponin I antibody and the rabbit anti-mouse antibody to 0.5mg / mL and 0.7mg / mL respectively with coating buffer, and use the Biodot three-dimensional spraying platform to spray the two at 1.5μL / cm Evenly spray-print on the nitrocellulose membrane with a width of 3.5cm, dry at 25-40°C for 8 hours, add a desiccant and seal it for later use.

[0048] Preparation of gold-labeled anti-cardiac troponin I monoclonal ant...

Embodiment 2

[0057] Embodiment 2 Cardiac troponin I colloidal gold test paper and preparation method thereof (referring to Fig. 1)

[0058] Preparation of coated membrane (nitrocellulose membrane 2.5cm×0.6cm):

[0059] Preparation of coating buffer: 10 mM phosphate buffer solution with pH 7.2, filtered and sterilized by 0.22 μm microporous membrane, and stored at 4° C. for later use.

[0060] The anti-cardiac troponin I antibody and the rabbit anti-mouse antibody (purchased from Abcam) prepared in Example 1 were diluted to 1.0 mg / mL and 1.5 mg / mL with coating buffer, respectively, and used the Biodot three-dimensional spraying platform to 1.5 μL The amount per cm was evenly spray-printed on a nitrocellulose membrane with a width of 3.5 cm at an interval of 1 cm, dried at 37 ° C for 8 hours, and sealed with a desiccant for later use.

[0061] Preparation of gold-labeled anti-cardiac troponin I monoclonal antibody gold-labeled pad 1

[0062] Use potassium carbonate to adjust the pH value o...

Embodiment 3

[0070] Embodiment 3 test strip sensitivity test of the present invention

[0071] Randomly extract 25 novel troponin I colloidal gold test papers made according to the above-mentioned embodiment 1 or 2, and 25 common commercially available troponin I colloidal gold test papers are used as a control group (pasting packs alternately on the bottom plate successively) film, colloidal gold pad sprayed with colloidal gold-labeled anti-cardiac troponin I monoclonal antibody, sample pad and absorbent pad). Dissolve cardiac troponin I ternary complex (SRM2921, purchased from Hytest Company) in troponin I-free serum (purchased from Hytest Company), and use the serum to dilute to 5 ng / mL, 1 ng / mL, 0.5 ng / mL, 0.1ng / mL, 5 strips were tested for each sample concentration. Table 1 shows the results of visual interpretation. It can be seen from the results that when the troponin I content in the serum is lower than 0.5ng / mL, the common colloidal gold test strip cannot be detected, and even...

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Abstract

The invention relates to the field of clinical immunology, particularly to colloidal gold test paper for quickly detecting troponin I and a preparation method for the colloidal gold test paper. The colloidal gold test paper comprises a coated film, a colloidal gold pad 1, a colloidal gold pad 2, a sample pad and a water absorbing pad which are mutually attached in a staggered way; the colloidal gold pad 1 is sprayed with a colloidal gold labeled cardiac troponin I resisting antibody; the colloidal gold pad 2 is sprayed with a colloidal gold labeled BSA (Bovine Serum Albumin)-resisting monoclonal antibody or PEG (Polyethylene Glycol)-resisting monoclonal antibody; and the coated film is coated with a cardiac troponin I resisting antibody detection line and a rabbit antimouse antibody quality control line. According to the improvement on the test paper strip, a PEG resisting technology and a BSA-resisting monoclonal antibody technology are introduced into the detection of the cardiac trooping I for the first time, so that the detection sensitivity of the cardiac trooping I is greatly improved and the clinical application of a traditional colloidal gold quick diagnosis test paper is enlarged.

Description

technical field [0001] The invention relates to the field of clinical immunology detection, in particular to a colloidal gold test strip for rapid detection of troponin I and a preparation method thereof. Background technique [0002] Myocardial infarction is one of the leading causes of death and disability worldwide. Globally, 17 million people die from cardiovascular diseases every year, and more than half of them die from acute myocardial infarction (AMI). In China, there are more than 20 million patients with coronary heart disease, and more than 1 million people are killed by acute myocardial infarction every year. Therefore, rapid identification and timely treatment of patients with early acute myocardial infarction are the key to improving their survival rate. For AMI patients without typical chest pain and ECG changes, the detection of serum myocardial markers is a necessary basis for the diagnosis of AMI. The criteria for myocardial infarction defined by the Wor...

Claims

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Application Information

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IPC IPC(8): G01N33/68G01N33/532
Inventor 余占江胡光宇王长青张芳
Owner 北京乐普诊断科技股份有限公司
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