Human respiratory tract pathogen flow cytometry detection kit and method and cell fixation solution

A detection kit and cell fixation technology, applied in the field of pathogen diagnosis, can solve the problems of technologies and products that have not yet been seen to detect respiratory pathogens, and achieve the effects of saving resources and time, objective and accurate results, and saving labor costs

Active Publication Date: 2016-03-09
GUANGDONG HECIN SCI INC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Since flow cytometry uses cell surface factors as the detection object, and human respiratory pathogens are generally located inside the cells, so far there is no technology and product that can make good use of flow cytometry to detect respiratory pathogens

Method used

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  • Human respiratory tract pathogen flow cytometry detection kit and method and cell fixation solution
  • Human respiratory tract pathogen flow cytometry detection kit and method and cell fixation solution
  • Human respiratory tract pathogen flow cytometry detection kit and method and cell fixation solution

Examples

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Embodiment 1

[0031] Example 1 cell fixative

[0032] The cell fixative solution described in this embodiment contains a PBS solution with a volume fraction of 0.5% formaldehyde and a volume fraction of 60% methanol. The preparation method is as follows: adding formaldehyde and methanol to PBS solution, so that the volume fraction of formaldehyde is 0.5%, and the volume fraction of methanol is 60%, and the pH value is adjusted to 7.3 to obtain the product.

Embodiment 2

[0033] Embodiment 2: the flow cytometric detection kit of human respiratory tract pathogen

[0034] This embodiment provides a flow cytometric detection kit for human respiratory pathogen infection. The kit includes the cell fixative described in Example 1, cell permeabilization agent and fluorescein (FITC-fluorescein isothiocyanate) labeled monoclonal antibody against Adv antigen (purchased from Guangzhou Ruida Biotechnology Co., Ltd. ), flow cytometry cell washing solution. In this embodiment, the cell permeation agent is Triton-X100 solution with a volume fraction of 1%. The cell washing solution is PBST (1×, pH7.4)

Embodiment 3

[0035] Embodiment 3: the flow cytometry detection method of human respiratory tract pathogen

[0036] This embodiment is a method for detecting a single pathogen (Adv) infection using the flow cytometry kit in Example 2, and proceeds as follows:

[0037] 1. Bronchoalveolar lavage fluid sample collection and cell fixation: collected by professional medical staff, after sampling, take 3ml into the sampling tube. The sampling tube was centrifuged at 500 g for 8 minutes, the supernatant was discarded, and the cell layer was not sucked away. Add 3ml of cell fixative and resuspend the cells, and tighten the tube cap. Store the sampling tube upright at -20°C.

[0038] 2. Preparation of single cell suspension: place the above-mentioned sampling tube containing the specimen on a shaker for 10-15 seconds, take out and discard the swab. Centrifuge the above-mentioned sampling tube at 500g for 8 minutes, discard the supernatant, and be careful not to suck away the cell layer. Then add...

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Abstract

The invention provides a cell fixation solution, detection kit and method for human respiratory tract pathogen infection flow cytometry detection. The cell fixation solution is a PBS solution containing formaldehyde with the volume fraction of 0.1-1% and methyl alcohol with the volume fraction of 60-80%, and the pH of the cell fixation solution is 7.2-7.4. The kit comprises the cell fixation solution, a cell permeating agent and at least one kind of monoclonal antibodies of a human respiratory tract pathogen to be detected, each kind of monoclonal antibodies of the human respiratory tract pathogen to be detected is marked with fluorescein, and different human respiratory tract pathogens to be detected are provided with different fluoresceins. The detection process is automatic, labor cost is saved, the result is objective and accurate, single-sample multiple detection can be achieved, and the pathogen infection condition can be better reflected.

Description

technical field [0001] The invention belongs to the technical field of pathogen diagnosis, and in particular relates to a flow cytometry detection method, a kit and a cell fixative for human respiratory pathogens. Background technique [0002] Influenza A (IA), Influenza B (IB), Respiratory Syncytial Virus (RSV) and Respiratory Adenovirus (ADV), Parainfluenza 1, 2, 3, Metapneumovirus, Bocavirus, Coronavirus, Rhinovirus , enteroviruses, mycoplasma, and chlamydia are the main pathogens that cause respiratory diseases. Among them, influenza virus A, influenza virus B, respiratory syncytial virus and respiratory adenovirus are the most common, accounting for about 50% of the viral pathogens of respiratory diseases. For more rapid and sensitive detection of respiratory viruses, rational use of various antibiotics, and use of new antiviral drugs for the treatment of influenza, early detection and identification of infectious viruses has become very important. [0003] The curren...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/569G01N33/533
CPCG01N33/533G01N33/56927G01N33/56933G01N33/56983
Inventor 李小锋李晨阳刘铭龙
Owner GUANGDONG HECIN SCI INC
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