CN109371008B - Nucleic acid extraction amplification detection kit - Google Patents
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- CN109371008B CN109371008B CN201811247122.5A CN201811247122A CN109371008B CN 109371008 B CN109371008 B CN 109371008B CN 201811247122 A CN201811247122 A CN 201811247122A CN 109371008 B CN109371008 B CN 109371008B
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Abstract
The invention provides a nucleic acid extraction amplification detection kit, which consists of a lysate kit and a PCR tube. The nucleic acid lysis solution is contained in the lysis solution reagent box, after the sample to be detected is added with the nucleic acid lysis solution, the virus nucleic acid is released under the action of the nucleic acid lysis solution, the purification is realized, and after the sample is added with the nucleic acid lysis solution, the obtained substance can be directly added into a PCR tube to realize the PCR detection. The reagent of the frozen dry powder PCR amplification system directly used for fluorescent quantitative PCR measurement is placed in the PCR tube, and the reagent of the frozen dry powder PCR amplification system is an obtained product prepared by taking PCR reaction buffer solution, DNA polymerase, reaction specific primers, probes and freeze-drying protective agents as raw materials through a freeze-drying process.
Description
Technical Field
The invention relates to the technical field of biological detection, in particular to a nucleic acid extraction amplification detection kit and a detection method thereof.
Background
The real-time fluorescent quantitative PCR technology is widely applied to a plurality of fields such as genetic disease molecular diagnosis, clinical examination, animal and plant import and export quarantine, food safety monitoring, soil microorganism detection, paternity test and the like. Because samples such as blood, food, soil and the like contain a large amount of inhibiting factors such as hemoglobin, methemoglobin, lactoferrin, humic acid and the like, the Taq DNA polymerase has obvious inhibiting effect on the conventional Taq DNA polymerase. Therefore, nucleic acids must be isolated from these test samples and then used for PCR amplification. Nucleic acid extraction is the first step of nucleic acid detection, and is also one of the key methods in molecular biology. Provides a basis for downstream nucleic acid detection, and the quality and integrity of extraction directly influence clinical research or diagnosis.
The general nucleic acid extraction process comprises two steps: and (4) cracking and purifying the sample. The traditional extraction reagent has relatively low extraction rate and complex steps, brings certain influence on extraction work, and is difficult to obtain qualified nucleic acid samples. The magnetic bead method is a nucleic acid extraction method developed in recent years, and the extracted nucleic acid has high purity and large yield. Obviously, the traditional method has the disadvantages of multiple operation steps, high cost, large sample requirement, easy cross contamination and inconvenience for customers to quickly extract and purify nucleic acid samples.
In addition, in the current real-time fluorescence PCR technology with the widest application of nucleic acid detection, most reagents are not ready-to-use, so that the problems of complicated operation, high labor and cost and the like of PCR detection generally exist, complicated configuration work is required before use, and errors and even failures of tests are easily caused. Experimental errors also arise from different personnel operations; certain reagents are not stable to storage at ambient temperature; the rapid extraction of nucleic acid by one-step method cannot be realized; PCR detection reagents need to be manually configured on site, multiple steps need to be manually marked, detection instruments at all stages are independent, and full-automatic treatment of a large number of test samples is not facilitated.
Disclosure of Invention
In view of the above drawbacks of the prior art, the present invention provides a nucleic acid amplification detection kit and a detection method thereof. The nucleic acid extraction amplification detection kit and the detection method thereof adopt a real-time fluorescence PCR reaction system for directly extracting and amplifying virus nucleic acid in a reaction detection clinical sample, a sample to be detected is added into a lysis solution to realize nucleic acid lysis and purification, the product after lysis and purification can be directly subjected to PCR detection, and the amplification reaction and PCR detection are realized by using a ready-to-use freeze-dried powder reagent without preparation before detection, the bottleneck problem of PCR amplification depending on purified nucleic acid DNA as a template is thoroughly solved by detection, and the kit is suitable for various samples including blood, serum, plasma, saliva, urine, interstitial fluid, semen, secretion, pus, respiratory fluid, mucus and the like.
The invention provides a nucleic acid extraction amplification detection kit, which is characterized by comprising: the kit comprises a lysis solution kit and a PCR tube, wherein nucleic acid lysis solution is contained in the lysis solution kit, and virus nucleic acid is released under the action of the nucleic acid lysis solution; the reagent of the frozen dry powder PCR amplification system directly used for fluorescent quantitative PCR measurement is placed in the PCR tube, and the reagent of the frozen dry powder PCR amplification system is an obtained product prepared by taking PCR reaction buffer solution, DNA polymerase, reaction specific primers, probes and freeze-drying protective agents as raw materials through a freeze-drying process.
The nucleic acid lysate comprises GuHCl, NaOH, ethylene diamine tetraacetic acid, trihydroxymethyl aminomethane, NaCl, polyethylene glycol octyl phenyl ether, a surfactant, phenol and isoamylol. Wherein the concentration of GuHCl is 5.6-5.85g/10 ML; 0.03-0.04g of ethylenediamine tetraacetic acid/10 ML; 0.06-0.07g/10ML of trihydroxymethyl aminomethane; NaCl 0.01-0.015g/10 ML; 2-10% of polyethylene glycol octyl phenyl ether (volume ratio); 0.075-0.1% by volume of a surfactant; adding 1-2.5% (volume ratio) of phenol and isoamyl alcohol mixed solution into the nucleic acid cracking solution, wherein the volume ratio of phenol to isoamyl alcohol is 20:1-25: 1.
The surfactant adopts any one of the following components: sodium dodecyl sulfate, cetyl methyl triamine bromide, polyvinylpyrrolidone and sodium lauroyl ammonia acid.
The reagent for the frozen dry powder PCR amplification system is prepared by adopting the following method: mixing PCR reaction buffer solution, DNA polymerase, reaction specific primer, probe and freeze-drying protective agent solution according to a predetermined proportion, and freeze-drying.
Wherein the freeze drying is divided into three stages of pre-freezing, main freeze drying and reinforced drying, the first stage is pre-freezing, the temperature is slowly reduced to-35 to-40 ℃, the time length of temperature reduction is more than 1 hour, and the temperature is kept for 2.5 hours under the condition of the temperature; the second stage is a main freeze-drying stage, the temperature is maintained at-15 to-20 ℃ for 5 hours, wherein the air pressure is maintained at 0.1 mbar; the third stage is enhanced drying, the temperature is kept at 15-25 ℃, the drying time is 2 hours, and the air pressure is kept at 0.1 mbar.
Wherein, the freeze-drying protective agent adopted for preparing the reagent of the freeze-dried powder PCR amplification system comprises: mannitol, polyethylene glycol, maltose, ascorbic acid, and polyvinylpyrrolidone. Wherein, the addition amount of mannitol is 3-5g/10 ML; the adding amount of the polyethylene glycol is 2.5-3g/10 ML; the addition amount of maltose is 15-18g/10 ML; the addition amount of ascorbic acid is 0.4-0.6g/10 ML; polyvinylpyrrolidone 0.8-1.5 g/ML.
Wherein, deionized water added with pyrophosphatase is used for preparing the freeze-drying protective agent solution. Wherein the adding amount of the pyrosulfurylase is 1.1-1.6g/10 ML.
Wherein, the PCR reaction buffer solution adopted for preparing the reagent of the frozen dry powder PCR amplification system comprises: deoxyribonucleoside triphosphates, magnesium chloride.
Wherein, the reagent of the frozen dry powder PCR amplification system also contains transition metal sulfide.
The invention further claims a method for detecting nucleic acid by using the nucleic acid extraction amplification detection kit.
The invention enhances the effectiveness and purification degree of nucleic acid cracking, does not need nucleic acid precipitation, can directly detect the cracking solution, realizes amplification reaction and PCR detection by using a ready-to-use freeze-dried powder reagent, does not need to be prepared before detection, thoroughly solves the bottleneck problem of PCR amplification by using purified nucleic acid DNA as a template, and is suitable for various samples such as blood, serum, plasma, saliva, urine, interstitial fluid, semen, secretion, pus, respiratory fluid, mucus and the like.
Detailed Description
The technical scheme of the invention is further specifically described by the following embodiments.
The invention provides a nucleic acid extraction amplification detection kit, which consists of a lysate kit and a PCR tube. The nucleic acid lysis solution is contained in the lysis solution reagent box, after the sample to be detected is added with the nucleic acid lysis solution, the virus nucleic acid is released under the action of the nucleic acid lysis solution, the purification is realized, and after the sample is added with the nucleic acid lysis solution, the obtained substance can be directly added into a PCR tube to realize the PCR detection. The reagent of the frozen dry powder PCR amplification system directly used for fluorescent quantitative PCR measurement is placed in the PCR tube, and the reagent of the frozen dry powder PCR amplification system is an obtained product prepared by taking PCR reaction buffer solution, DNA polymerase, reaction specific primers, probes and freeze-drying protective agents as raw materials through a freeze-drying process.
(I) nucleic acid lysis solution
Specifically, the nucleic acid lysate comprises GuHCl, ethylene diamine tetraacetic acid, tris (hydroxymethyl) aminomethane, NaCl, polyethylene glycol octylphenyl ether, a surfactant, phenol and isoamyl alcohol. Wherein the concentration of GuHCl is 5.6-5.85g/10ML, preferably 5.75g/10 ML. Ethylenediaminetetraacetic acid 0.03-0.04g/10ML, preferably 0.038g/10 ML. Tris 0.06-0.07g/10ML, preferably 0.062g/10 ML. NaCl 0.01-0.015g/10ML, preferably 0.011g/10 ML. 2-10% of polyethylene glycol octyl phenyl ether (volume ratio), preferably 2.7%. The surfactant may be mixed with any one or more of the following: sodium dodecyl sulfate, cetyl methyl triamine bromide, polyvinylpyrrolidone and sodium lauroyl ammonia acid; 0.075-0.1% (by volume), preferably 0.85% of surfactant. Adding 1-2.5% (volume ratio) of phenol and isoamyl alcohol mixed solution into the nucleic acid cracking solution, wherein the volume ratio of phenol to isoamyl alcohol is 20:1-25: 1. The pH of the lysate was 4-4.5. In order to realize the subsequent direct PCR detection of the product obtained by nucleic acid cracking, the lysis solution can generate a nucleic acid product with high purity and high integrity, completely removes components such as protein, polysaccharide and the like in cells, has no residue of reagents and fluorescence inhibition effect, and retains the integrity of the base sequence of the nucleic acid molecule. In the cracking process, the GuHCl and NaCl mixed solution provides an environment for promoting the cracking release of nucleic acid, and the effect of protecting the nucleic acid from being degraded by nuclease is outstanding; the surfactant, protein and polysaccharide form insoluble precipitate to separate impurity from nucleic acid; phenol, isoamyl alcohol further facilitates the separation of the protein from the nucleic acid components, wherein phenol denatures the protein components, the denatured protein remains in the organic phase and the nucleic acids remain in the aqueous phase, and isoamyl alcohol facilitates the separation of the organic phase from the aqueous phase; the lysate does not contain nucleic acid precipitation components such as ethanol, isopropanol and the like, so the nucleic acid components are kept in the lysate and can be directly subjected to PCR detection. Wherein, GuHCl, NaCl, surfactant active, phenol, isoamyl alcohol concentration and lysate PH value have all optimized on the basis of the experiment, thus realize the maximization of nucleic acid extraction output.
Examples 1 to 3 of the lysate realized according to the present invention and the experimental effects thereof are specifically described below.
Example 1:
preparing 40ML of nucleic acid lysate according to the scheme of the invention, wherein the nucleic acid lysate comprises 5.6g/10ML of GuHCl, 0.03g/10ML of ethylene diamine tetraacetic acid, 0.06g/10ML of tris (hydroxymethyl) aminomethane, 0.01g/10ML of NaCl and 2% (volume ratio) of polyethylene glycol octyl phenyl ether, and the surfactant can adopt sodium dodecyl sulfate, 0.075% (volume ratio) of the surfactant and 1% (volume ratio) of a mixed solution of phenol and isoamyl alcohol, wherein the volume ratio of phenol to isoamyl alcohol is 20:1, and the pH value of the lysate is adjusted to 4.
Example 2
Preparing 40ML of nucleic acid lysate according to the scheme of the invention, wherein the nucleic acid lysate comprises 5.85g/10ML of GuHCl, 0.04g/10ML of ethylene diamine tetraacetic acid, 0.07g/10ML of tris (hydroxymethyl) aminomethane, 0.015g/10ML of NaCl and 10% (volume ratio) of polyethylene glycol octyl phenyl ether, the surfactant can be a mixture of polyvinylpyrrolidone and sodium lauroyl amide, the surfactant is 0.1% (volume ratio), 2.5% (volume ratio) of a mixed solution of phenol and isoamyl alcohol, wherein the volume ratio of phenol to isoamyl alcohol is 25:1, and the pH value of the lysate is adjusted to be 4.5.
Example 3
Preparing 40ML of nucleic acid lysate according to the scheme of the invention, wherein the nucleic acid lysate comprises 5.75g/10ML of GuHCl, 0.038g/10ML of ethylene diamine tetraacetic acid, 0.062g/10ML of tris (hydroxymethyl) aminomethane, 0.011g/10ML of NaCl, 2.7% (volume ratio) of polyethylene glycol octyl phenyl ether, the surfactant is cetyl methyl triamine bromide, the surfactant is 0.85% (volume ratio), 2.5% (volume ratio) of a mixed solution of phenol and isoamyl alcohol is added into the nucleic acid lysate, the volume ratio of phenol to isoamyl alcohol is 22:1, and the pH value of the lysate is adjusted to be 4.
And (3) taking 0.5ML of the nucleic acid lysate of the three embodiments respectively, taking 0.1ML of the virus plasma sample, mixing the sample and the lysate into a centrifugal tube, and inverting the centrifugal tube for 3-5 times to ensure that the nucleic acid is fully cracked and released. The yield of the nucleic acid extract in the lysate after the full reaction was measured using a NanoDrop 1000 microanalyte analyzer as follows: the results of 12.3. mu.g for example 1, 11.7. mu.g for example 2 and 14.8. mu.g for example 3 show that the yield of nucleic acid lysis is good, and the concentration of nucleic acid in the lysate can be directly measured by PCR.
(II) freezing dry powder PCR amplification system
A frozen dry powder PCR amplification system reagent directly used for fluorescent quantitative PCR measurement is placed in the PCR tube, a sample to be measured is added into a lysate and is fully cracked to obtain a liquid phase product, and RNA-removed water is directly added into the PCR tube to realize amplification and PCR fluorescence test. The reagent for the frozen dry powder PCR amplification system is prepared by adopting the following method: mixing PCR reaction buffer solution, DNA polymerase, reaction specific primer, probe and freeze-drying protective agent solution according to a predetermined proportion, and freeze-drying in three stages. Wherein, the first stage is pre-freezing, slowly cooling to-35 to-40 ℃, the cooling time is longer than 1 hour, and keeping for 2.5 hours under the temperature condition; the second stage is a freeze drying stage, the temperature is maintained at-15 to-20 ℃ for 5 hours, wherein the gas is in the freeze drying stageThe pressure was maintained at 0.1 mbar; the third stage is enhanced drying, the temperature is kept at 15-25 ℃, the drying time is 2 hours, and the air pressure is kept at 0.1 mbar. The freeze-drying protective agent is used for keeping the activity of enzyme required by PCR reaction in the freeze-drying process, and the freeze-drying protective agent used for preparing the frozen dry powder PCR amplification system reagent comprises: mannitol, polyethylene glycol, maltose, ascorbic acid, and polyvinylpyrrolidone; and preparing the components as the freeze-drying protective agent solution by using deionized water added with pyrophosphatase. Wherein, the addition amount of mannitol is 3-5g/10 ML; the adding amount of the polyethylene glycol is 2.5-3g/10 ML; maltose is added in an amount of 15-18g/10ML, preferably 16.4g/10 ML; the addition amount of ascorbic acid is 0.4-0.6g/10 ML; polyvinylpyrrolidone 0.8-1.5 g/ML. The adding amount of the pyrosulfatase is 1.1-1.6g/10 ML. The mannitol and the polyethylene glycol are mixed to be used as excipient, so that the excipient has good effect, the viscosity of the freeze-dried powder is kept, and the surface looseness is avoided. Maltose, ascorbic acid and polyvinylpyrrolidone are used as mixed type protective agents, and can replace lost water and protein to form hydrogen bonds in the freeze-drying process, so that the protective effect is achieved, and the activity of the protein is maintained. Preferably, the reagent for preparing the freeze-dried powder PCR amplification system further comprises transition metal sulfide MoS2The addition amount is 55 mug/10 ML, which can improve the sensitivity and specificity of detection. And the PCR reaction buffer solution adopted for preparing the reagent of the frozen dry powder PCR amplification system comprises: deoxyribonucleoside triphosphate and magnesium chloride, which are beneficial to the realization of amplification effect.
The reagents 1 to 3 of the frozen dry powder PCR amplification system realized according to the present invention and the experimental effects thereof are specifically described below.
Example 1
Preparing 10ML of a freeze-drying protective agent solution, wherein the specific components are as follows: the adding amount of mannitol is 3g/10ML, and the adding amount of polyethylene glycol is 2.5g/10 ML; maltose 15g/10ML, ascorbic acid 0.4g/10ML, polyvinylpyrrolidone 0.8g/ML and disulfatase 1.1g/10 ML. Mixing PCR reaction buffer solution, DNA polymerase, reaction specific primer, probe and the freeze-drying protective agent solution according to a preset proportion, and then carrying out freeze-drying in three stages. Wherein, the first stage is pre-freezing, slowly cooling to-35 to-40 ℃, the cooling time is longer than 1 hour, and keeping for 2.5 hours under the temperature condition; the second stage is a main freeze-drying stage, the temperature is maintained at-15 to-20 ℃ for 5 hours, wherein the air pressure is maintained at 0.1 mbar; the third stage is enhanced drying, the temperature is kept at 15-25 ℃, the drying time is 2 hours, and the air pressure is kept at 0.1 mbar.
Example 2
Preparing 10ML of a freeze-drying protective agent solution, wherein the specific components are as follows: the adding amount of mannitol is 5g/10ML, and the adding amount of polyethylene glycol is 3g/10 ML; maltose 18g/10ML, ascorbic acid 0.6g/10ML, polyvinylpyrrolidone 1.5g/ML and disulfatase 1.6g/10 ML. Mixing PCR reaction buffer solution mixed by deoxyribonucleoside triphosphate and magnesium chloride, DNA polymerase, reaction specific primer and probe with the freeze-drying protective agent solution according to a predetermined proportion, and freeze-drying in three stages. Wherein, the first stage is pre-freezing, slowly cooling to-35 to-40 ℃, the cooling time is longer than 1 hour, and keeping for 2.5 hours under the temperature condition; the second stage is a freeze drying stage, the temperature is maintained at-15 to-20 ℃, and the temperature is maintained for 5 hours, wherein the air pressure is maintained at 0.1 mbar; the third stage is enhanced drying, the temperature is kept at 15-25 ℃, the drying time is 2 hours, and the air pressure is kept at 0.1 mbar.
Example 3
Preparing 10ML of a freeze-drying protective agent solution, wherein the specific components are as follows: the addition amount of mannitol is 4.5g/10ML, and the addition amount of polyethylene glycol is 2.8g/10 ML; maltose 16.5g/10ML, ascorbic acid 0.55g/10ML, polyvinylpyrrolidone 1.1g/ML, and disulfatase 1.35g/10 ML. Mixing PCR reaction buffer solution, DNA polymerase, reaction specific primer, probe and the freeze-drying protective agent solution according to a preset proportion, and then carrying out freeze-drying in three stages. Wherein, the first stage is pre-freezing, slowly cooling to-35 to-40 ℃, the cooling time is longer than 1 hour, and keeping for 2.5 hours under the temperature condition; the second stage is a freeze drying stage, the temperature is maintained at-15 to-20 ℃, and the temperature is maintained for 5 hours, wherein the air pressure is maintained at 0.1 mbar; the third stage is enhanced drying, the temperature is kept at 15-25 ℃, the drying time is 2 hours, and the air pressure is kept at 0.1 mbar.
Dividing the reagents of the frozen dry powder PCR amplification system prepared in the three embodiments into 5 parts respectively, placing the reagents in an environment of 37 ℃ for 0, 3, 7, 10 and 20 days, then adding the test reagent prepared in the lysate embodiments 1-3, shaking and mixing the reagents uniformly, centrifuging the mixture, and then loading the sample on a machine for PCR test, wherein the test sensitivity of the samples of the embodiments 1-3 in 0, 3, 7 and 10 days is 104Sensitivity of 10 in 20 days5. It can be seen that the test sensitivity can be maintained for a longer time.
Therefore, the invention enhances the effectiveness and purification degree of nucleic acid cracking, does not need nucleic acid precipitation, can directly detect the cracking solution, realizes amplification reaction and PCR detection by using a ready-to-use freeze-dried powder reagent, does not need to be prepared before detection, thoroughly solves the bottleneck problem of PCR amplification by using purified nucleic acid DNA as a template, and is suitable for various samples such as blood, serum, plasma, saliva, urine, interstitial fluid, semen, secretion, pus, respiratory fluid, mucus and the like.
The above embodiments are only for illustrating the invention and are not to be construed as limiting the invention, and those skilled in the art can make various changes and modifications without departing from the spirit and scope of the invention, therefore, all equivalent technical solutions also belong to the scope of the invention, and the scope of the invention is defined by the claims.
Claims (1)
1. A nucleic acid extraction amplification detection kit is characterized by comprising: the kit comprises a lysate kit and a PCR tube, wherein nucleic acid lysate is contained in the lysate kit, viral nucleic acid is released and purified under the action of the nucleic acid lysate, and a product obtained after lysis and purification can be directly subjected to PCR detection; a frozen dry powder PCR amplification system reagent directly used for fluorescent quantitative PCR measurement is placed in the PCR tube, and the frozen dry powder PCR amplification system reagent is an obtained product prepared by taking a PCR reaction buffer solution, DNA polymerase, a reaction specific primer, a probe and a freeze-drying protective agent as raw materials through a freeze-drying process;
the nucleic acid lysate comprises GuHCl, NaOH, ethylene diamine tetraacetic acid, trihydroxymethyl aminomethane, NaCl, polyethylene glycol octyl phenyl ether, a surfactant, phenol and isoamylol; wherein the concentration of GuHCl is 5.75g/10 ML; ethylenediaminetetraacetic acid 0.038g/10 ML; 0.062g/10ML of tris (hydroxymethyl) aminomethane; NaCl 0.011g/10 ML; 2.7 percent of polyethylene glycol octyl phenyl ether (volume ratio); 0.85 percent of surfactant (volume ratio); adding 1-2.5% (volume ratio) of a mixed solution of phenol and isoamyl alcohol into the nucleic acid lysate, wherein the volume ratio of the phenol to the isoamyl alcohol is 20:1-25: 1; the surfactant adopts any one of the following components: sodium dodecyl sulfate, cetyl methyl triamine bromide, polyvinylpyrrolidone and sodium lauroyl ammonia acid; the pH value of the lysate is 4-4.5;
the reagent for the frozen dry powder PCR amplification system is prepared by adopting the following method: mixing a PCR reaction buffer solution, DNA polymerase, a reaction specific primer, a probe and a freeze-drying protective agent solution according to a preset proportion, and then carrying out freeze-drying in three stages; wherein, the first stage is pre-freezing, slowly cooling to-35 to-40 ℃, the cooling time is longer than 1 hour, and keeping for 2.5 hours under the temperature condition; the second stage is a freeze drying stage, the temperature is maintained at-15 to-20 ℃, and the temperature is maintained for 5 hours, wherein the air pressure is maintained at 0.1 mbar; the third stage is enhanced drying, the temperature is kept at 15-25 ℃, the drying time is 2 hours, and the air pressure is kept at 0.1 mbar; wherein, the freeze-drying protective agent adopted for preparing the reagent of the freeze-dried powder PCR amplification system comprises: mannitol, polyethylene glycol, maltose, ascorbic acid and polyvinylpyrrolidone, wherein deionized water added with pyrophosphatase is used for preparing the components to be used as the freeze-drying protective agent solution, wherein the addition amount of mannitol is 3-5g/10 ML; the adding amount of the polyethylene glycol is 2.5-3g/10 ML; the addition amount of maltose is 15-18g/10 ML; the addition amount of ascorbic acid is 0.4-0.6g/10 ML; polyvinylpyrrolidone 0.8-1.5 g/ML; the adding amount of the pyrosulfatase is 1.1-1.6g/10 ML; the reagent for preparing the frozen dry powder PCR amplification system also comprises MoS2, and the adding amount is 55 mug/10 ML; and the PCR reaction buffer solution adopted for preparing the reagent of the frozen dry powder PCR amplification system comprises: deoxyribonucleoside triphosphates, magnesium chloride.
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| CN110016475B (en) * | 2019-05-10 | 2021-07-06 | 宁波艾捷康宁生物科技有限公司 | Excrement DNA extraction kit |
| CN112301097B (en) * | 2019-07-26 | 2022-10-21 | 申翌生物科技(杭州)有限公司 | Sample lysis and PCR reaction composition |
| CN111394346A (en) * | 2020-03-20 | 2020-07-10 | 基蛋生物科技股份有限公司 | Preparation method and application of freeze-dried microspheres of RNA nucleic acid releasing agent and PCR amplification reagent |
| CN111808847A (en) * | 2020-08-12 | 2020-10-23 | 济南国益生物科技有限公司 | Release agent for rapidly extracting nucleic acid by one-step method and preparation and use methods thereof |
| CN114717223A (en) * | 2022-03-29 | 2022-07-08 | 翌圣生物科技(上海)股份有限公司 | Reagent for treating biological sample and its application |
| CN116497085A (en) * | 2022-12-13 | 2023-07-28 | 南京诺唯赞生物科技股份有限公司 | Virus splitting preservation solution |
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| CN107022651B (en) * | 2017-06-02 | 2021-01-29 | 辽宁润基生物科技有限公司 | Kit for rapidly detecting hepatitis C virus nucleic acid and detection method thereof |
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