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Quick test paper for detecting enterovirus and method for preparing same

A technology for detecting test strips and enteroviruses, which is applied in the field of rapid detection test strips for detecting enteroviruses and its manufacture, which can solve the problems of inability to distinguish infectious viruses from dead viruses, high costs, and backward detection methods, and achieve easy results. Read, strong specificity, and high sensitivity

Active Publication Date: 2014-04-02
万志静 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The virus isolation method is time-consuming and expensive; the immunofluorescence method requires expensive fluorescence microscopes and the specimens are not easy to preserve; the PCR method cannot distinguish between infectious viruses and dead viruses, and the above methods require professional operation
[0006] It is precisely because of the backwardness of existing detection methods that it is imperative to develop detection products that are easy to use, sensitive to detect, and low in price

Method used

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  • Quick test paper for detecting enterovirus and method for preparing same
  • Quick test paper for detecting enterovirus and method for preparing same
  • Quick test paper for detecting enterovirus and method for preparing same

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] Embodiment 1: Preparation of anti-EV71 virus monoclonal antibody

[0030] 1. Preparation of hybridoma cells

[0031] (1) SP2 / 0 myeloma cells were cultured in DMEM medium containing 10% calf serum for 48-72 hours, and when the cells grew well, they were ready for fusion.

[0032] (2) Antigen immunization: EV71 antigen 10ug was emulsified with an equal amount of complete Freund's adjuvant and injected subcutaneously for basic immunization. Three days before the fusion, 10 μg of the above-mentioned EV71 antigen was injected into the spleen and peritoneal cavity of the mouse, respectively.

[0033] (3) Preparation of immune spleen cells: the mice were sacrificed 3 days after the last booster immunization, and the spleen lymphocytes were collected aseptically.

[0034] (4) cell fusion

[0035] Fusion agent: PEG (4000); culture medium: DMEM with 10% calf serum. SP2 / O myeloma cells and lymphocytes of immunized BALB / C mice were fused at a ratio of 1:10 or 1:5.

[0036] (5)...

Embodiment 2

[0042] A kind of EV71 virus colloidal gold detection test paper, such as figure 1 As shown, it includes a bottom plate 1, a water-absorbing plate 7, a nitrocellulose membrane 2, a color particle storage pad 5, and a sample liquid-absorbing layer 6, wherein the color particle storage pad 5 is preferably a gold standard pad for EV71 virus monoclonal antibody; the middle part of the bottom plate is Nitrocellulose membrane. A detection line 3 and a polyclonal antibody control line 4 are arranged on the nitrocellulose membrane. One end of the bottom plate is a water-absorbing plate, and the other end is a sample liquid-absorbing layer. It overlaps and connects with the water-absorbing plate and the EV71 virus monoclonal antibody gold label pad, and a sample liquid-absorbing layer is pressed on the EV71 virus monoclonal antibody gold label pad, and the EV71 virus is detected by colloidal gold immunochromatography.

[0043] Test paper method:

[0044] 1. Two EV71 monoclonal antibodi...

Embodiment 3

[0055] A kind of EV71 virus latex detection test paper, comprises bottom plate 1, water-absorbing plate 7, nitrocellulose film 2, color granule storage pad 5, sample liquid-absorbing layer 6, wherein, color granule storage pad 5 is preferably EV71 monoclonal antibody latex particle storage Pad; the middle part of the bottom plate is a nitrocellulose membrane, on which a detection line 3 and a polyclonal antibody control line 4 are arranged, one end of the bottom plate is a water-absorbing plate, and the other end is a sample liquid-absorbing layer, and the nitrocellulose The two ends of the plain film are overlapped and connected with the water-absorbing board and the EV71 monoclonal antibody color particle storage pad respectively, and a sample liquid-absorbing layer is pressed on the EV71 monoclonal antibody color particle storage pad, and the EV71 virus is detected by latex chromatography.

[0056] Test paper method:

[0057] 1. Two EV71 monoclonal antibodies produced by th...

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Abstract

The invention discloses a quick test paper for detecting enterovirus and a method for preparing the same. The test paper detects enterovirus EV71 and Coxsackievirus A16 viruses in a sample by the immunochromatography adopting marking by colorful particles. The quick test strip is formed by sequentially overlapping and bonding a sample adsorption liquid layer, a colorful particle storing pad, a cellulose nitrate membrane and a water adsorption board on a bottom board, wherein the colorful particle storing pad is coated with a monoclonal antibody with a colorful particle mark; and the cellulose nitrate membrane is provided with a detection line sprayed by the monoclonal antibody of the EV71 and / or Coxsackievirus A16 and a control line sprayed by the polyclonal antibody of anti-mouse IgG. The test paper can quickly detects the EV71 and Coxsackievirus A16 viruses in the detected sample at the same time or respectively, finds the epidemic situation caused by the virus infection as early as possible and has the advantages of being easily operated, quickly getting results, avoiding special operators and the like.

Description

technical field [0001] The invention relates to a rapid detection test paper for detecting enterovirus and a manufacturing method thereof, in particular to a rapid detection test paper for detecting enterovirus EV71 and Coxsackievirus A16 viruses that can cause hand, foot and mouth disease. Background technique [0002] Hand foot mouth disease (Hand foot mouth disease, HFMD) is a global infectious disease, and there are reports of the prevalence of this disease in most parts of the world. Fever, rashes and ulcers on the hands, feet, mouth and other parts may cause myocarditis, pulmonary edema, aseptic meningoencephalitis and other complications in individual patients. There are more than 20 kinds of enteroviruses that cause HFMD, among which Coxsackievirus A16 (CoxA16) and enterovirus 71 (Enterovirus71.EV 71) are the most common. [0003] Australia, like the United States and Sweden, was one of the first countries to experience EV 71 infection. The EV 71 epidemic occurred ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/569G01N33/577G01N33/558
Inventor 万志静万志强
Owner 万志静
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