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Blood virus RAN protective agent and blood sampling tube

A protective agent and virus technology, which is applied in the field of molecular biology detection, can solve the problems of contaminating virus RNA, easy damage of cell membrane, and easy degradation and disappearance, so as to prevent degradation and contamination, ensure accuracy and effectiveness, prevent degradation and pollution effect

Active Publication Date: 2019-04-26
NINGBO AJCORE BIOSCIENCES INC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the content of viral RNA in the blood is small, while the content of RNase in the blood is high, and the viral RNA is easily degraded and disappears without protection; moreover, the blood is easy to coagulate during the storage and transportation of the isolated blood, and the cells in the blood The cell membrane is also easy to damage and contaminate viral RNA, so it is necessary to develop an effective and feasible protective agent that prolongs the shelf life of viral RNA

Method used

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  • Blood virus RAN protective agent and blood sampling tube
  • Blood virus RAN protective agent and blood sampling tube
  • Blood virus RAN protective agent and blood sampling tube

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] The preparation of embodiment 1 reagent

[0033] According to different anticoagulants and membrane protective agents and different prescription contents, the formulations were divided into three groups. For the sake of simplifying factors, the following experimental formulations were used: glycine 150mg / ml, aprotinin 1mg / ml, wortmannin 1mg / ml, glutathione 20mg / ml, sodium thiocyanate 100mg / ml. Prepare 3 groups of blood virus RNA protection agents. The specific formula is shown in the table below. During the preparation, the various reagents required are dissolved in DEPC water.

[0034] Table 1 Formula based on EDTA-3K anticoagulant

[0035]

[0036] Table 2 Formulas based on EDDHA–Na anticoagulant

[0037]

[0038] Table 3 Formulas based on EDTA-3K and EDDHA–Na anticoagulants

[0039]

[0040] The above protective agent is filled into a glass blood collection tube according to 0.02ml per milliliter of blood collection volume, and vacuumized to make a negati...

Embodiment 2

[0041] Example 2 Sample hemolysis performance comparison

[0042] Recruit 3 male volunteers aged 22-25 (after physical examination: normal blood routine indicators, no physical examination tumor indications, no inflammation indications), blood collection 100ml (formulation 1-3 blood collection tubes, Kangjieheparin lithium blood collection tubes, Streck noninvasive vacuum blood collection tube, blank glass blood collection tube). Store in a thermostat at room temperature at 23°C until the experiment begins.

[0043] Perform the following tests on each sample on days 0 (within 2 hours after blood collection), 2, 5, 10, 14, and 21:

[0044] For hemolysis, check the 414nm OD value.

[0045] The result is as follows:

[0046] Table 2 Hemolysis

[0047]

[0048] The results showed that, except Kangjie Lithium heparin blood collection tube and blank glass blood collection tube, the hemolysis status of other blood collection tubes was better on the 21st day, especially on the 15...

Embodiment 3

[0049] Embodiment 3: virus RNA preservation effect experiment

[0050] Source of experimental samples: 2016 HCV-positive patients in the First Affiliated Hospital of Zhejiang Medical University;

[0051] Sample collection: Collect blood samples with formula 1-3 blood collection tubes, Streck non-invasive vacuum blood collection tubes, and blank glass blood collection tubes, 5 of each blood collection tube, 1ml of blood for each tube, and keep them in a room temperature thermostat at 23 degrees Celsius until the start experiment.

[0052] Take the 0th, 2nd, 5th, 10th, and 14th days for virus detection on each sample;

[0053] HCV viral RNA detection:

[0054] Nucleic acid was extracted using Roche nucleic acid extraction column, amplified and detected with HCV fluorescent quantitative PCR detection kit of Shenzhen Piji Company and Opticon fluorescent quantitative PCR detector of American Research Company; in order to avoid batch-to-batch differences in nucleic acid quantifica...

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PUM

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Abstract

The invention provides a blood virus RAN protective agent and a blood sampling tube. The blood virus RAN protective agent is prepared by dissolving the following components in DEPC (diethyl pyrocarbonate) solution 100-500mg / ml glycine, 1-5mg / ml aprotinin, 1-5mg / ml wortmannin, 10-40mg / ml glutathione, 100-500mg / ml sodium thiocyanate, 1-3mg / ml anticoagulant and 5-20mg / ml membrane protective agent. The blood virus RAN protective agent provided by the invention is capable of supplying long-term protection to virus RNA in blood under room temperature, preventing degradation and pollution of virus RNA and effectively guaranteeing accuracy and validity of blood sample.

Description

technical field [0001] The application belongs to the field of molecular biology detection and the field of medical equipment, and in particular relates to a blood virus RAN protective agent and a blood collection tube. Background technique [0002] Viruses consist of a nucleic acid molecule and protein or only protein (such as prions). Viruses are very small in size and extremely simple in structure, but they are highly parasitic and completely dependent on the energy and metabolic systems of host cells. When it comes into contact with the host cell, it takes off the protein coat, and its nucleic acid (gene) invades the host cell, and by means of the latter's replication system, it replicates new viruses according to the instructions of the viral gene, and finally causes the death of the host cell. There are very few DNA viruses in nature, and they are basically RNA viruses, such as HIV, tobacco mosaic virus, SARS virus, MERS virus, Ebola virus (EBV), Spanish flu virus, H1...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/10C12M1/24
CPCC12M23/06C12N15/10
Inventor 周杰锋王德明
Owner NINGBO AJCORE BIOSCIENCES INC
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