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Kit for rapidly detecting respiratory viruses and preparation method of kit

A technology of respiratory tract and detection test strips, which is applied in the field of medical diagnosis, can solve the problems of unsuitable on-site instant detection, unsuitable clinical screening, and long time consumption, so as to reduce the risk of crowd infection, save disposable consumables, and shorten the detection cycle. Effect

Pending Publication Date: 2017-06-13
SHANGHAI MUNICIPAL CENT FOR DISEASE CONTROL & PREVENTION +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0009] However, the above detection methods all require special equipment and professional training personnel, and can only be tested in a laboratory, which is expensive, complicated to operate, and time-consuming, and is not suitable for clinical screening, let alone on-site instant detection.
For example, virus culture is the gold standard for virus detection, but this detection method takes a long time, generally takes 3 to 7 days, and the sensitivity is not high
[0010] RT-PCR can detect samples within 4-8 hours, and has high sensitivity and specificity, but it is not suitable for airports, ports, railway stations, hospitals, etc. where the population density is high, the flow of people is large, and it is difficult for people to gather for a long time public places

Method used

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  • Kit for rapidly detecting respiratory viruses and preparation method of kit
  • Kit for rapidly detecting respiratory viruses and preparation method of kit
  • Kit for rapidly detecting respiratory viruses and preparation method of kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] The preparation of embodiment 1 respiratory virus rapid detection test paper

[0039] (1) Preparation of colloidal gold

[0040] The chemical reduction method is used to prepare the required colloidal gold particles by adding trisodium citrate reducing agent into the chloroauric acid aqueous solution. Method First, heat 500ml of 0.01% HAuCL4 solution to boiling, then quickly add 8.0ml of trisodium citrate aqueous solution, heat until red color appears, and then continue to boil for 10-15min. Restore volume to 500ml.

[0041] (2) Colloidal gold-antibody conjugate preparation

[0042] a. Take 500ml of colloidal gold solution, stir it with a magnetic stirrer, and use 0.2M K 2 The CO3 solution adjusted the pH to 8.0.

[0043] b. Dilute the mouse monoclonal antibody against respiratory syncytial virus (Tuno Company, Japan) to 1.5mg / ml with 0.01MPBS, take 5 centrifuge tubes and add 1ml colloidal gold solution respectively, add 30ul, 40ul, 45ul, 50ul of the antibody to be...

Embodiment 2

[0064] The detection application of embodiment 2 kit

[0065] This kit can qualitatively detect virus samples in nasal swabs, throat swabs, and nasal aspiration samples; through the detection of clinical samples, the coincidence rate of positive and negative is high, and it has high sensitivity and specificity.

[0066] 1180 clinical samples from patients with unknown source of infection were tested in three different hospitals, of which 410 were positive and 770 were negative.

[0067] Sensitivity: 399 positive samples were detected out of 410 positive samples, the sensitivity was 97.3%. Among them, 97 samples of respiratory syncytial virus, 114 samples of adenovirus, 85 samples of influenza A virus (FluA), and 103 samples of influenza B virus (FluB) were detected.

[0068] Specificity: 760 negative samples were detected out of 770 negative samples, the specificity was 98.7%.

Embodiment 3

[0069] Example 3: comparative study of adenovirus detection

[0070] Using the principle of immunochromatography, qualitative detection of adenovirus antigen in throat swab samples. Completely insert the sterilized cotton swab into the throat from the mouth, and rub it several times around the reddened part of the posterior pharyngeal wall and palatine tonsil to collect the mucosal epidermis. When collecting, be careful not to touch saliva.

[0071] Add 0.5ml of sample extraction solution (to the lower reference line) to the attached extraction tube in advance, and place it on the stand. Dip the sterilized cotton swab after collecting the sample into the sample extract in the extraction tube and stir. From the outside of the extraction tube, squeeze the sterilized cotton swab several times with your fingers to fully soak the sterilized cotton swab with the sample extract, then pull out the sterilized cotton swab, and the twisted liquid is used as the sample to be tested. Sque...

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Abstract

The invention provides colloidal gold method detection test paper for detecting respiratory viruses. The test paper provided by the invention comprises a sample pad, a glass fiber membrane containing a colloidal gold particle marker, a nitrocellulose membrane and water-absorbing paper, wherein the nitrocellulose membrane comprises a detection area and a control area; the detection area is formed by an influenza A virus antibody, an influenza B virus antibody, a respiratory syncytial virus antibody and an adenovirus antibody; and the control area is formed by an anti-mouse immune globulin antibody. If any one of or any combination of an respiratory syncytial virus, an adenovirus, an influenza A virus (FluA) and an influenza B virus (FluB) exists in a sample, an antigen is captured by the corresponding antibody immobilized by the detection area of the nitrocellulose membrane, and a claret-colored stripe is formed due to the action of colloidal gold. By adoption of the test paper provided by the invention, the respiratory syncytial virus, the adenovirus, the influenza A virus (FluA) and the influenza B virus (FluB) can be rapidly detected through one-time operation, so that the operating process is simplified and the aim of rapid detection is fulfilled.

Description

technical field [0001] The invention relates to the field of medical diagnosis, in particular to a multipurpose rapid detection kit for respiratory viruses and a preparation method thereof. [0002] Background of the invention [0003] It has been proven that 95% of acute upper respiratory diseases and most of the lower respiratory diseases are caused by pathogens other than bacteria, among which respiratory viruses are the most common, including respiratory syncytial virus (RSV) type A, type B, adenovirus ( ADV), influenza virus A (FluA), influenza virus B (FluB), parainfluenza virus 1 (PIV1), parainfluenza virus 2 (PIV2), parainfluenza virus 3 (PIV3), Coxsackie group B virus, etc. Among them, a variety of viruses have the characteristics of strong infectivity, fast transmission, short incubation period, and acute onset. Recently, new pathogenic viruses are still being discovered, such as human metapneumovirus, SARS virus, highly pathogenic poultry virus, etc. Influenza vir...

Claims

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Application Information

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IPC IPC(8): G01N33/558G01N33/543G01N33/569
CPCG01N33/558G01N33/543G01N33/56983
Inventor 陈健陈功祥吴寰宇朱国刚姜晨彦
Owner SHANGHAI MUNICIPAL CENT FOR DISEASE CONTROL & PREVENTION
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