CN112779316A - Inactivated virus preservation solution and preparation method thereof - Google Patents
Inactivated virus preservation solution and preparation method thereof Download PDFInfo
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- 241000700605 Viruses Species 0.000 title claims abstract description 76
- 239000003761 preservation solution Substances 0.000 title claims abstract description 44
- 238000002360 preparation method Methods 0.000 title claims abstract description 8
- 229940080258 tetrasodium iminodisuccinate Drugs 0.000 claims abstract description 24
- GYBINGQBXROMRS-UHFFFAOYSA-J tetrasodium;2-(1,2-dicarboxylatoethylamino)butanedioate Chemical compound [Na+].[Na+].[Na+].[Na+].[O-]C(=O)CC(C([O-])=O)NC(C([O-])=O)CC([O-])=O GYBINGQBXROMRS-UHFFFAOYSA-J 0.000 claims abstract description 24
- VHJLVAABSRFDPM-QWWZWVQMSA-N dithiothreitol Chemical compound SC[C@@H](O)[C@H](O)CS VHJLVAABSRFDPM-QWWZWVQMSA-N 0.000 claims abstract description 14
- QKNYBSVHEMOAJP-UHFFFAOYSA-N 2-amino-2-(hydroxymethyl)propane-1,3-diol;hydron;chloride Chemical compound Cl.OCC(N)(CO)CO QKNYBSVHEMOAJP-UHFFFAOYSA-N 0.000 claims abstract description 11
- YQOKLYTXVFAUCW-UHFFFAOYSA-N guanidine;isothiocyanic acid Chemical compound N=C=S.NC(N)=N YQOKLYTXVFAUCW-UHFFFAOYSA-N 0.000 claims abstract description 11
- 239000003945 anionic surfactant Substances 0.000 claims abstract description 8
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- 229960000686 benzalkonium chloride Drugs 0.000 claims description 9
- CADWTSSKOVRVJC-UHFFFAOYSA-N benzyl(dimethyl)azanium;chloride Chemical group [Cl-].C[NH+](C)CC1=CC=CC=C1 CADWTSSKOVRVJC-UHFFFAOYSA-N 0.000 claims description 9
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- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 8
- 239000000243 solution Substances 0.000 claims description 7
- MCHZKGNHFPNZDP-UHFFFAOYSA-N 2-aminoethane-1,1,1-triol;hydrochloride Chemical compound Cl.NCC(O)(O)O MCHZKGNHFPNZDP-UHFFFAOYSA-N 0.000 claims description 5
- 229940083575 sodium dodecyl sulfate Drugs 0.000 claims description 5
- 238000000034 method Methods 0.000 claims description 3
- 239000012528 membrane Substances 0.000 claims description 2
- RWZXIUKKMTXBQY-UHFFFAOYSA-M sodium dodecanoylazanide Chemical compound C(CCCCCCCCCCC)(=O)[NH-].[Na+] RWZXIUKKMTXBQY-UHFFFAOYSA-M 0.000 claims description 2
- 238000003756 stirring Methods 0.000 claims description 2
- 235000019333 sodium laurylsulphate Nutrition 0.000 claims 4
- 102000039446 nucleic acids Human genes 0.000 abstract description 28
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- 150000007523 nucleic acids Chemical class 0.000 abstract description 28
- 238000001514 detection method Methods 0.000 abstract description 19
- 230000002779 inactivation Effects 0.000 abstract description 13
- 208000015181 infectious disease Diseases 0.000 abstract description 8
- 238000000605 extraction Methods 0.000 abstract description 7
- 230000003321 amplification Effects 0.000 abstract description 5
- 230000000415 inactivating effect Effects 0.000 abstract description 5
- 238000003199 nucleic acid amplification method Methods 0.000 abstract description 5
- 239000012472 biological sample Substances 0.000 abstract description 4
- 230000005540 biological transmission Effects 0.000 abstract description 2
- 230000000694 effects Effects 0.000 description 15
- 241001112090 Pseudovirus Species 0.000 description 9
- 239000000523 sample Substances 0.000 description 9
- 108010043121 Green Fluorescent Proteins Proteins 0.000 description 8
- 102000004144 Green Fluorescent Proteins Human genes 0.000 description 8
- 239000005090 green fluorescent protein Substances 0.000 description 8
- 150000002357 guanidines Chemical class 0.000 description 6
- 210000004027 cell Anatomy 0.000 description 5
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- 238000003860 storage Methods 0.000 description 4
- DPXJVFZANSGRMM-UHFFFAOYSA-N acetic acid;2,3,4,5,6-pentahydroxyhexanal;sodium Chemical compound [Na].CC(O)=O.OCC(O)C(O)C(O)C(O)C=O DPXJVFZANSGRMM-UHFFFAOYSA-N 0.000 description 3
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- DAJSVUQLFFJUSX-UHFFFAOYSA-M sodium;dodecane-1-sulfonate Chemical compound [Na+].CCCCCCCCCCCCS([O-])(=O)=O DAJSVUQLFFJUSX-UHFFFAOYSA-M 0.000 description 3
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- 108090000623 proteins and genes Proteins 0.000 description 2
- 239000012264 purified product Substances 0.000 description 2
- 238000003753 real-time PCR Methods 0.000 description 2
- 239000004094 surface-active agent Substances 0.000 description 2
- 238000012795 verification Methods 0.000 description 2
- 210000003501 vero cell Anatomy 0.000 description 2
- 230000003612 virological effect Effects 0.000 description 2
- 238000005406 washing Methods 0.000 description 2
- 238000005303 weighing Methods 0.000 description 2
- KCXVZYZYPLLWCC-UHFFFAOYSA-N EDTA Chemical compound OC(=O)CN(CC(O)=O)CCN(CC(O)=O)CC(O)=O KCXVZYZYPLLWCC-UHFFFAOYSA-N 0.000 description 1
- 108010067770 Endopeptidase K Proteins 0.000 description 1
- 101710163270 Nuclease Proteins 0.000 description 1
- 208000032370 Secondary transmission Diseases 0.000 description 1
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 1
- BFNBIHQBYMNNAN-UHFFFAOYSA-N ammonium sulfate Chemical compound N.N.OS(O)(=O)=O BFNBIHQBYMNNAN-UHFFFAOYSA-N 0.000 description 1
- 229910052921 ammonium sulfate Inorganic materials 0.000 description 1
- 235000011130 ammonium sulphate Nutrition 0.000 description 1
- 239000000427 antigen Substances 0.000 description 1
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- 239000008280 blood Substances 0.000 description 1
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- 239000003153 chemical reaction reagent Substances 0.000 description 1
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- 230000002147 killing effect Effects 0.000 description 1
- 238000002156 mixing Methods 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 239000003755 preservative agent Substances 0.000 description 1
- 230000002335 preservative effect Effects 0.000 description 1
- 230000000644 propagated effect Effects 0.000 description 1
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- 238000003757 reverse transcription PCR Methods 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 238000012163 sequencing technique Methods 0.000 description 1
- 239000001509 sodium citrate Substances 0.000 description 1
- NLJMYIDDQXHKNR-UHFFFAOYSA-K sodium citrate Chemical compound O.O.[Na+].[Na+].[Na+].[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O NLJMYIDDQXHKNR-UHFFFAOYSA-K 0.000 description 1
- 230000007480 spreading Effects 0.000 description 1
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- 230000001954 sterilising effect Effects 0.000 description 1
- 238000004659 sterilization and disinfection Methods 0.000 description 1
- QAOWNCQODCNURD-UHFFFAOYSA-N sulfuric acid Substances OS(O)(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-N 0.000 description 1
- 231100000331 toxic Toxicity 0.000 description 1
- 230000002588 toxic effect Effects 0.000 description 1
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- 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
- C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
- C12Q1/6806—Preparing nucleic acids for analysis, e.g. for polymerase chain reaction [PCR] assay
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Abstract
The invention relates to an inactivated virus preservation solution and a preparation method thereof, wherein the inactivated virus preservation solution comprises guanidine isothiocyanate, tris hydrochloride, tetrasodium iminodisuccinate, an anionic surfactant, a cationic surfactant and dithiothreitol. The inactivated virus preservation solution provided by the invention not only effectively ensures the integrity of virus nucleic acid, but also adds components for inactivating the virus, thereby ensuring the inactivation performance of the virus, enabling the collected biological sample to be applied to nucleic acid detection, reducing the risk of virus transmission infection, and not influencing subsequent nucleic acid extraction and amplification detection.
Description
Technical Field
The invention relates to an inactivated virus preservation solution and a preparation method thereof, belonging to the technical field of in vitro diagnosis.
Background
Viruses are non-cellular in life forms, must be parasitically propagated within living cells, are simple in structure, and generally consist of nucleic acids (DNA or RNA) and a protein coat encapsulating the nucleic acids. Most viruses are pathogenic microorganisms that cause host morbidity and can be transmitted in organisms to become infectious. At present, the in vitro diagnosis can be carried out by collecting samples such as throat swabs, nose swabs and blood of patients to detect whether a certain virus is infected, the common detection method is to carry out antigen antibody detection or nucleic acid detection on the samples of the patients, but the virus generally leaves host cells and can be inactivated quickly, the integrity of the virus can be destroyed, the virus shell is disintegrated, and the nucleic acid is degraded, so that in order to ensure the accuracy of the in vitro diagnosis, the samples of the virus are transported and stored by adopting specific instruments or a storage solution.
At present, the main virus preservation solution in the market is generally normal saline, PBS buffer solution or UTM transport medium, and the virus preservation solution can be applied to nucleic acid detection, but because no inactivation component is added or the inactivation effect is poor, the collected biological sample has the risk of secondary transmission infection.
Currently, most of the commercially available inactivated virus preservation solutions are inactivated by adopting high-concentration guanidine salt, although the high-concentration guanidine salt has a good virus inactivation effect, the high-concentration guanidine salt is easy to crystallize and separate out at low temperature (for example, collected samples are transported in northern low-temperature areas in winter), the virus inactivation effect can be reduced after separation, and the risk of spreading infection in the transportation process is increased; on the other hand, the difficulty of washing and removing the high-concentration guanidine salt in the nucleic acid extraction process is increased, guanidine salt residues in the nucleic acid are easily caused, the guanidine salt residues can inhibit the amplification of the nucleic acid, and further the false negative sample ratio is increased due to the fact that detection is missed.
Disclosure of Invention
Technical problem to be solved
In order to solve the above problems in the prior art, the present invention provides an inactivated virus preservation solution, which is added with a component for inactivating viruses on the basis of ensuring the integrity of viral nucleic acids, so that the collected biological sample can be applied to nucleic acid detection, and the risk of virus transmission infection is reduced.
(II) technical scheme
In order to achieve the purpose, the invention adopts the main technical scheme that:
an inactivated virus preservation solution comprises guanidinium isothiocyanate, tris hydrochloride, tetrasodium iminodisuccinate, an anionic surfactant, a cationic surfactant and dithiothreitol.
In the inactivated virus preservation solution as described above, preferably, the anionic surfactant is sodium dodecyl sulfate or sodium lauroyl amide, and the cationic surfactant is benzalkonium chloride.
Further, the anionic surfactant is preferably sodium dodecyl sulfate.
The inactivated virus preservation solution as described above preferably comprises the following components in final concentration: the guanidine isothiocyanate is 0.1-1 mol/L, the trihydroxymethyl aminomethane hydrochloride is 10-300 mmol/L, the tetrasodium iminodisuccinate is 1-50 mmol/L, and the weight volume ratio of the guanidine isothiocyanate to the tetrasodium iminodisuccinate is 0.1-2%, the weight volume ratio of the tetrasodium iminodisuccinate is 0.01-0.2%, and the weight volume ratio of the tetrasodium iminodisuccinate is 0.01-0.2%.
The inactivated virus preservation solution as described above preferably comprises the following components in final concentration: the guanidine isothiocyanate is 0.1mol/L, the tris hydrochloride is 100mmol/L, the tetrasodium iminodisuccinate is 30mmol/L, and the weight volume ratio of the tetrasodium iminodisuccinate is 0.5%, the cationic surfactant is 0.05%, and the dithiothreitol is 0.1%.
The inactivated virus preservation solution preferably has a pH of 5.0 to 8.0, more preferably 6.0.
The preparation method of the inactivated virus preservation solution comprises the following steps: weighing guanidinium isothiocyanate, trihydroxymethyl aminomethane hydrochloride, tetrasodium iminodisuccinate, sodium dodecyl sulfate, benzalkonium chloride and dithiothreitol, adding 800mL of purified water, stirring for dissolving, adjusting the pH value to 5.0-8.0 by NaOH, adding the purified water to a constant volume of 1L, so that the concentration of the guanidinium isothiocyanate is 0.1-1 mol/L, 10-300 mmol/L of the trihydroxymethyl aminomethane hydrochloride, 1-50 mmol/L of tetrasodium iminodisuccinate, 0.1-2% by weight-volume ratio (w/v, g/mL) of the sodium dodecyl sulfate, 0.01-0.2% by weight-volume ratio (w/v, g/mL) of the benzalkonium chloride, 0.01-0.2% by weight-volume ratio (w/v, g/mL) of the dithiothreitol, and after preparation, sterilizing the solution by passing through a 0.25 mu m filter membrane.
(III) advantageous effects
The invention has the beneficial effects that:
compared with the prior art, the inactivated virus preservation solution provided by the invention not only effectively ensures the integrity of virus nucleic acid, but also adds a component for inactivating the virus, so that the collected biological sample can be applied to nucleic acid detection, the risk of virus propagation infection is reduced, the degradation of the nucleic acid can be effectively avoided, and the accuracy of the detection result is ensured.
The inactivated virus preservation solution provided by the invention ensures the inactivation performance of the virus, provides nucleic acid with quality assurance for subsequent fluorescence detection or sequencing, does not influence the subsequent nucleic acid extraction and amplification detection results, and has good stability. Meanwhile, the biodegradable tetrasodium iminodisuccinate is used, so that the environmental pollution is reduced.
Drawings
FIG. 1 shows the result of detecting the expression of green fluorescent protein under a fluorescence microscope;
FIG. 2 shows the result of detecting the expression of green fluorescent protein in the control group under a fluorescent microscope;
FIG. 3 is 103The effect of the copies/mL pseudovirus is preserved at room temperature;
FIG. 4 is 104The effect of the copies/mL pseudovirus is preserved at room temperature;
FIG. 5 is 105The effect of copies/mL pseudovirus was preserved at room temperature.
Detailed Description
For the purpose of better explaining the present invention and to facilitate understanding, the present invention will be described in detail by way of specific embodiments with reference to the accompanying drawings.
On the basis of a large number of experimental researches, the invention discovers that guanidine isothiocyanate can denature protein and has the effect of inactivating viruses, but the dosage is not easy to be too much, so that the solution is unstable and easy to crystallize due to too much dosage, and the effect of inactivating viruses cannot be achieved due to too little dosage, so that the optimal concentration is 0.1-1 mol/L. The adopted tetrasodium iminodisuccinate has the advantages of inhibiting nuclease activity, preventing nucleic acid degradation, being biodegradable by itself, having no toxic action and reducing environmental pollution. The anion surfactant and the protein are denatured, which is beneficial to the disintegration of virus shells, the cation surfactant is used for sterilization and disinfection and killing pathogenic microorganisms, and the dithiothreitol is used for protecting RNA. According to the invention, guanidine salt with lower concentration is adopted, so that nucleic acid is easy to remove in the washing process, various surfactants with inactivation performance are added, and the combination of various inactivation reagents is utilized, so that the concentration of guanidine salt is reduced, the inactivation performance of virus is ensured, and the subsequent nucleic acid extraction and amplification detection are not influenced.
Example 1
Preparing an inactivated virus preservation solution: 59.08g of guanidinium isothiocyanate, 15.76g of tris (hydroxymethyl) aminomethane hydrochloride, 6.74g of tetrasodium iminodisuccinate, 5g of sodium dodecyl sulfonate, 0.5g of benzalkonium chloride and 1g of dithiothreitol are weighed, 800mL of purified water is added, the pH value is adjusted to 6.0 by NaOH, the volume is fixed to 1L by adding the purified water, so that the final concentration of the guanidinium isothiocyanate is 0.1mol/L, the tris (hydroxymethyl) aminomethane hydrochloride is 100mmol/L, the tetrasodium iminodisuccinate is 30mmol/L, the weight-to-volume ratio of the sodium dodecyl sulfonate is 0.50% (w/v), the weight-to-volume ratio of the benzalkonium chloride is 0.05% (w/v), and the weight-to-volume ratio of the dithiothreitol is 0.10% (w/v). After preparation, the solution was sterilized through a 0.25 μm filter.
1. Verification of virus inactivation performance
The experimental group adopts lentivirus of high-brightness green fluorescent protein and the prepared virus preservation solution according to the volume ratio of 1: 1 Mixed storage to a final virus titer of 108And TU/mL, mixing for 10 minutes, then taking the mixture to infect Vero cells, culturing the infected cells for 3 days, and detecting the expression of green fluorescent protein under a fluorescence microscope to verify the activity of the virus. Control group: lentivirus using high brightness green fluorescent protein (Virus Titer 10)8TU/mL) directly infect Vero cells, and the infected cells are cultured for 3 days, and then the expression of green fluorescent protein is detected under a fluorescence microscope to verify the activity of the virus. The results are shown in fig. 1, which is the result of detecting the expression of green fluorescent protein under a fluorescence microscope for the experimental group, and it can be seen that under the fluorescence microscope, there is no green color, which indicates that the virus has been inactivated by the virus preservation solution and has no ability to infect cells. FIG. 2 shows that the control group detected the expression of green fluorescent protein under the fluorescence microscope and produced obvious green fluorescence under the fluorescence microscope, which indicates that the virus has very high green fluorescenceStrong infection capacity.
The experimental result shows that the lentivirus is completely inactivated after being preserved in the virus preservation solution for 10 minutes, which shows that the virus preservation solution of the invention has better virus inactivation effect.
Example 2 viral nucleic acid preservation Effect and nucleic acid detection verification
1, sample treatment: the virus preservation solution prepared in example 1 was sampled from each experimental batch and labeled Cellpro 3 tube (2 mL/tube) and the virus preservation solution prepared according to the prior art (wherein, the final concentration of sodium citrate is 25mmol/L, the final concentration of proteinase K is 200. mu.g/mL, the final concentration of concentrated sulfuric acid is 2.5% (V/V), the final concentration of ethylenediaminetetraacetic acid is 30mmol/L, and the final concentration of ammonium sulfate is 60% (W/V)) was labeled ComPA 3 tube (2 mL/tube), and a gradient diluted RNA pseudovirus sample (King Zhi Biotech Co., Ltd.) was added so that the concentration was 10 in order3copies/mL、104copies/mL、105And (3) preserving the copies/mL by placing the preservation solution added with the pseudovirus sample at room temperature (20-25 ℃, humidity is 50% -60%).
2, extracting virus RNA: the samples stored at room temperature are respectively taken out after being stored for 1, 2, 3, 5 and 7 days for RNA extraction and detection. 200 mu L of each preservative fluid sample is taken, and nucleic acid extraction is carried out according to the flow of a virus nucleic acid extraction kit (Cat. RC311-C1, Nanjing Novodka Biotech Co., Ltd.). The final pseudoviral RNA purified product eluted in a volume of 50. mu.L.
3RT-PCR fluorescent quantitative detection: according to the experimental flow of the invitrogen reverse transcription kit, the total reaction volume is 21 mu L, and 7 mu L of pseudovirus RNA purified product is added. After reverse transcription, according to the experimental flow of a fluorescent quantitative detection kit, the total reaction volume is 20 mu L, 5 mu L of reverse transcription product is added, an ABI-7500Fast fluorescent quantitative PCR instrument is used, and a reference kit instruction is set according to the program.
4 fluorescent quantitative PCR detection data result
The results of the detection data of the Cellpro and the CompA preservation solutions added with the samples with three concentrations are shown in fig. 3, fig. 4 and fig. 5 when the Cellpro and the CompA preservation solutions are preserved for 1 day, 2 days, 3 days, 5 days and 7 days at room temperature, and as can be seen from the graphs, the CT value of the virus preservation solution is lower as can be seen from the comparison of the nucleic acid amplification CT values of the pseudoviruses with the same concentration in the two virus preservation solutions, which indicates that the virus preservation solution has better virus preservation effect and higher concentration of extracted virus nucleic acid.
In addition, the virus sample storage, virus RNA extraction and RT-PCR complete process are carried out with the commercially available virus storage solution according to the above operation, the detected sample is RNA pseudovirus samples with different concentration gradients, and the RNA pseudovirus samples are respectively stored for 1 day, 2 days, 3 days, 5 days and 7 days at room temperature (20-25 ℃ and humidity of 50% -60%) and detected. The detection result shows that: the virus preservation solution provided by the invention is preserved for 7 days at room temperature, and compared with the existing products, the CT value change amplitude of the inactivated virus preservation solution provided by the invention is smaller, which shows that the inactivated virus preservation solution provided by the invention has higher nucleic acid extraction amount and better stability.
Comparative example 1
Weighing 59.08g of guanidinium isothiocyanate, 15.76g of tris (hydroxymethyl) aminomethane hydrochloride, 6.74g of tetrasodium iminodisuccinate, 5g of sodium dodecyl sulfonate and 1g of dithiothreitol, adding 800mL of purified water, adjusting the pH value to 6.0 by NaOH, adding the purified water to a constant volume of 1L, so that the concentration of the guanidinium isothiocyanate is 0.1mol/L, 100m mol/L of tris (hydroxymethyl) aminomethane hydrochloride, 30m mol/L of tetrasodium iminodisuccinate, 0.50% (w/v) of sodium dodecyl sulfate in a weight-to-volume ratio, and 0.10% (w/v) of dithiothreitol in a weight-to-volume ratio; this comparative example removed the benzalkonium chloride component and found that the inactivation time of lentivirus required up to 5 hours for complete inactivation, which resulted in the risk of infection.
Comparative example 2
In this example, the concentration of guanidine isothiocyanate was 2mol/L, and the other components and concentrations were the same as those in example 1, and when the sample was left at 4 ℃ for 24 hours (simulating outdoor low-temperature transportation environment), crystal precipitation occurred and the stability was poor, whereas when the sample was left at 4 ℃ for 24 hours or longer, for example, 6 months, no crystal precipitation was observed and the stability was good.
The above description is only a preferred embodiment of the present invention, and is not intended to limit the present invention in other forms, and any person skilled in the art can change or modify the technical content disclosed above into an equivalent embodiment with equivalent changes. However, any simple modification, equivalent change and modification of the above embodiments according to the technical essence of the present invention are within the protection scope of the technical solution of the present invention.
Claims (7)
1. An inactivated virus preservation solution is characterized by comprising guanidinium isothiocyanate, tris hydrochloride, tetrasodium iminodisuccinate, an anionic surfactant, a cationic surfactant and dithiothreitol.
2. The inactivated virus preservation solution according to claim 1, wherein the anionic surfactant is sodium dodecyl sulfate or sodium lauroyl amide, and the cationic surfactant is benzalkonium chloride.
3. The inactivated virus preservation solution according to claim 1, wherein the anionic surfactant is preferably sodium dodecyl sulfate.
4. The inactivated virus preservation solution according to claim 1, wherein the final concentration of each component in the preservation solution is: the guanidine isothiocyanate is 0.1-1 mol/L, the trihydroxymethyl aminomethane hydrochloride is 10-300 mmol/L, the tetrasodium iminodisuccinate is 1-50 mmol/L, and the weight volume ratio of the guanidine isothiocyanate to the trihydroxymethyl aminomethane hydrochloride is 0.1-2%, the weight volume ratio of the tetrasodium iminodisuccinate is 0.01-0.2%, and the weight volume ratio of the tetrasodium iminodisuccinate is 0.01-0.2%.
5. The inactivated virus preservation solution according to claim 1, wherein the final concentration of each component in the preservation solution is: the guanidine isothiocyanate is 0.1mol/L, the tris hydrochloride is 100mmol/L, the tetrasodium iminodisuccinate is 30mmol/L, and the weight volume ratio of the tetrasodium iminodisuccinate is 0.5%, the cationic surfactant is 0.05%, and the dithiothreitol is 0.1%.
6. The inactivated virus preservation solution according to claim 1, wherein the pH value of the preservation solution is 5.0 to 8.0.
7. The method for preparing the inactivated virus preservation solution according to any one of claims 1 to 6, wherein guanidine isothiocyanate, tris hydrochloride, tetrasodium iminodisuccinate, sodium dodecylsulfate, benzalkonium chloride, and dithiothreitol are weighed, 800mL of purified water is added and dissolved with stirring, the pH is adjusted to 5.0 to 8.0 with NaOH, purified water is added to a constant volume of 1L, the concentration of guanidine isothiocyanate is 0.1 to 1mol/L, tris hydrochloride is 10 to 300mmol/L, tetrasodium iminodisuccinate is 1 to 50mmol/L, sodium dodecylsulfate is 0.1 to 2% by weight/volume, benzalkonium chloride is 0.01 to 0.2% by weight/volume, and dithiothreitol is 0.01 to 0.2% by weight/volume, and after preparation, the solution is sterilized by passing through a 0.25 μm filter membrane.
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