CN113549672A - Anti-freezing virus sampling and preserving liquid and sampling tube - Google Patents
Anti-freezing virus sampling and preserving liquid and sampling tube Download PDFInfo
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Abstract
The invention relates to the technical field of microbial detection, and discloses an anti-freezing type virus sampling and storing liquid and a sampling tube, in order to solve the problem of leakage caused by rupture of a sample collecting tube due to freezing and volume expansion of the sample storing liquid in the transportation process in severe cold winter. The preservation solution mainly comprises an acid-base indicator, pH buffer salt, a protein denaturant, an antifreezing agent, antibiotics and a solvent. The preservation solution disclosed by the invention is low-temperature resistant, can be used for preserving samples in an environment at the temperature of-30 ℃ for a long time, is not frozen or cracked, is stable in virus nucleic acid, and is beneficial to preservation and transportation of samples in extremely cold areas in winter.
Description
Technical Field
The invention relates to the technical field of microbial detection, in particular to an anti-freezing virus sampling and preserving solution and a sampling tube.
Background
The rapid and accurate etiology diagnosis is one of the important means for controlling the spread of epidemic situation. By adopting a real-time fluorescent quantitative PCR (RT-PCR) method, the novel coronavirus nucleic acid can be detected in nasopharyngeal swab, sputum and other specimens of lower respiratory tract secretion, blood, excrement, urine and the like of an infected person.
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.
Disclosure of Invention
The invention aims to solve the defects in the prior art and provides an anti-freezing virus sampling and preserving solution and a sampling tube.
In order to achieve the purpose, the invention adopts the following technical scheme:
the anti-freezing type virus sampling and preserving fluid mainly comprises an acid-base indicator, pH buffer salt, a protein denaturant, an anti-freezing agent, antibiotics and a solvent;
the acid-base indicator comprises phenol red; the pH buffer salt comprises disodium hydrogen phosphate dodecahydrate and potassium dihydrogen phosphate; protein denaturants including guanidinium isothiocyanate and ethylphenylpolyethylene glycol; anti-freezing agents include glycerol and ethylene glycol; antibiotics include gentamicin; the solvent comprises purified water.
Preferably, each 30L of the preservation solution contains 0.001-0.004% of phenol red, 2.0-3.0 mm of disodium hydrogen phosphate dodecahydrate, 0.2-1.0 mm of potassium dihydrogen phosphate, 0.4-4 m/L of guanidine isothiocyanate, 0.3-1% of ethyl phenyl polyethylene glycol, 10-40% of glycerol, 10-40% of ethylene glycol, 0.3-3g of gentamicin and purified water with the constant volume of 30L.
Preferably, the pH value of the preservation solution is 7.0-8.0.
Preferably, each 30L of the preservation solution contains 0.001% of phenol red, 2.0mm of disodium hydrogen phosphate dodecahydrate, 0.2mm of monopotassium phosphate, 0.4m/L of guanidine isothiocyanate, 0.3% of ethyl phenyl polyethylene glycol, 30% of glycerol, 30% of ethylene glycol, 0.3g of gentamicin and purified water with the volume of 30L.
Preferably, each 30L of the preservation solution contains 0.003 percent of phenol red, 2.4mm of disodium hydrogen phosphate dodecahydrate, 0.4mm of monopotassium phosphate, 0.8m/L of guanidine isothiocyanate, 0.5 percent of ethyl phenyl polyethylene glycol, 40 percent of glycerin, 40 percent of ethylene glycol, 0.6g of gentamicin and purified water with the volume of 30L.
Preferably, each 30L of the preservation solution contains 0.004% of phenol red, 2.8mm of disodium hydrogen phosphate dodecahydrate, 0.8mm of potassium dihydrogen phosphate, 1.5m/L of guanidine isothiocyanate, 0.5% of ethyl phenyl polyethylene glycol, 40% of glycerol, 40% of ethylene glycol, 0.6g of gentamicin and purified water with the volume of 30L.
Preferably, each 30L of the preservation solution contains 0.004% of phenol red, 3.0mm of disodium hydrogen phosphate dodecahydrate, 1.0mm of potassium dihydrogen phosphate, 3m/L of guanidine isothiocyanate, 0.5% of ethyl phenyl polyethylene glycol, 40% of glycerol, 40% of ethylene glycol, 1.2g of gentamicin and purified water with the volume of 30L.
Preferably, the preparation method of the preservation solution comprises the following steps: mixing disodium hydrogen phosphate dodecahydrate, potassium dihydrogen phosphate, guanidine isothiocyanate and purified water uniformly, adding ethylphenyl polyethylene glycol, glycerol and ethylene glycol, mixing uniformly, adding phenol red, and mixing uniformly to obtain the preservation solution.
A sampling tube comprises a swab and a collecting tube, and further comprises the anti-freezing virus sampling and preserving fluid.
The invention has the beneficial effects that:
the preservation solution is inactivated, the virus completely loses infectivity in the preservation process, the biological safety is high, the preservation solution is suitable for preserving various virus samples, such as influenza virus, enterovirus, HIV virus, coronavirus and the like, the preservation effect is good, the DNA/RNA of the virus is not degraded in the preservation process, and the virus release rate is high; the preservation solution is low temperature resistant, can preserve samples for a long time in an environment of 30 ℃ below zero, is not frozen and cracked, has stable virus nucleic acid, and is beneficial to the preservation and transportation of samples in extremely cold areas in winter; the use is simple and convenient, microbial protein can be cracked and inactivated, secondary infection risk is effectively prevented, and the safety of transportation and detection personnel is guaranteed; the applicable sample range is wide: such as oral swabs, pharyngeal swabs, nasal swabs, anal swabs, cervical swabs, saliva, sputum, alveolar lavage, and the like, and can also be used to collect environmental samples.
Drawings
FIG. 1 is a data diagram of 0 day at a preservation temperature of 2-8 ℃ of an ADV1-10 sample in a preservation performance test of an antifreeze type virus sampling preservation solution provided by the invention, wherein the first line from top to bottom in the data diagram is ADV 2; the second thread is ADV 1; the third line is ADV 8; the fourth line is ADV 7; the fifth line is ADV 10; the sixth line is ADV 9; the seventh line is ADV 3; the eighth line is ADV 4; the ninth line is ADV 6; the tenth line is ADV 5;
FIG. 2 is a data diagram of 0 day at-10 ℃ storage temperature of an ADV1-10 sample in the antifreeze virus sampling storage fluid storage performance test, wherein the first line from top to bottom in the data diagram is ADV 7; the second thread is ADV 8; the third line is ADV 4; the fourth line is ADV 1; the fifth line is ADV 2; the sixth line is ADV 3; the seventh line is ADV 10; the eighth line is ADV 9; the ninth line is ADV 5; the tenth line is ADV 6;
FIG. 3 is a data diagram of 0 day at-20 ℃ storage temperature of ADV1-10 samples in the antifreeze virus sampling storage fluid storage performance test provided by the present invention, wherein the first line from top to bottom in the data diagram is ADV 2; the second thread is ADV 4; the third line is ADV 1; the fourth line is ADV 9; the fifth line is ADV 3; the sixth line is ADV 10; the seventh line is ADV 5; the eighth line is ADV 8; the ninth line is ADV 7; the tenth line is ADV 6;
FIG. 4 is a data diagram of 0 day at-30 ℃ storage temperature of an ADV1-10 sample in the antifreeze virus sampling storage fluid storage performance test, wherein the first line from top to bottom is ADV 8; the second thread is ADV 7; the third line is ADV 4; the fourth line is ADV 1; the fifth line is ADV 3; the sixth line is ADV 2; the seventh line is ADV 10; the eighth line is ADV 9; the ninth line is ADV 6; the tenth line is ADV 5;
FIG. 5 is a data diagram of 0 day at-40 ℃ storage temperature of ADV1-10 samples in the antifreeze virus sampling preservation solution storage performance test provided by the invention, wherein the first line from top to bottom in the data diagram is ADV 10; the second thread is ADV 1; the third line is ADV 2; the fourth line is ADV 6; the fifth line is ADV 9; the sixth line is ADV 8; the seventh line is ADV 3; the eighth line is ADV 4; the ninth line is ADV 7; the tenth line is ADV 5;
FIG. 6 is a data diagram of 0 day at-80 ℃ storage temperature of an ADV1-10 sample in the antifreeze virus sampling preservation solution storage performance test, wherein the first line from top to bottom in the data diagram is ADV 4; the second thread is ADV 3; the third line is ADV 7; the fourth line is ADV 8; the fifth line is ADV 1; the sixth line is ADV 5; the seventh line is ADV 6; the eighth line is ADV 9; the ninth line is ADV 10; the tenth line is ADV 2;
FIG. 7 is a data diagram of ADV1-10 samples stored at 2-80 deg.C for 7 days in the antifreeze virus sampling preservation solution storage performance test, the first line from top to bottom in the data diagram is ADV 3; the second thread is ADV 4; the third line is ADV 6; the fourth line is ADV 5; the fifth line is ADV 8; the sixth line is ADV 7; the seventh line is ADV 1; the eighth line is ADV 2; the ninth line is ADV 9; the tenth line is ADV 10;
FIG. 8 is a data diagram of the preservation temperature of the sample ADV1-10 at-10 ℃ for 7 days in the preservation performance test of the antifreeze virus sampling preservation solution provided by the invention, wherein the first line from top to bottom in the data diagram is ADV 7; the second thread is ADV 9; the third line is ADV 10; the fourth line is ADV 5; the fifth line is ADV 8; the sixth line is ADV 6; the seventh line is ADV 4; the eighth line is ADV 3; the ninth line is ADV 2; the tenth line is ADV 1;
FIG. 9 is a data diagram of the preservation performance test of the antifreeze type virus sampling preservation solution of the invention, wherein the first line from top to bottom is ADV3, and the ADV1-10 sample is preserved at-20 ℃ for 7 days; the second thread is ADV 5; the third line is ADV 7; the fourth line is ADV 1; the fifth line is ADV 4; the sixth line is ADV 2; the seventh line is ADV 9; the eighth line is ADV 8; the ninth line is ADV 6; the tenth line is ADV 10;
FIG. 10 is a data diagram of the ADV1-10 sample at the preservation temperature of-30 ℃ for 7 days in the preservation performance test of the antifreeze virus sampling preservation solution provided by the invention, wherein the first line from top to bottom in the data diagram is ADV 2; the second thread is ADV 1; the third line is ADV 4; the fourth line is ADV 8; the fifth line is ADV 3; the sixth line is ADV 7; the seventh line is ADV 9; the eighth line is ADV 10; the ninth line is ADV 5; the tenth line is ADV 6;
FIG. 11 is a data diagram of the ADV1-10 sample at the preservation temperature of-40 ℃ for 7 days in the preservation performance test of the antifreeze virus sampling preservation solution provided by the invention, wherein the first line from top to bottom in the data diagram is ADV 9; the second thread is ADV 10; the third line is ADV 1; the fourth line is ADV 2; the fifth line is ADV 7; the sixth line is ADV 8; the seventh line is ADV 6; the eighth line is ADV 3; the ninth line is ADV 4; the tenth line is ADV 5;
FIG. 12 is a data diagram of the preservation temperature of the sample ADV1-10 at-80 ℃ for 7 days in the preservation performance test of the antifreeze virus sampling preservation solution provided by the invention, wherein the first line from top to bottom in the data diagram is ADV 9; the second thread is ADV 1; the third line is ADV 2; the fourth line is ADV 10; the fifth line is ADV 3; the sixth line is ADV 8; the seventh line is ADV 6; the eighth line is ADV 4; the ninth line is ADV 5; the tenth line is ADV 7;
FIG. 13 is a data diagram of ADV1-10 samples stored at-20 deg.C for 14 days in the antifreeze virus sampling preservation solution storage performance test, the first line from top to bottom in the data diagram is ADV 3; the second thread is ADV 6; the third line is ADV 8; the fourth line is ADV 5; the fifth line is ADV 10; the sixth line is ADV 7; the seventh line is ADV 9; the eighth line is ADV 4; the ninth line is ADV 1; the tenth line is ADV 2;
FIG. 14 is a data diagram of ADV1-10 samples stored at-30 ℃ for 14 days in the antifreeze virus sampling and storing liquid storage performance test, wherein the first line from top to bottom is ADV 2; the second thread is ADV 7; the third line is ADV 1; the fourth line is ADV 4; the fifth line is ADV 8; the sixth line is ADV 9; the seventh line is ADV 10; the eighth line is ADV 3; the ninth line is ADV 6; the tenth line is ADV 5;
FIG. 15 is a data diagram of ADV1-10 samples stored at-40 deg.C for 14 days in the antifreeze virus sampling preservation solution storage performance test, the first line from top to bottom in the data diagram is ADV 3; the second thread is ADV 4; the third line is ADV 8; the fourth line is ADV 1; the fifth line is ADV 7; the sixth line is ADV 5; the seventh line is ADV 6; the eighth line is ADV 2; the ninth line is ADV 10; the tenth line is ADV 9;
FIG. 16 is a data diagram of ADV1-10 samples stored at-80 ℃ for 14 days in the antifreeze virus sampling preservation solution storage performance test, wherein the first line from top to bottom is ADV 1; the second thread is ADV 2; the third line is ADV 9; the fourth line is ADV 4; the fifth line is ADV 3; the sixth line is ADV 10; the seventh line is ADV 8; the eighth line is ADV 5; the ninth line is ADV 6; the tenth line is ADV 7;
FIG. 17 is a data diagram of ADV1-10 samples stored at-20 deg.C for 28 days in the antifreeze virus sampling preservation solution storage performance test, the first line from top to bottom in the data diagram is ADV 9; the second thread is ADV 8; the third line is ADV 10; the fourth line is ADV 7; the fifth line is ADV 1; the sixth line is ADV 2; the seventh line is ADV 3; the eighth line is ADV 4; the ninth line is ADV 5; the tenth line is ADV 6;
FIG. 18 is a data diagram of ADV1-10 samples stored at-30 ℃ for 28 days in the antifreeze virus sampling and storing liquid storage performance test, wherein the first line from top to bottom is ADV 3; the second thread is ADV 2; the third line is ADV 6; the fourth line is ADV 5; the fifth line is ADV 4; the sixth line is ADV 1; the seventh line is ADV 8; the eighth line is ADV 10; the ninth line is ADV 9; the tenth line is ADV 7;
FIG. 19 is a data diagram of ADV1-10 samples stored at-40 deg.C for 28 days in the antifreeze virus sampling preservation solution storage performance test, the first line from top to bottom in the data diagram is ADV 9; the second thread is ADV 2; the third line is ADV 1; the fourth line is ADV 4; the fifth line is ADV 6; the sixth line is ADV 8; the seventh line is ADV 7; the eighth line is ADV 3; the ninth line is ADV 10; the tenth line is ADV 5;
FIG. 20 is a data diagram of ADV1-10 samples stored at-80 deg.C for 28 days in the antifreeze virus sampling preservation solution storage performance test, the first line from top to bottom in the data diagram is ADV 9; the second thread is ADV 10; the third line is ADV 8; the fourth line is ADV 7; the fifth line is ADV 1; the sixth line is ADV 2; the seventh line is ADV 4; the eighth line is ADV 6; the ninth line is ADV 3; the tenth line is ADV 5;
FIG. 21 is a data diagram of the ADV1-10 sample at the preservation temperature of-80 ℃ for 56 days in the preservation performance test of the antifreeze virus sampling preservation solution provided by the invention, wherein the first line from top to bottom in the data diagram is ADV 3; the second thread is ADV 6; the third line is ADV 9; the fourth line is ADV 10; the fifth line is ADV 4; the sixth line is ADV 5; the seventh line is ADV 8; the eighth line is ADV 2; the ninth line is ADV 7; the tenth line is ADV 1;
FIG. 22 is a data chart of 0 day at a preservation temperature of 2 ℃ to 8 ℃ of an EV1-10 sample in a preservation performance test of the antifreeze virus sampling preservation solution provided by the invention, wherein the first line from top to bottom in the data chart is EV 6; the second line is EV 8; the third wire is EV 3; the fourth line is EV 1; the fifth line is EV 4; the sixth line is EV 5; the seventh line is EV 7; the eighth line is EV 2; the ninth line is EV 10; the tenth line is EV 9;
FIG. 23 is a data chart of 0 day at-10 ℃ storage temperature of an EV1-10 sample in the antifreeze virus sampling storage fluid storage performance test provided by the invention, wherein the first line from top to bottom in the data chart is EV 7; the second line is EV 8; the third wire is EV 10; the fourth line is EV 9; the fifth line is EV 6; the sixth line is EV 3; the seventh line is EV 5; the eighth line is EV 4; the ninth line is EV 2; the tenth line is EV 1;
FIG. 24 is a data chart of 0 day at-20 ℃ storage temperature of an EV1-10 sample in the antifreeze virus sampling storage fluid storage performance test provided by the invention, wherein the first line from top to bottom in the data chart is EV 9; the second line is EV 8; the third wire is EV 10; the fourth line is EV 7; the fifth line is EV 5; the sixth line is EV 1; the seventh line is EV 3; the eighth line is EV 2; the ninth line is EV 4; the tenth line is EV 6;
FIG. 25 is a data chart of 0 day at-30 ℃ storage temperature of an EV1-10 sample in the antifreeze virus sampling storage fluid storage performance test provided by the invention, wherein the first line from top to bottom in the data chart is EV 9; the second line is EV 7; the third wire is EV 4; the fourth line is EV 3; the fifth line is EV 10; the sixth line is EV 8; the seventh line is EV 1; the eighth line is EV 2; the ninth line is EV 5; the tenth line is EV 6;
FIG. 26 is a data chart of 0 day at-40 ℃ storage temperature of an EV1-10 sample in the antifreeze virus sampling storage fluid storage performance test provided by the invention, wherein the first line from top to bottom in the data chart is EV 8; the second line is EV 10; the third wire is EV 5; the fourth line is EV 3; the fifth line is EV 9; the sixth line is EV 4; the seventh line is EV 7; the eighth line is EV 2; the ninth line is EV 1; the tenth line is EV 6;
FIG. 27 is a data chart of 0 day at-80 ℃ storage temperature of an EV1-10 sample in a preservation performance test of an antifreeze virus sampling preservation solution provided by the invention, wherein the first line from top to bottom in the data chart is EV 9; the second line is EV 4; the third wire is EV 8; the fourth line is EV 7; the fifth line is EV 5; the sixth line is EV 2; the seventh line is EV 3; the eighth line is EV 1; the ninth line is EV 6; the tenth line is EV 10;
FIG. 28 is a data graph of EV1-10 samples stored at 2 deg.C-8 deg.C for 7 days in the antifreeze virus sampling storage fluid storage performance test, the first line from top to bottom in the graph is EV 8; the second line is EV 3; the third wire is EV 7; the fourth line is EV 4; the fifth line is EV 2; the sixth line is EV 1; the seventh line is EV 5; the eighth line is EV 6; the ninth line is EV 9; the tenth line is EV 10;
FIG. 29 is a data chart of an EV1-10 sample at a preservation temperature of-10 ℃ for 7 days in a preservation performance test of the antifreeze virus sampling preservation solution provided by the invention, wherein the first line from top to bottom in the data chart is EV 10; the second line is EV 8; the third wire is EV 9; the fourth line is EV 4; the fifth line is EV 7; the sixth line is EV 3; the seventh line is EV 1; the eighth line is EV 2; the ninth line is EV 6; the tenth line is EV 5;
FIG. 30 is a data chart of EV1-10 samples stored at-20 deg.C for 7 days in the antifreeze virus sampling and storing liquid storage performance test provided by the present invention, in which the first line from top to bottom is EV 8; the second line is EV 9; the third wire is EV 1; the fourth line is EV 10; the fifth line is EV 4; the sixth line is EV 7; the seventh line is EV 3; the eighth line is EV 2; the ninth line is EV 6; the tenth line is EV 5;
FIG. 31 is a data chart of the EV1-10 sample at the preservation temperature of-30 ℃ for 7 days in the preservation performance test of the antifreeze virus sampling preservation solution provided by the invention, wherein the first line from top to bottom in the data chart is EV 9; the second line is EV 10; the third wire is EV 8; the fourth line is EV 3; the fifth line is EV 7; the sixth line is EV 2; the seventh line is EV 1; the eighth line is EV 4; the ninth line is EV 6; the tenth line is EV 5;
FIG. 32 is a data chart of EV1-10 samples stored at-40 deg.C for 7 days in the antifreeze virus sampling and storing liquid storage performance test, the first line from top to bottom in the chart is EV 9; the second line is EV 10; the third wire is EV 7; the fourth line is EV 8; the fifth line is EV 3; the sixth line is EV 4; the seventh line is EV 1; the eighth line is EV 2; the ninth line is EV 6; the tenth line is EV 5;
FIG. 33 is a data chart of the EV1-10 sample at the preservation temperature of-80 ℃ for 7 days in the preservation performance test of the antifreeze virus sampling preservation solution provided by the invention, wherein the first line from top to bottom in the data chart is EV 4; the second line is EV 1; the third wire is EV 2; the fourth line is EV 3; the fifth line is EV 9; the sixth line is EV 10; the seventh line is EV 8; the eighth line is EV 7; the ninth line is EV 6; the tenth line is EV 5;
FIG. 34 is a data chart of EV1-10 samples stored at-20 deg.C for 14 days in the antifreeze virus sampling and storing liquid storage performance test, the first line from top to bottom in the data chart being EV 4; the second line is EV 8; the third wire is EV 3; the fourth line is EV 10; the fifth line is EV 9; the sixth line is EV 2; the seventh line is EV 7; the eighth line is EV 6; the ninth line is EV 5; the tenth line is EV 1;
FIG. 35 is a data chart of EV1-10 samples stored at-30 deg.C for 14 days in the antifreeze virus sampling and storing liquid storage performance test, the first line from top to bottom in the chart being EV 3; the second line is EV 4; the third wire is EV 5; the fourth line is EV 9; the fifth line is EV 7; the sixth line is EV 10; the seventh line is EV 8; the eighth line is EV 6; the ninth line is EV 2; the tenth line is EV 1;
FIG. 36 is a data chart of EV1-10 samples stored at-40 deg.C for 14 days in the antifreeze virus sampling and storing liquid storage performance test, the first line from top to bottom in the data chart being EV 8; the second line is EV 2; the third wire is EV 7; the fourth line is EV 1; the fifth line is EV 9; the sixth line is EV 10; the seventh line is EV 5; the eighth line is EV 3; the ninth line is EV 4; the tenth line is EV 6;
FIG. 37 is a data chart of EV1-10 samples stored at-80 deg.C for 14 days in the antifreeze virus sampling and storing liquid storage performance test, the first line from top to bottom in the data chart is EV 8; the second line is EV 1; the third wire is EV 4; the fourth line is EV 2; the fifth line is EV 3; the sixth line is EV 9; the seventh line is EV 10; the eighth line is EV 7; the ninth line is EV 6; the tenth line is EV 5;
FIG. 38 is a data graph of a sample EV1-10 stored at a temperature of-20 ℃ for 28 days in a preservation performance test of the antifreeze virus sampling preservation solution provided by the present invention, wherein the first line from top to bottom in the graph is EV 9; the second line is EV 7; the third wire is EV 2; the fourth line is EV 10; the fifth line is EV 6; the sixth line is EV 1; the seventh line is EV 5; the eighth line is EV 8; the ninth line is EV 4; the tenth line is EV 3;
FIG. 39 is a data chart of the EV1-10 sample at the preservation temperature of-30 ℃ for 28 days in the preservation performance test of the antifreeze virus sampling preservation solution provided by the invention, wherein the first line from top to bottom in the data chart is EV 9; the second line is EV 1; the third wire is EV 3; the fourth line is EV 7; the fifth line is EV 10; the sixth line is EV 5; the seventh line is EV 6; the eighth line is EV 2; the ninth line is EV 8; the tenth line is EV 4;
FIG. 40 is a data chart of the EV1-10 sample at the preservation temperature of-40 ℃ for 28 days in the preservation performance test of the antifreeze virus sampling preservation solution provided by the invention, wherein the first line from top to bottom in the data chart is EV 3; the second line is EV 4; the third wire is EV 10; the fourth line is EV 8; the fifth line is EV 7; the sixth line is EV 9; the seventh line is EV 1; the eighth line is EV 2; the ninth line is EV 6; the tenth line is EV 5;
FIG. 41 is a data chart of EV1-10 samples stored at-80 deg.C for 28 days in the antifreeze virus sampling and storing liquid storage performance test, the first line from top to bottom in the chart is EV 5; the second line is EV 8; the third wire is EV 6; the fourth line is EV 3; the fifth line is EV 1; the sixth line is EV 7; the seventh line is EV 10; the eighth line is EV 4; the ninth line is EV 9; the tenth line is EV 2;
FIG. 42 is a data chart of an EV1-10 sample at a preservation temperature of-80 ℃ for 56 days in a preservation performance test of the antifreeze virus sampling preservation solution provided by the invention, wherein the first line from top to bottom in the data chart is EV 9; the second line is EV 10; the third wire is EV 1; the fourth line is EV 2; the fifth line is EV 3; the sixth line is EV 8; the seventh line is EV 4; the eighth line is EV 7; the ninth line is EV 6; the tenth line is EV 5.
Detailed Description
The technical solutions in the embodiments of the present invention are clearly and completely described below, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all embodiments.
The rapid and accurate etiology diagnosis is one of the important means for controlling the spread of epidemic situation. By adopting a real-time fluorescent quantitative PCR (RT-PCR) method, the novel coronavirus nucleic acid can be detected in nasopharyngeal swab, sputum and other specimens of lower respiratory tract secretion, blood, excrement, urine and the like of an infected person. 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. In order to solve the technical problems, the invention provides an anti-freezing virus sampling tube which comprises a swab and a collecting tube and is characterized by further comprising a preservation solution.
In the embodiment of the invention, the anti-freezing virus sampling tube comprises a swab and a collecting tube, and further comprises a preservation solution.
The anti-freezing type virus sampling and preserving fluid mainly comprises an acid-base indicator, pH buffer salt, a protein denaturant, an anti-freezing agent, antibiotics and a solvent;
the acid-base indicator comprises phenol red; the pH buffer salt comprises disodium hydrogen phosphate dodecahydrate and potassium dihydrogen phosphate; protein denaturants including guanidinium isothiocyanate and ethylphenylpolyethylene glycol; anti-freezing agents include glycerol and ethylene glycol; antibiotics include gentamicin; the solvent comprises purified water.
Every 30L of the preservation solution contains 0.001-0.004% of phenol red, 2.0-3.0 mm of disodium hydrogen phosphate dodecahydrate, 0.2-1.0 mm of potassium dihydrogen phosphate, 0.4-4 m/L of guanidine isothiocyanate, 0.3-1% of ethyl phenyl polyethylene glycol, 10-40% of glycerol, 10-40% of ethylene glycol, 0.3-3g of gentamicin and purified water with the constant volume of 30L.
The pH value of the preservation solution is 7.0-8.0.
The preparation method of the preservation solution comprises the following steps: mixing disodium hydrogen phosphate dodecahydrate, potassium dihydrogen phosphate, guanidine isothiocyanate and purified water uniformly, adding ethylphenyl polyethylene glycol, glycerol and ethylene glycol, mixing uniformly, adding phenol red, and mixing uniformly to obtain the preservation solution.
The technical effects of the antifreeze virus sampling and preserving fluid and the sampling tube of the present invention will be further described with reference to the following specific examples, but the specific implementation methods mentioned in these examples are only illustrative and explanatory of the technical solution of the present invention, and do not limit the implementation scope of the present invention, and all modifications and substitutions based on the above principles should be within the protection scope of the present invention.
Example 1
Weighing 30L of preservation solution according to the proportion that each 30L of preservation solution contains 0.001% of phenol red, 2.0mm of disodium hydrogen phosphate dodecahydrate, 0.2mm of monopotassium phosphate, 0.4m/L of guanidine isothiocyanate, 0.3% of ethyl phenyl polyethylene glycol, 30% of glycerol, 30% of ethylene glycol, 0.3g of gentamicin and purified water with constant volume of 30L; mixing disodium hydrogen phosphate dodecahydrate, potassium dihydrogen phosphate, guanidine isothiocyanate and purified water uniformly, adding ethylphenyl polyethylene glycol, glycerol and ethylene glycol, mixing uniformly, adding phenol red, and mixing uniformly to obtain the preservation solution.
Example 2
Weighing 30L of preservation solution according to the proportion that each 30L of preservation solution contains 0.003 percent of phenol red, 2.4mm of disodium hydrogen phosphate dodecahydrate, 0.4mm of monopotassium phosphate, 0.8m/L of guanidine isothiocyanate, 0.5 percent of ethyl phenyl polyethylene glycol, 40 percent of glycerin, 40 percent of ethylene glycol, 0.6g of gentamicin and purified water with constant volume of 30L; mixing disodium hydrogen phosphate dodecahydrate, potassium dihydrogen phosphate, guanidine isothiocyanate and purified water uniformly, adding ethylphenyl polyethylene glycol, glycerol and ethylene glycol, mixing uniformly, adding phenol red, and mixing uniformly to obtain the preservation solution.
Example 3
Weighing 30L of preservation solution according to the proportion that each 30L of preservation solution contains 0.004 percent of phenol red, 2.8mm of disodium hydrogen phosphate dodecahydrate, 0.8mm of monopotassium phosphate, 1.5m/L of guanidine isothiocyanate, 0.5 percent of ethyl phenyl polyethylene glycol, 40 percent of glycerin, 40 percent of ethylene glycol, 0.6g of gentamicin and purified water with constant volume of 30L; mixing disodium hydrogen phosphate dodecahydrate, potassium dihydrogen phosphate, guanidine isothiocyanate and purified water uniformly, adding ethylphenyl polyethylene glycol, glycerol and ethylene glycol, mixing uniformly, adding phenol red, and mixing uniformly to obtain the preservation solution.
Example 4
Weighing 30L of preservation solution according to the proportion that each 30L of preservation solution contains 0.004 percent of phenol red, 3.0mm of disodium hydrogen phosphate dodecahydrate, 1.0mm of monopotassium phosphate, 3m/L of guanidine isothiocyanate, 0.5 percent of ethyl phenyl polyethylene glycol, 40 percent of glycerin, 40 percent of glycol, 1.2g of gentamicin and purified water with the constant volume of 30L; mixing disodium hydrogen phosphate dodecahydrate, potassium dihydrogen phosphate, guanidine isothiocyanate and purified water uniformly, adding ethylphenyl polyethylene glycol, glycerol and ethylene glycol, mixing uniformly, adding phenol red, and mixing uniformly to obtain the preservation solution.
Further, the invention also makes systematic research on the antifreeze virus sampling and storing liquid and the process conditions in the sampling pipe, and only the following test scheme for obviously influencing the effect of the antifreeze virus sampling and storing liquid by changing the process conditions is explained, and the process conditions of the embodiment 3 are taken as the basis, and the concrete results are shown in comparative examples 1-2:
comparative example 1
Weighing 30L of preservation solution according to the proportion that each 30L of preservation solution contains 0.004 percent of phenol red, 2.8mm of disodium hydrogen phosphate dodecahydrate, 0.8mm of monopotassium phosphate, 1.5m/L of guanidine isothiocyanate, 0.5 percent of ethyl phenyl polyethylene glycol, 40 percent of glycerin, 0.6g of gentamicin and purified water with constant volume of 30L; mixing disodium hydrogen phosphate dodecahydrate, potassium dihydrogen phosphate, guanidine isothiocyanate and purified water uniformly, adding ethylphenyl polyethylene glycol and glycerol, mixing uniformly, adding phenol red, and mixing uniformly to obtain the preservation solution.
Comparative example 2
Weighing 30L of preservation solution according to the proportion that each 30L of preservation solution contains 0.004 percent of phenol red, 2.8mm of disodium hydrogen phosphate dodecahydrate, 0.8mm of monopotassium phosphate, 1.5m/L of guanidine isothiocyanate, 0.5 percent of ethyl phenyl polyethylene glycol, 40 percent of glycol, 0.6g of gentamicin and purified water with the constant volume of 30L; mixing disodium hydrogen phosphate dodecahydrate, potassium dihydrogen phosphate, guanidine isothiocyanate and purified water uniformly, adding ethylphenyl polyethylene glycol and ethylene glycol, mixing uniformly, adding phenol red, and mixing uniformly to obtain the preservation solution.
The preservation effect of the preservation solution prepared in the examples 1 to 4 and the comparative examples 1 to 2 of the invention on the virus samples is detected, and a control group is set, wherein the control group comprises: the method adopts a patent CN 111690640A, wherein the concentration of guanidine hydrochloride is 5mol/L, the concentration of tris (hydroxymethyl) aminomethane hydrochloride is 60mmol/L, the concentration of ethylene diamine tetraacetic acid is 3mmol/L, and the volume percentage of isopropanol is 20%.
Taking an inactivated adenovirus sample (ADV, DNA virus) as a model, respectively adding the ADV virus sample into the virus preservation solution, then diluting the sample by 10 times, 100 times and 1000 times by using normal saline, and then performing a PCR nucleic acid sensitivity experiment on the sample, and determining the CT value, wherein the determination result is shown in Table 1:
TABLE 1
The preservation solutions of examples 1 to 4 of the present invention, comparative examples 1 to 2, and control were stored at different temperatures for 12 hours and observed whether the appearance state was frozen, and the observation results are shown in table 2:
TABLE 2
The preservation performance of the preservation solution prepared in example 1 of the present invention was tested.
Detecting a sample: an inactivated adenovirus sample (ADV, obtained from the viral disease of the Chinese disease control center), and an inactivated enterovirus sample (EV, obtained from the viral disease of the Chinese disease control center); reagent: a frozen virus sampling tube (batch number: 20080595), a magnetic bead method virus DNA/RNA rapid extraction kit (batch number: 20061083-08F1), an enterovirus general type RNA detection kit (a fluorescent PCR method) (batch number: 200706023), and an adenovirus nucleic acid detection kit (a fluorescent PCR method) (batch number: 200710047) of Baishi medical science and technology Limited in Jiangsu province; the instrument comprises the following steps: BZK-96 model automatic nucleic acid extractor, ABI7500 fluorescence PCR instrument; the verification method comprises the following steps: 20 (3 ml/tube) anti-freezing type virus sampling tubes (batch number: 20080595) are mixed with the anti-freezing type virus sampling tubes by taking an inactivated adenovirus sample (from being given by a virus disease in the Chinese disease control center) and an inactivated enterovirus sample (from being given by a virus disease in the Chinese disease control center) as samples, wherein each virus is respectively added into 10 tubes, and each tube is added with 100 mu l of the sample. The adenovirus samples were numbered as follows: ADV1, ADV2, ADV3 … … ADV10, and the numbers of the enterotoxin samples after loading are: EV1, EV2, EV3 … … EV10 were taken out at different time points at a predetermined storage temperature and tested on ABI7500 according to the instructions of the corresponding virus test kit. The time point settings are as in table 3:
TABLE 3
The amplification results for the inactivated adenovirus samples are shown in tables 4-8 below:
TABLE 4 preservation of inactivated adenovirus sample amplification test data at different temperatures (day 0)
| ADV1 | ADV2 | ADV3 | ADV4 | ADV5 | ADV6 | ADV7 | ADV8 | ADV9 | ADV10 | AVERAGE | |
| 2-8℃ | 22.27 | 22.21 | 22.66 | 22.67 | 23.21 | 23.08 | 22.57 | 22.44 | 22.64 | 22.61 | 22.64 |
| -10℃ | 22.45 | 22.51 | 22.52 | 22.42 | 23.12 | 23.20 | 21.86 | 21.92 | 22.64 | 22.63 | 22.53 |
| -20℃ | 21.99 | 21.97 | 22.18 | 21.98 | 22.47 | 22.67 | 22.58 | 22.54 | 22.15 | 22.35 | 22.29 |
| -30℃ | 22.66 | 22.71 | 22.69 | 22.63 | 22.84 | 22.82 | 22.39 | 22.32 | 22.78 | 22.74 | 22.66 |
| -40℃ | 22.26 | 22.41 | 22.87 | 22.90 | 23.42 | 22.42 | 23.21 | 22.78 | 22.66 | 22.25 | 22.72 |
| -80℃ | 22.63 | 22.81 | 22.24 | 22.12 | 22.64 | 22.66 | 22.46 | 22.46 | 22.73 | 22.76 | 22.55 |
TABLE 5 preservation of inactivated adenovirus sample amplification test data at different temperatures (7 days)
| ADV1 | ADV2 | ADV3 | ADV4 | ADV5 | ADV6 | ADV7 | ADV8 | ADV9 | ADV10 | AVERAGE | |
| 2-8℃ | 22.41 | 22.46 | 21.53 | 21.69 | 22.23 | 22.22 | 22.28 | 22.26 | 22.62 | 22.73 | 22.24 |
| -10℃ | 23.62 | 23.59 | 23.40 | 23.13 | 22.61 | 22.69 | 22.49 | 22.65 | 22.60 | 22.60 | 22.94 |
| -20℃ | 22.54 | 22.59 | 22.07 | 22.55 | 22.39 | 22.82 | 22.41 | 22.77 | 22.73 | 22.85 | 22.57 |
| -30℃ | 22.14 | 22.12 | 22.40 | 22.23 | 22.93 | 23.05 | 22.43 | 22.36 | 22.51 | 22.53 | 22.47 |
| -40℃ | 22.13 | 22.16 | 22.43 | 22.54 | 22.59 | 22.42 | 22.16 | 22.23 | 21.96 | 22.04 | 22.27 |
| -80℃ | 21.96 | 22.04 | 22.18 | 22.38 | 22.43 | 22.34 | 22.58 | 22.19 | 21.83 | 22.17 | 22.21 |
TABLE 6 preservation of inactivated adenovirus sample amplification test data at different temperatures (14 days)
TABLE 7 preservation of inactivated adenovirus sample amplification test data at different temperatures (28 days)
| ADV1 | ADV2 | ADV3 | ADV4 | ADV5 | ADV6 | ADV7 | ADV8 | ADV9 | ADV10 | AVERAGE | |
| 2-8℃ | / | / | / | / | / | / | / | / | / | / | / |
| -10℃ | / | / | / | / | / | / | / | / | / | / | / |
| -20℃ | 22.14 | 22.23 | 22.40 | 22.46 | 23.27 | 23.40 | 21.91 | 21.82 | 21.71 | 21.87 | 22.32 |
| -30℃ | 22.50 | 21.87 | 21.86 | 22.05 | 21.98 | 21.91 | 22.65 | 22.50 | 22.60 | 22.55 | 22.25 |
| -40℃ | 21.93 | 21.90 | 22.05 | 21.94 | 22.21 | 21.95 | 22.02 | 22.00 | 21.71 | 22.13 | 21.98 |
| -80℃ | 21.93 | 22.21 | 22.40 | 22.21 | 23.06 | 23.27 | 21.82 | 21.81 | 21.80 | 21.80 | 22.23 |
TABLE 8 preservation of inactivated adenovirus sample amplification test data at different temperatures (56 days)
| ADV1 | ADV2 | ADV3 | ADV4 | ADV5 | ADV6 | ADV7 | ADV8 | ADV9 | ADV10 | AVERAGE | |
| 2-8℃ | / | / | / | / | / | / | / | / | / | / | / |
| -10℃ | / | / | / | / | / | / | / | / | / | / | / |
| -20℃ | / | / | / | / | / | / | / | / | / | / | / |
| -30℃ | / | / | / | / | / | / | / | / | / | / | / |
| -40℃ | / | / | / | / | / | / | / | / | / | / | / |
| -80℃ | 22.75 | 22.61 | 21.79 | 21.90 | 21.92 | 21.83 | 22.71 | 22.11 | 21.83 | 21.88 | 22.13 |
The amplification results of the inactivated enterovirus samples are shown in table 9:
TABLE 9 preservation of inactivated enterovirus sample amplification test data at different temperatures (day 0)
TABLE 10 preservation of inactivated enterovirus sample amplification test data at different temperatures (7 days)
| EV1 | EV2 | EV3 | EV4 | EV5 | EV6 | EV7 | EV8 | EV9 | EV10 | AVERAGE | |
| 2-8℃ | 22.35 | 22.33 | 22.12 | 22.32 | 22.35 | 22.37 | 22.31 | 22.02 | 22.47 | 22.58 | 22.32 |
| -10℃ | 22.84 | 22.91 | 22.83 | 22.74 | 22.95 | 22.93 | 22.76 | 22.58 | 22.60 | 22.54 | 22.77 |
| -20℃ | 22.42 | 22.66 | 22.52 | 22.47 | 23.24 | 23.21 | 22.49 | 22.29 | 22.36 | 22.45 | 22.61 |
| -30℃ | 22.59 | 22.58 | 22.45 | 22.62 | 22.68 | 22.62 | 22.49 | 22.38 | 22.20 | 22.25 | 22.49 |
| -40℃ | 22.76 | 22.80 | 22.72 | 22.73 | 23.05 | 22.88 | 22.57 | 22,67 | 22.39 | 22.44 | 22.70 |
| -80℃ | 22.48 | 22.49 | 22.53 | 22.44 | 23.35 | 22.74 | 22.70 | 22.65 | 22.57 | 22.63 | 22.66 |
TABLE 11 preservation of inactivated enterovirus sample amplification test data at different temperatures (14 days)
| EV1 | EV2 | EV3 | EV4 | EV5 | EV6 | EV7 | EV8 | EV9 | EV10 | AVERAGE | |
| 2-8℃ | / | / | / | / | / | / | / | / | / | / | / |
| -10℃ | / | / | / | / | / | / | / | / | / | / | / |
| -20℃ | 22.61 | 22.33 | 22.16 | 21.98 | 22.50 | 22.48 | 22.33 | 22.10 | 22.25 | 22.17 | 22.29 |
| -30℃ | 22.62 | 22.50 | 21.90 | 22.16 | 22.17 | 22.41 | 22.21 | 22.36 | 22.17 | 22.29 | 22.28 |
| -40℃ | 22.41 | 22.32 | 22.62 | 22.68 | 22.61 | 22.70 | 22.36 | 22.23 | 22.44 | 22.49 | 22.49 |
| -80℃ | 22.55 | 22.58 | 22.63 | 22.56 | 23.37 | 23.30 | 22.69 | 22.48 | 22.65 | 22.66 | 22.75 |
TABLE 12 preservation of inactivated enterovirus sample amplification test data at different temperatures (28 days)
| EV1 | EV2 | EV3 | EV4 | EV5 | EV6 | EV7 | EV8 | EV9 | EV10 | AVERAGE | |
| 2-8℃ | / | / | / | / | / | / | / | / | / | / | / |
| -10℃ | / | / | / | / | / | / | / | / | / | / | / |
| -20℃ | 22.65 | 22.60 | 22.75 | 22.69 | 22.66 | 22.64 | 22.53 | 22.67 | 22.52 | 22.60 | 22.63 |
| -30℃ | 22.49 | 22.64 | 22.49 | 22.83 | 22.58 | 22.61 | 22.53 | 22.67 | 22.38 | 22.54 | 22.58 |
| -40℃ | 22.45 | 22.60 | 22.12 | 22.13 | 22.71 | 22.67 | 22.28 | 22.26 | 22.28 | 22.16 | 22.37 |
| -80℃ | 22.64 | 22.78 | 22.58 | 22.72 | 22.26 | 22.29 | 22.67 | 22.26 | 22.72 | 22.67 | 22.56 |
TABLE 13 preservation of inactivated enterovirus sample amplification test data at different temperatures (56 days)
| EV1 | EV2 | EV3 | EV4 | EV5 | EV6 | EV7 | EV8 | EV9 | EV10 | AVERAGE | |
| 2-8℃ | / | / | / | / | / | / | / | / | / | / | / |
| -10℃ | / | / | / | / | / | / | / | / | / | / | / |
| -20℃ | / | / | / | / | / | / | / | / | / | / | / |
| -30℃ | / | / | / | / | / | / | / | / | / | / | / |
| -40℃ | / | / | / | / | / | / | / | / | / | / | / |
| -80℃ | 22.74 | 22.80 | 22.81 | 22.88 | 23.12 | 22.98 | 22.88 | 22.87 | 22.58 | 22.59 | 22.83 |
The appearance of the virus preservation solution at different temperatures is shown in table 14:
TABLE 14 appearance of virus preservation solution at different temperatures
In summary, the antifreeze virus sampling and storing solution and the sampling tube provided in this embodiment, wherein the storing solution is inactivated, the virus completely loses infectivity in the storing process, the biological safety is high, the antifreeze virus sampling and storing solution is suitable for storing various types of virus samples, such as influenza virus, enterovirus, HIV virus, coronavirus, etc., the storage effect is good, the DNA/RNA of the virus is not degraded in the storing process, and the virus release rate is high; the preservation solution is low temperature resistant, can preserve samples for a long time in an environment of 30 ℃ below zero, is not frozen and cracked, has stable virus nucleic acid, and is beneficial to the preservation and transportation of samples in extremely cold areas in winter; the use is simple and convenient, microbial protein can be cracked and inactivated, secondary infection risk is effectively prevented, and the safety of transportation and detection personnel is guaranteed; the applicable sample range is wide: such as oral swabs, pharyngeal swabs, nasal swabs, anal swabs, cervical swabs, saliva, sputum, alveolar lavage, and the like, and can also be used to collect environmental samples.
The above description is only for the preferred embodiment of the present invention, but the scope of the present invention is not limited thereto, and any person skilled in the art should be considered to be within the technical scope of the present invention, and the technical solutions and the inventive concepts thereof according to the present invention should be equivalent or changed within the scope of the present invention.
Claims (9)
1. The anti-freezing type virus sampling and preserving fluid is characterized by mainly comprising an acid-base indicator, pH buffer salt, a protein denaturant, an anti-freezing agent, antibiotics and a solvent;
the acid-base indicator comprises phenol red; the pH buffer salt comprises disodium hydrogen phosphate dodecahydrate and potassium dihydrogen phosphate; protein denaturants including guanidinium isothiocyanate and ethylphenylpolyethylene glycol; anti-freezing agents include glycerol and ethylene glycol; antibiotics include gentamicin; the solvent comprises purified water.
2. The antifreeze virus sampling and storing solution of claim 1, wherein each 30L of the solution contains 0.001-0.004% of phenol red, 2.0-3.0 mm of disodium hydrogen phosphate dodecahydrate, 0.2-1.0 mm of potassium dihydrogen phosphate, 0.4-4 m/L of guanidinium isothiocyanate, 0.3-1% of ethylphenylpolyethylene glycol, 10-40% of glycerol, 10-40% of ethylene glycol, 0.3-3g of gentamicin, and purified water up to 30L.
3. The antifreeze virus sampling and preserving fluid of claim 1, wherein the pH value of the preserving fluid is 7.0 to 8.0.
4. The antifreeze virus sampling and preserving fluid of claim 1, wherein each 30L of the preserving fluid comprises 0.001% of phenol red, 2.0mm of disodium hydrogen phosphate dodecahydrate, 0.2mm of monopotassium phosphate, 0.4m/L of guanidine isothiocyanate, 0.3% of ethylphenylpolyethylene glycol, 30% of glycerol, 30% of ethylene glycol, 0.3g of gentamicin and purified water up to 30L in volume.
5. The antifreeze virus sampling and preserving fluid of claim 1, wherein each 30L of the preserving fluid comprises 0.003% of phenol red, 2.4mm of disodium hydrogen phosphate dodecahydrate, 0.4mm of monopotassium phosphate, 0.8m/L of guanidinium isothiocyanate, 0.5% of ethylphenylpolyethylene glycol, 40% of glycerol, 40% of ethylene glycol, 0.6g of gentamicin, and purified water up to 30L in volume.
6. The antifreeze virus sampling and preserving fluid of claim 1, wherein each 30L of the preserving fluid comprises 0.004% of phenol red, 2.8mm of disodium hydrogen phosphate dodecahydrate, 0.8mm of potassium dihydrogen phosphate, 1.5m/L of guanidine isothiocyanate, 0.5% of ethylphenylpolyethylene glycol, 40% of glycerol, 40% of ethylene glycol, 0.6g of gentamicin and purified water up to 30L in volume.
7. The antifreeze virus sampling and preserving fluid of claim 1, wherein each 30L of the preserving fluid comprises 0.004% of phenol red, 3.0mm of disodium hydrogen phosphate dodecahydrate, 1.0mm of potassium dihydrogen phosphate, 3m/L of guanidine isothiocyanate, 0.5% of ethylphenylpolyethylene glycol, 40% of glycerol, 40% of ethylene glycol, 1.2g of gentamicin and purified water up to 30L.
8. The antifreeze virus sampling preservation solution according to any one of claims 1 to 7, wherein the preparation method of the preservation solution comprises the following steps: mixing disodium hydrogen phosphate dodecahydrate, potassium dihydrogen phosphate, guanidine isothiocyanate and purified water uniformly, adding ethylphenyl polyethylene glycol, glycerol and ethylene glycol, mixing uniformly, adding phenol red, and mixing uniformly to obtain the preservation solution.
9. A sampling tube comprising a swab and a collecting tube, wherein the sampling tube further comprises the antifreeze virus sampling and preserving fluid as set forth in any one of claims 1 to 8.
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