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Reagent for Respiratory Virus Treatment and Its Application

A respiratory and virus technology, applied in the field of respiratory virus treatment reagents, can solve the problems affecting the stability of viral nucleic acid, the impact of results, and the impact of screening efficiency, so as to save transshipment, reduce the level of protection, and improve screening efficiency.

Active Publication Date: 2021-06-04
THE FIRST AFFILIATED HOSPITAL OF GUANGZHOU MEDICAL UNIV (GUANGZHOU RESPIRATORY CENT) +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

For example, this SARS-CoV-2 nucleic acid detection is required to be carried out in a second-level laboratory and a third-level protection, which is difficult for general grassroots hospitals, which also limits the expansion of detection capabilities; It will also affect the stability of viral nucleic acid, especially RNA, and affect the results
[0004] In the routine virus nucleic acid detection process, nucleic acid extraction is an important part of it, which generally takes 30 minutes or even longer. In response to a large number of samples such as COVID-19 and high-time nucleic acid screening work, the nucleic acid extraction process Seriously affected the screening efficiency

Method used

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  • Reagent for Respiratory Virus Treatment and Its Application
  • Reagent for Respiratory Virus Treatment and Its Application
  • Reagent for Respiratory Virus Treatment and Its Application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0076] Example 1 The composition of ready-to-use virus sampling preservation solution

[0077] Respiratory virus treatment reagent among the present invention (hereinafter referred to as ready-to-use virus sampling preservation solution, RTU) utilizes gentle nonionic detergent TWEEN20 and Nonidet P40 (NP40) to carry out cell, virus cracking, and by using dithiothreitol DTT enhanced inhibition of related nucleases such as RNase activity.

[0078] 1×RTU Composition:

[0079]

[0080] Within the above concentration range, RTU can stably perform its function.

Embodiment 2

[0081] Example 2 Ready-to-use nucleic acid detection of virus isolation cell samples and throat swab samples preserved in RTU and compatibility of conventional nucleic acid extraction kits

[0082] In the experiment, 1×RTU was used to lyse the cultured and isolated cells of ADV (DNA virus) and RSV (RNA virus) kept in the laboratory, and compared with conventional commercial nucleic acid extraction kits, the virus fluorescence quantitative PCR reagent was used for detection and evaluation RTU ready-to-use, and evaluate the compatibility of RTU-preserved samples with conventional commercial nucleic acid extraction kits.

[0083] 1. Virus isolation cells preserved in RTU can be directly used for virus nucleic acid detection and compatible with conventional nucleic acid extraction kits

[0084] (1) ADV and RSV culture

[0085] ADV culture A549 cells are cultured in DMEM+5% FBS based on 37°C 5% CO2 incubator; after the cells are full, they are digested and inoculated into 96-well ...

Embodiment 3

[0114] Example 3 RTU-preserved samples are compatible with antigen detection experiments

[0115] In order to verify whether the RTU-preserved samples are compatible with antigen-antibody analysis, the experiment carried out antigen detection on the RTU-preserved ADV samples.

[0116] (1) Culture of ADV

[0117] A549 cells were incubated in DMEM+5%FBS at 37°C in 5%CO 2 Cultivate in an incubator; cells are digested and inoculated into 96-well plates after the cells are full; the medium is removed when the cell density is about 80%, and the ADV virus seeds prepared in DMEM+2% FBS are inoculated at 35°C 5% CO 2 Cultured in the incubator for 2 days.

[0118] (2) Sample collection

[0119] After 2 days of virus culture, after aspirating the medium:

[0120] Use 1×RTU 100μl / well to lyse the cells and collect them into a 1.5ml centrifuge tube, vortex at high speed for 10sec, and let stand for 5min for virus antigen detection. The experiment is set in 3 parallels;

[0121] Use 1×...

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Abstract

The invention relates to the field of virus detection, in particular to a respiratory virus treatment reagent and its application. The respiratory virus treatment reagent is a solution comprising the following main active components: TWEEN20, Nonidet P40 and dithiothreitol. The respiratory virus treatment reagent provided by the present invention can greatly improve the screening efficiency, reduce the risk of personnel infection in the process of sample transfer and detection operation, reduce the protection level of the detection laboratory operation process, save the cost of transfer and storage, and prevent respiratory virus epidemics such as The prevention and control of COVID‑19 is of great significance.

Description

technical field [0001] The invention relates to the field of virus detection, in particular to a respiratory virus treatment reagent and its application. Background technique [0002] Respiratory tract infection has always been the most common disease with the highest incidence rate for human beings. It is the cause of death of the elderly and children. It is also the main cause of new and sudden infectious diseases, and respiratory virus infection accounts for 70% of them. -80%, such as the outbreak of novel coronavirus pneumonia (COVID-19) that occurred in mid-December 2019. As of April 25, 2020, it has caused more than 2.6 million infections and nearly 200,000 deaths worldwide, and is still developing rapidly. bring great disaster to mankind. In the prevention and control of the epidemic caused by respiratory viruses, early detection, early isolation, and early treatment are the most effective means of prevention and control. Therefore, safe and rapid viral nucleic acid...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N7/00C12Q1/70C12Q1/6806G01N33/569C12R1/93
CPCC12N7/00C12N2710/10311C12N2760/16011C12N2760/18011C12N2770/20011C12Q1/6806C12Q1/701G01N33/56983G01N2333/075G01N2333/11G01N2333/115G01N2333/135G01N2333/165C12Q2527/125
Inventor 刘文宽田新贵邱淑燕王长兵周荣
Owner THE FIRST AFFILIATED HOSPITAL OF GUANGZHOU MEDICAL UNIV (GUANGZHOU RESPIRATORY CENT)
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