CN114480559A - Disposable inactivated virus sampling tube - Google Patents
Disposable inactivated virus sampling tube Download PDFInfo
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- CN114480559A CN114480559A CN202210034445.6A CN202210034445A CN114480559A CN 114480559 A CN114480559 A CN 114480559A CN 202210034445 A CN202210034445 A CN 202210034445A CN 114480559 A CN114480559 A CN 114480559A
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- tube
- mtm
- virus
- nucleic acid
- sampling tube
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/02—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving viable microorganisms
- C12Q1/24—Methods of sampling, or inoculating or spreading a sample; Methods of physically isolating an intact microorganisms
Abstract
The invention relates to the field of microbial technology detection, in particular to a disposable inactivated virus sampling tube, wherein the components of a preservation solution comprise 150-250g/L guanidine isothiocyanate, 80-160g/L guanidine hydrochloride, 2-8g/L lead stearate, 0.1-0.6g/L Tris (2-carboxyethyl) phosphine, 50-120mL/L Tris-HCl, 5-15mL/L disodium ethylenediamine tetraacetic acid, 100-130mL/L organic alcohol and 1-5mL/L concentrated solution of an antifoaming agent A. Compared with the prior art, the invention has simple formula and convenient operation; the nucleic acid can be well preserved at extremely low temperature by adding the anti-freezing agent virus tube MTM-PRO; although the preservation effect of MTM in the virus tube without the cryoprotectant is slightly inferior to that of MTM-PRO, the nucleic acid can still be preserved intact.
Description
Technical Field
The invention relates to the field of microbial technology detection, in particular to a disposable inactivated virus sampling tube.
Background
The rapid and accurate etiology diagnosis is one of the important means for controlling the spread of epidemic situation. At present, a virus sampling tube is a complete set of sampling and conveying centrifuge tube for sampling and conveying viruses such as influenza virus, hand-foot-and-mouth virus, new coronavirus and the like in a microorganism sampling and conveying tube, and is also called a specimen conveying tube. Because the nucleic acid detection is influenced by the disease course, the specimen collection, the detection process, the detection reagent and other factors, in order to improve the positive detection rate and reduce the occurrence of false negative phenomenon, the collection and the transportation of the specimen are regulated, and the accuracy of the subsequent nucleic acid detection result is ensured. The collection, storage and transportation of nucleic acid detection samples also put higher requirements on clinic. Particularly in severe cold winter, the sample preservation solution is very easy to freeze in the transportation process, and further expands in volume, so that the sample collection tube is broken, and leakage of dangerous pathogens is very easy to occur.
The Chinese patent with publication number CN113549672A discloses an antifreezing preservation solution consisting of an acid-base indicator, pH buffer salt, a protein denaturant, an antifreezing agent, antibiotics and a solvent, and a sampling tube can be preserved for a long time at the temperature of minus 30 ℃. Considering that the formula is complex, the method aims to improve the disposable inactivated virus sampling tube which has the advantages of intact nucleic acid preservation at lower temperature, simple formula and convenient operation.
Disclosure of Invention
The invention aims to solve the defects of the prior art and provide a disposable inactivated virus sampling tube which can be used for completely storing virus nucleic acid at extremely low temperature, has a simple formula and is more convenient to operate.
In order to achieve the purpose, a disposable inactivated virus sampling tube is designed, and the components of the preservation solution are 150-250g/L guanidinium isothiocyanate, 80-160g/L guanidinium hydrochloride, 2-8g/L lead stearate, 0.1-0.6g/L Tris (2-carboxyethyl) phosphine, 50-120mL/L Tris-HCl, 5-15mL/L disodium ethylenediaminetetraacetate, 100-130mL/L organic alcohol and 1-5mL/L concentrated solution of an antifoaming agent A.
Preferably, the preservation solution further comprises an antifreeze agent and an indicator.
Preferably, the antifreeze agent is ethylene glycol and the indicator agent is phenol red.
Compared with the prior art, the invention has the advantages that:
1. the formula is simple, and the operation is convenient.
2, the MTM-PRO virus tube can still well preserve nucleic acid at extremely low temperature; MTM virus tubes have no cryoprotectant, and although the preservation effect is slightly inferior to MTM-PRO, the nucleic acid can still be preserved perfectly.
Detailed Description
The present invention is further described below in conjunction with specific embodiments, the structure and principles of which will be apparent to those skilled in the art. It should be understood that the specific embodiments described herein are merely illustrative of the invention and are not intended to limit the invention.
The swab sampling head is made of artificial fiber, synthetic fiber or flocking fiber and the like, and has milky to yellowish appearance, no stain, no rough edge and no burr; the swab liquid absorption amount is more than or equal to 0.1mL (the absorption time is 30-60 seconds).
And selecting a proper sampling tube according to the conditions of the quantity, the temperature and the like of the collected sample in the detection.
Table-sampling tube specification
The using method comprises the following steps:
1. and taking out the sample tube, and before sampling, marking related sample information or sticking a bar code label on the label of the sample tube.
2. And after sampling, quickly putting the sample into a sampling tube and screwing a tube cover.
3. The freshly collected sample should be sent to the laboratory as soon as possible, and if the sample is used for virus nucleic acid detection, the purified nucleic acid should be extracted as soon as possible; if the long-term storage is needed, the product should be stored at-40 to-70 ℃.
Example 1:
the preparation method comprises the following steps of preparing 220g/L guanidine isothiocyanate, 112g/L guanidine hydrochloride, 6.3g/L lead stearate, 0.3g/L Tris (2-carboxyethyl) phosphine, 66mL/L Tris-HCl, 8.5mL/L disodium ethylenediamine tetraacetic acid, 120mL/L organic alcohol, 2.5mL/L concentrated solution of defoamer A, ethylene glycol and phenol red to obtain a preservation solution MTM-PRO.
Throat sampling is carried out on healthy people by using a throat swab, the throat swab is stored in an MTM-PRO virus tube, and then a freezing test and a DNA and RNA co-extraction test are carried out on the MTM-PRO virus tube.
Example 2:
the following concentration is prepared, 220g/L guanidinium isothiocyanate, 112g/L guanidinium hydrochloride, 6.3g/L lead stearate, 0.3g/L Tris (2-carboxyethyl) phosphine, 66mL/L Tris-HCl, 8.5mL/L disodium ethylenediamine tetraacetic acid, 120mL/L organic alcohol and 2.5mL/L concentrated solution of defoamer A are obtained, and the preservation solution MTM is obtained.
Throat sampling was performed on healthy persons using throat swabs, which were stored in MTM virus tubes, and then a freezing test and a DNA/RNA co-extraction test were performed on the MTM virus tubes in which the swabs were stored.
Comparative example 1:
the throat of a healthy person was sampled with a throat swab containing 0.9% sodium chloride at the following concentration, the throat swab was stored in a control virus tube, and then the control virus tube was subjected to a freezing test and a DNA/RNA co-extraction test.
The freezing tests of examples 1-2 and comparative example 1 were carried out at 2-8 ℃, -10 ℃, -20 ℃, -30 ℃, -40 ℃, -80 ℃ respectively, and the freezing conditions of the virus tube at different temperatures and the preservation conditions of DNA/RNA in the pharyngeal swab at different temperatures were observed.
The freezing test is carried out in a conventional household refrigerator and an ultra-low temperature refrigerator;
the concentrated solution of the defoaming agent A is Antifoam A concentrate-100G;
DNA/RNA extraction kit the kit for semer fly was used: PureLinkTM Viral RNA/DNA Mini Kit。
The concentration and purity of the nucleic acids of examples 1-2 and comparative example 1 were measured by an ultraviolet spectrophotometer, in which the purity of the nucleic acid was the ratio of A260 to A280, A260 represents the absorbance of the nucleic acid at a wavelength of 260nm, and A280 represents the absorbance of the nucleic acid at a wavelength of 280 nm. The test results are as follows:
in Table II, it is observed whether or not the preservation solution of examples 1-2 and comparative example 1 was frozen after being preserved at temperatures of 2 to 8 ℃, -10 ℃, -20 ℃, -30 ℃, -40 ℃, -80 ℃ for 48 hours. The results show that the MTM-PRO preservative fluid can keep liquid state without freezing at the temperature of minus 30 ℃, and the MTM and the physiological saline virus tube without the preservative fluid can freeze at the temperature of minus 10 ℃.
Watch two
In Table III, the samples of examples 1-2 and comparative example 1 were stored at 2 to 8 ℃, -10 ℃, -20 ℃, -30 ℃, -40 ℃, -80 ℃ for 48 hours, and then the stored tubes were examined for frost cracking. The result shows that the virus tube of the product can not be frozen and cracked under the condition that the preservation solution is frozen at extremely low temperature.
Watch III
The nucleic acid extraction conditions of the swab samples stored in examples 1-3 were shown in Table IV after storing the samples at 2-8 ℃, -10 ℃, -20 ℃, -30 ℃, -40 ℃ and-80 ℃ for 48 hours. The result shows that the MTM-PRO virus tube can still well preserve nucleic acid at extremely low temperature; the preservation effect of the MTM virus tube without the antifreeze is slightly worse than that of MTM-PRO; the storage tube of physiological saline can detect nucleic acid only at 2-8 deg.C, and no nucleic acid can be detected at a lower temperature.
Watch four
The MTM-PRO virus tube preservation solution can be kept in a liquid state at the temperature of minus 30 ℃, and can still effectively preserve nucleic acid even under the condition that the extremely low-temperature virus tube preservation solution is frozen. Although the preservation solution for MTM virus tubes freezes at low temperature below zero, nucleic acids can be effectively preserved.
The above description is only for the purpose of illustrating the preferred embodiments of the present invention and is not to be construed as limiting the invention, and any modifications, equivalents, improvements and the like that fall within the spirit and principle of the present invention are intended to be included therein.
Claims (3)
1. The disposable inactivated virus sampling tube is characterized in that the components of the preservation solution are 150-250g/L guanidinium isothiocyanate, 80-160g/L guanidinium hydrochloride, 2-8g/L lead stearate, 0.1-0.6g/L tri (2-carboxyethyl) phosphine, 50-120mL/L Tris-HCl, 5-15mL/L disodium ethylenediaminetetraacetate, 100-130mL/L organic alcohol and 1-5mL/L defoamer A concentrated solution.
2. The single-use inactivated virus sampling tube according to claim 1, further comprising an anti-freezing agent, an indicator.
3. The single-use inactivated virus sampling tube according to claim 1 or 2, wherein the cryoprotectant is ethylene glycol and the indicator is phenol red.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202210034445.6A CN114480559A (en) | 2022-01-13 | 2022-01-13 | Disposable inactivated virus sampling tube |
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| Application Number | Priority Date | Filing Date | Title |
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| CN202210034445.6A CN114480559A (en) | 2022-01-13 | 2022-01-13 | Disposable inactivated virus sampling tube |
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| CN202210034445.6A Pending CN114480559A (en) | 2022-01-13 | 2022-01-13 | Disposable inactivated virus sampling tube |
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Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111424031A (en) * | 2020-03-20 | 2020-07-17 | 浙江安诺优达生物科技有限公司 | Composite buffer solution and kit suitable for coronavirus extraction, purification and storage |
| CN112410133A (en) * | 2020-11-20 | 2021-02-26 | 山东消博士消毒科技股份有限公司 | Environment-friendly laundry detergent and preparation method thereof |
| US20210079450A1 (en) * | 2019-09-17 | 2021-03-18 | Longhorn Vaccines And Diagnostics, Llc | Multipurpose Compositions for Collecting and Transporting Biological Material |
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- 2022-01-13 CN CN202210034445.6A patent/CN114480559A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20210079450A1 (en) * | 2019-09-17 | 2021-03-18 | Longhorn Vaccines And Diagnostics, Llc | Multipurpose Compositions for Collecting and Transporting Biological Material |
| CN111424031A (en) * | 2020-03-20 | 2020-07-17 | 浙江安诺优达生物科技有限公司 | Composite buffer solution and kit suitable for coronavirus extraction, purification and storage |
| CN112410133A (en) * | 2020-11-20 | 2021-02-26 | 山东消博士消毒科技股份有限公司 | Environment-friendly laundry detergent and preparation method thereof |
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