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Colloidal gold test strip for rapid detection of Human enterovirus 71 (EV71) IgM (immunoglobulin m)

A technology for colloidal gold test strips and enteroviruses, which is applied to measurement devices, instruments, scientific instruments, etc., can solve the problems of unsuitability for on-site measurement and long detection time, and achieves the effect of simple operation and guaranteed sensitivity.

Inactive Publication Date: 2012-09-19
蓝十字生物药业(北京)有限公司 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] Enzyme-linked immunoassay kits are mostly used to detect enterovirus type 71 (IgM) in the market. Although the detection limit of the kit is low (0.15mg / L), the detection time is long and it is not suitable for on-site determination. The application number is The invention patent application of 201010171862.2 proposed an enterovirus 71 antibody (IgM) diagnostic kit (enzyme-linked immunoassay), which needs to be tested before and after the detection of enterovirus 71 antibody (IgM). Instrument cleaning and maintenance work takes a long time to detect, and requires professional and technical personnel in the field to operate
[0008] At present, the combination of antibody capture method and immune colloidal gold technology can ensure the sensitivity and specificity of detection at the same time, and it is convenient to use, simple to operate, accurate and fast, and the colloidal gold test strip used to detect enterovirus 71 IgM is fresh There are reports

Method used

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  • Colloidal gold test strip for rapid detection of Human enterovirus 71 (EV71) IgM (immunoglobulin m)
  • Colloidal gold test strip for rapid detection of Human enterovirus 71 (EV71) IgM (immunoglobulin m)
  • Colloidal gold test strip for rapid detection of Human enterovirus 71 (EV71) IgM (immunoglobulin m)

Examples

Experimental program
Comparison scheme
Effect test

experiment example 1

[0046] Experimental example 1. the preparation method of test strip of the present invention

[0047] Reagents: EV71 monoclonal antibody, EV71 recombinant antigen Source: Ningbo Center for Disease Control and Prevention Anti-mouse IgG polyclonal antibody and anti-μ chain monoclonal antibody: Imported from Canada Artron BioResearch Inc.

[0048] The test strip preparation procedure is as follows

[0049] Step 1. Coating of cellulose film

[0050] 1. Use the anti-mouse IgG polyclonal antibody as the control line C, and the anti-μ chain monoclonal antibody as the detection line T. The two reagents are diluted with 20mM PBS according to the required concentration (range 1.5-2mg / ml).

[0051] 2. Coat the above prepared reagents on the nitrocellulose membrane according to the positions of C and T lines, and then dry for 6 hours at a temperature of 30°C and a humidity of ≤35% to prepare a cellulose membrane.

[0052] Step 2. Marking of the gold standard fiber layer:

[0053] 1. Ta...

Embodiment 2

[0066] Embodiment 2. Using method and methodological comparative experiment

[0067] How to use this product:

[0068] Serum / plasma / whole blood to be tested should be diluted 200 times, and dripped onto the sample fiber layer of the horizontal test strip, or immerse the part of the mucus end of the test strip below the MAX line in the diluted sample for 3 to 5 minutes, take it out and place it horizontally Wait for the color to develop.

[0069] Comparison 1: Product Comparison

[0070] Compared with the products of the prior art for detecting enterovirus 71 type IgM, the product of the present invention has the characteristics of fast, accurate, specific, and convenient, and does not need to do necessary instrument cleaning and cleaning before and after the detection of enterovirus 71 type IgM. Maintenance work, and does not require professionals in the field to operate.

[0071] Other products such as kits

[0072] Control product 1: an enterovirus 71 (EV71) IgM antibody...

Embodiment 3

[0099] Compared with an enterovirus 71 (EV71) IgM antibody detection test strip disclosed in the invention patent with application number 201110151009.9, the present invention has better specificity and higher positive rate.

[0100] The data list is shown in Table 3;

[0101] Negative and positive samples were tested 100 copies each

[0102] Pilot projects

Comparison product

Product of the present invention

EV71IgM positive samples (100 persons)

95%

100%

EV71 negative samples (100 persons)

94%

100%

[0103] In the present invention, the EV71 monoclonal antibody is selected for colloidal gold labeling, and then the EV71 IgM antibody in the sample is captured by the EV71 recombinant antigen, and the anti-μ chain antibody coated with the cellulose drug film can have a better specific binding reaction. The control product is carried out by colloidal gold labeling with anti-μ chain antibody, and EV71 antigen coated wi...

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Abstract

The invention discloses a colloidal gold test strip for rapid detection of Human enterovirus 71 (EV71) IgM (immunoglobulin m), belonging to the field of medical detection consumables. The colloidal gold test strip is characterized in that a colloidal gold-labeled cellulose pad is coated with colloidal gold-labeled EV71 antibody; a control line C is coated with anti-mouse IgG polyclonal antibody; a test line T is coated with anti-Mu chain monoclonal antibody; and a sample cellulose pad is coated with EV71 recombinant antigen. The inventive test strip adopts antibody capture principle and colloidal gold technology to detect EV71 IgM in serum / plasma / whole blood samples simply, sensitively, precisely and rapidly; and can be used for the rapid detection of EV71 IgM and diagnosis of hand-foot-mouth disease.

Description

[0001] technology field, [0002] The invention belongs to the field of biological detection, relates to a medical detection consumable, in particular to a colloidal gold test strip for rapid detection of enterovirus 71 IgM. Background technique [0003] Enterovirus type 71 IgM belongs to enterovirus type A. Enteroviruses are generally named by numbers, and the order of arrangement represents the order in which they were discovered. In sequence, this virus is named Human enterovirus 71 (Human enterovirus 71), or EV71 for short, and is one of the main pathogens that cause hand-foot and mouth disease (HFMD) in infants. Human enterovirus 71 was first isolated in 1969 from fecal samples of infants with central nervous system disease in California. [0004] EV71 mainly causes hand, foot and mouth disease, and can also cause various neurological diseases such as aseptic meningitis, brainstem encephalitis and polio-like paralysis. Hand, foot and mouth disease and central nervous sy...

Claims

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Application Information

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IPC IPC(8): G01N33/577
Inventor 张莹倪红霞孙茜
Owner 蓝十字生物药业(北京)有限公司
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