CN117887807A - Freeze-drying protective agent capable of improving full premix performance of RNA detection reagent and application thereof - Google Patents
Freeze-drying protective agent capable of improving full premix performance of RNA detection reagent and application thereof Download PDFInfo
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- 238000001514 detection method Methods 0.000 title claims abstract description 43
- 239000003153 chemical reaction reagent Substances 0.000 title claims abstract description 36
- 238000004108 freeze drying Methods 0.000 title abstract description 50
- 239000003223 protective agent Substances 0.000 title abstract description 36
- ZHNUHDYFZUAESO-UHFFFAOYSA-N Formamide Chemical compound NC=O ZHNUHDYFZUAESO-UHFFFAOYSA-N 0.000 claims abstract description 16
- 239000002736 nonionic surfactant Substances 0.000 claims abstract description 8
- 244000052769 pathogen Species 0.000 claims description 14
- 230000001717 pathogenic effect Effects 0.000 claims description 14
- 238000002360 preparation method Methods 0.000 claims description 8
- HDTRYLNUVZCQOY-UHFFFAOYSA-N α-D-glucopyranosyl-α-D-glucopyranoside Natural products OC1C(O)C(O)C(CO)OC1OC1C(O)C(O)C(O)C(CO)O1 HDTRYLNUVZCQOY-UHFFFAOYSA-N 0.000 claims description 5
- CTKXFMQHOOWWEB-UHFFFAOYSA-N Ethylene oxide/propylene oxide copolymer Chemical group CCCOC(C)COCCO CTKXFMQHOOWWEB-UHFFFAOYSA-N 0.000 claims description 5
- HDTRYLNUVZCQOY-WSWWMNSNSA-N Trehalose Natural products O[C@@H]1[C@@H](O)[C@@H](O)[C@@H](CO)O[C@@H]1O[C@@H]1[C@H](O)[C@@H](O)[C@@H](O)[C@@H](CO)O1 HDTRYLNUVZCQOY-WSWWMNSNSA-N 0.000 claims description 5
- 229920001993 poloxamer 188 Polymers 0.000 claims description 5
- 229940044519 poloxamer 188 Drugs 0.000 claims description 5
- 125000000647 trehalose group Chemical group 0.000 claims description 2
- 238000003860 storage Methods 0.000 abstract description 15
- 239000012295 chemical reaction liquid Substances 0.000 abstract description 11
- 238000012360 testing method Methods 0.000 abstract description 5
- 238000004519 manufacturing process Methods 0.000 abstract description 4
- 238000012216 screening Methods 0.000 abstract description 4
- 230000008859 change Effects 0.000 abstract description 3
- 239000007788 liquid Substances 0.000 description 13
- 102000004190 Enzymes Human genes 0.000 description 11
- 108090000790 Enzymes Proteins 0.000 description 11
- 241000193990 Streptococcus sp. 'group B' Species 0.000 description 11
- 238000000034 method Methods 0.000 description 11
- 239000008188 pellet Substances 0.000 description 9
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 6
- 238000007792 addition Methods 0.000 description 6
- 238000003752 polymerase chain reaction Methods 0.000 description 6
- 230000008569 process Effects 0.000 description 6
- 239000000243 solution Substances 0.000 description 6
- 238000006243 chemical reaction Methods 0.000 description 5
- 230000008901 benefit Effects 0.000 description 4
- 230000009467 reduction Effects 0.000 description 4
- 239000000523 sample Substances 0.000 description 4
- HDTRYLNUVZCQOY-LIZSDCNHSA-N alpha,alpha-trehalose Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@@H]1O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 HDTRYLNUVZCQOY-LIZSDCNHSA-N 0.000 description 3
- 230000003321 amplification Effects 0.000 description 3
- 238000001035 drying Methods 0.000 description 3
- 230000000694 effects Effects 0.000 description 3
- 238000005516 engineering process Methods 0.000 description 3
- 239000000203 mixture Substances 0.000 description 3
- 229910052757 nitrogen Inorganic materials 0.000 description 3
- 238000003199 nucleic acid amplification method Methods 0.000 description 3
- 102000004169 proteins and genes Human genes 0.000 description 3
- 108090000623 proteins and genes Proteins 0.000 description 3
- 230000035945 sensitivity Effects 0.000 description 3
- 239000000126 substance Substances 0.000 description 3
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 3
- 241000894006 Bacteria Species 0.000 description 2
- 230000001580 bacterial effect Effects 0.000 description 2
- 239000011324 bead Substances 0.000 description 2
- 230000018044 dehydration Effects 0.000 description 2
- 238000006297 dehydration reaction Methods 0.000 description 2
- 238000007710 freezing Methods 0.000 description 2
- 230000008014 freezing Effects 0.000 description 2
- 239000000463 material Substances 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 102000039446 nucleic acids Human genes 0.000 description 2
- 108020004707 nucleic acids Proteins 0.000 description 2
- 150000007523 nucleic acids Chemical class 0.000 description 2
- 238000011056 performance test Methods 0.000 description 2
- 230000037452 priming Effects 0.000 description 2
- 230000001737 promoting effect Effects 0.000 description 2
- 239000012085 test solution Substances 0.000 description 2
- 239000012807 PCR reagent Substances 0.000 description 1
- 229930182555 Penicillin Natural products 0.000 description 1
- JGSARLDLIJGVTE-MBNYWOFBSA-N Penicillin G Chemical compound N([C@H]1[C@H]2SC([C@@H](N2C1=O)C(O)=O)(C)C)C(=O)CC1=CC=CC=C1 JGSARLDLIJGVTE-MBNYWOFBSA-N 0.000 description 1
- 239000013543 active substance Substances 0.000 description 1
- 239000000654 additive Substances 0.000 description 1
- 230000000996 additive effect Effects 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 238000001816 cooling Methods 0.000 description 1
- HAAZLUGHYHWQIW-KVQBGUIXSA-N dGTP Chemical class C1=NC=2C(=O)NC(N)=NC=2N1[C@H]1C[C@H](O)[C@@H](COP(O)(=O)OP(O)(=O)OP(O)(O)=O)O1 HAAZLUGHYHWQIW-KVQBGUIXSA-N 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 238000004925 denaturation Methods 0.000 description 1
- 230000036425 denaturation Effects 0.000 description 1
- 238000003745 diagnosis Methods 0.000 description 1
- 150000002016 disaccharides Chemical class 0.000 description 1
- 238000004090 dissolution Methods 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 238000009472 formulation Methods 0.000 description 1
- 230000004927 fusion Effects 0.000 description 1
- 208000015181 infectious disease Diseases 0.000 description 1
- 150000002500 ions Chemical class 0.000 description 1
- 239000012931 lyophilized formulation Substances 0.000 description 1
- 238000002844 melting Methods 0.000 description 1
- 230000008018 melting Effects 0.000 description 1
- 230000000813 microbial effect Effects 0.000 description 1
- 239000004005 microsphere Substances 0.000 description 1
- 229940049954 penicillin Drugs 0.000 description 1
- 239000002994 raw material Substances 0.000 description 1
- 238000003757 reverse transcription PCR Methods 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 238000013112 stability test Methods 0.000 description 1
- 230000000087 stabilizing effect Effects 0.000 description 1
- 238000010257 thawing Methods 0.000 description 1
- 238000009777 vacuum freeze-drying Methods 0.000 description 1
- 244000052613 viral pathogen Species 0.000 description 1
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/68—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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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
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Abstract
The invention discloses a freeze-drying protective agent capable of improving full premix performance of an RNA detection reagent and application thereof, and the freeze-drying protective agent comprises non-reducing sugar, formamide, 7-deaza-dGTP and a nonionic surfactant. Aiming at the technical problems that the fully premixed reaction liquid is unstable and the freeze-drying protective agent affects the performance and the appearance of the detection reagent in the production of the RNA freeze-drying detection reagent, the invention provides the freeze-drying protective agent which can promote the fully premixed stability of the RNA detection reagent, does not affect the performance of the reaction liquid and does not affect the storage performance of the freeze-drying appearance. Through screening test of a plurality of protective agents, a group of protective agents which have no influence on the RNA detection reagent and have no obvious change of performance in 72 hours are screened out, and the protective agents with good appearance performance are stored at 45 ℃ for 90 days after freeze-drying.
Description
Technical Field
The invention belongs to the technical field of biology, and relates to a freeze-drying protective agent, in particular to a freeze-drying protective agent capable of improving full premix performance of a pathogen RNA detection reagent.
Background
The pathogen nucleic acid fluorescence PCR detection method has the advantages of rapidness, accuracy, high sensitivity and the like. However, since the DNA structure is stable, it can remain positive for a long time after bacterial death, and the specific stage of pathogen infection cannot be judged, thereby causing excessive treatment. The RNA has short half-life period, can be rapidly degraded after bacterial death, can be used for separately detecting pathogen RNA, can distinguish the activity of bacteria, and has higher sensitivity and accuracy compared with the PCR (polymerase chain reaction) and other technologies which take DNA as a target due to the fact that RNA has multiple copies in non-viral pathogen microbial cells. RNA not only has the advantages of high sensitivity of nucleic acid detection, but also can screen dead bacteria, provides more accurate detection, and is more suitable for clinical screening and diagnosis.
RT enzyme in the RNA detection reagent is sensitive, is easily influenced by temperature and repeated freeze thawing, the cost of reagent transportation and storage is high, a plurality of enzymes exist in the RNA reagent, the addition is complicated during detection, and the freeze drying technology can be used for directly freeze-drying MIX containing the enzyme, storing and transporting the MIX at normal temperature, and the freeze-dried product can be directly re-dissolved by using a sample, so that the detection efficiency of the product can be improved to a certain extent.
The freeze-drying technique is a technique in which a substance containing a large amount of water is frozen into ice by cooling in advance, and then the ice is sublimated under vacuum to achieve the drying purpose. The freeze-drying process is carried out under the conditions of low temperature and vacuum, so that biological, chemical or physical changes of heat-sensitive substances are effectively inhibited, and active substances in the raw materials are preserved. However, various stresses existing in the freeze-dried product during the whole process, including low temperature stress, freezing stress, drying stress and the like, are often factors directly or indirectly causing protein denaturation in the product, so that a protective agent is required to be used in the freeze-drying process. To obtain an ideal lyophilized product, a suitable lyoprotectant is important, which will affect the subsequent performance and storage stability for the PCR reagents.
The lyoprotectant on the market may have the problems of reduced performance of the reaction solution of the RNA detection solution, poor stability and poor storage stability after lyophilization, for example:
1. in the preparation process of the freeze-dried ball, after the preparation of the reaction liquid is finished, a bead dropping machine is required to prepare the ice ball, and due to the fact that the production yield is high, the bead dropping machine has limited productivity, and a period of time is required to be reserved between the full premixing of the reagent and the preparation of the ice ball, so that the full premixing of the reagent is required to be stable within a certain period of time. The RT-PCR detection reagent contains RT enzyme, taq enzyme and buffer containing various ions, and especially the RT enzyme is sensitive and unstable in a full premix state.
2. Screening tests are carried out by using enzymes of different manufacturers, and the phenomenon of 72h performance reduction of full premix storage at 4 ℃ can occur.
3. The reason of the performance reduction after full premixing is inquired, and the phenomenon of premix signal reduction is found when RT enzyme and priming detection exist simultaneously, and when RT enzyme or priming detection is not added in the reagent, the signals are not different from the existing preparation after premixing storage. Therefore, it is needed to find a freeze-drying protective agent, which can prevent the combination of primer and RT enzyme at low temperature to realize full premix stability without affecting the freeze-drying appearance and performance.
Disclosure of Invention
In order to overcome the defects of the prior art, the freeze-drying protective agent capable of improving the full-premixing performance of the pathogen RNA detection reagent provided by the invention can stabilize the full-premixing reaction liquid at 4 ℃ for 72 hours on the basis of not influencing the reaction liquid, and can form good appearance after freeze-drying without influencing the performance.
In order to achieve the above purpose, the present invention adopts the following technical scheme:
the invention provides a lyoprotectant capable of improving full premix performance of a pathogen RNA detection reagent, which comprises non-reducing sugar, formamide, 7-deaza-dGTP and a nonionic surfactant.
As a preferred embodiment of the present invention, the non-reducing sugar is trehalose.
As a preferred embodiment of the present invention, the nonionic surfactant is poloxamer 188.
As a preferable scheme of the invention, the freeze-drying protective agent comprises the following components in parts by weight: 3-6 parts of non-reducing sugar, 0.1-0.4 part of formamide, 1-3 parts of 7-deaza-dGTP and 0.5-1 part of non-ionic surfactant.
The invention also provides application of the freeze-drying protective agent in preparation of pathogen RNA detection reagents.
Aiming at the technical problems that the full premix reaction liquid is unstable, the freeze-drying protective agent influences the performance and the appearance of the detection reagent and the like in the production of the RNA freeze-drying detection reagent, the invention provides the freeze-drying protective agent which can promote the full premix stability of the RNA detection reagent, does not influence the performance of the reaction liquid and does not influence the storage performance of the freeze-drying appearance. Through screening test of a plurality of protective agents, a group of protective agents which have no influence on the RNA detection reagent and have no obvious change of performance in 72 hours are screened out, and the protective agents with good appearance performance are stored at 45 ℃ for 90 days after freeze-drying.
Compared with the prior art, the invention has the following beneficial effects:
1) The freeze-drying protective agent has strong compatibility, stability and convenient transportation, and improves the detection limit.
2) The freeze-drying protective agent has the advantages of promoting the stability of the fully premixed reaction liquid, not affecting the detection performance of the reagent, being freeze-dried and having no influence on the performance after freeze-drying.
Drawings
FIG. 1 is a schematic representation of a lyophilized pellet prepared according to the present invention.
Fig. 2 is a graph of a liquid unprotected group versus a group a protectant test.
Fig. 3 is a graph comparing liquid non-protectant group with group F protectant tests.
Detailed Description
In order to facilitate understanding of the technical means, the creation characteristics, the achievement of the objects and the effects achieved by the present invention, the present invention is further described below with reference to specific examples, but the following examples are only preferred examples of the present invention, not all of which are described in detail below. Based on the examples in the embodiments, those skilled in the art can obtain other examples without making any inventive effort, which fall within the scope of the invention. The experimental methods in the following examples are conventional methods unless otherwise specified, and materials, reagents, etc. used in the following examples are commercially available unless otherwise specified.
The invention provides a freeze-drying protective agent for promoting full premixing stability of an RNA detection reagent, which does not influence the performance of a reaction solution and the storage performance of freeze-drying appearance, and mainly comprises 4 components: 1 part of trehalose is 3-6 parts of trehalose is natural sugar, is non-reducing sugar composed of special disaccharide molecules, can serve as a low-temperature protective agent in the freezing process, can also serve as a dehydration protective agent in the drying and dehydration process, and can reduce protein browning reaction of biological products to deteriorate and deactivate. 2 formamide (DMF) 0.1-0.4 parts formamide is a PCR additive, can promote DNA melting to prevent primer probe combination in PCR reaction, and plays an important role in full premix stability. 3 7-deaza-dGTP 1-3 parts, 7-deaza-dGTP is a dGTP analogue which can modify the template to avoid secondary structure during PCR reaction. In the full premix performance detection of the RNA detection reagent, when RT enzyme and primer are fully premixed for 72 hours in a system, the amplification performance is obviously reduced, and the condition of the performance reduction can be improved by adding 7-deaza-dGTP during the full premix. 4 poloxamer 188 0.5-1 part poloxamer 188 is a common high molecular pharmaceutical auxiliary material, is a nonionic surfactant, and also plays a certain role in protecting protein in the freeze drying process.
Example 1
The embodiment provides a freeze-drying protective agent of an RNA detection reagent, which comprises trehalose, formamide, 7-deaza-dGTP and poloxamer 188, wherein the formula of the freeze-drying protective agent is shown in table 1.
TABLE 1 lyoprotectant formulation
Example 2:
effect of the lyoprotectants of the groups obtained in example 1 on the performance of Group B Streptococcus (GBS) RNA detection solutions
(1) Primer and probe sequences for group B streptococcus and internal standard (see Table 2)
Primer and probe sequences for group B streptococcus and internal standard
(2) Preparation of group B streptococcus RNA detection liquid
The pathogen group B streptococcus detection system (25. Mu.L) provided in this example is specifically as shown in Table 3:
TABLE 3 detection System
The protective agent A, B, C, D, E, F, G of example 1 was added to each group
(3) The liquid properties and the properties after 72 hours of storage at 4℃are shown in Table 4.
TABLE 4 Performance test
Results: the addition of the protective agent A, B, C, D, F, G in the GBS RNA detection liquid has no obvious influence on the liquid performance, and the performance of the protective agent E is reduced; after the reagent is stored for 72 hours at the temperature of 4 ℃, the performance of the group without the addition of the protective agent and the performance of the group with the addition of the B, E, F, G protective agent are slightly reduced, which shows that the reagent is unstable in full premix storage at the temperature of 4 ℃, and the addition of the A, C, D protective agent has a certain stabilizing effect on the full premix reaction liquid.
Example 3:
this example provides preparation of lyophilized spheres of group B Streptococcus RNA detection solution
(1) Microsphere formation
Pouring liquid nitrogen into a sterilized container, respectively dripping the reaction liquid (A group-F group) prepared in the embodiment 1 into the liquid nitrogen by using a liquid separation system, condensing into round pellets by 25 mu l each, waiting for 10 seconds until the pellets sink into the bottom of the liquid nitrogen, and respectively fishing up the pellets and transferring the pellets into the pre-cooled corresponding penicillin bottles.
(2) Vacuum freeze drying
After the reagent pellets were collected, the pellets were transferred to a lyophilizer and the lyophilization procedure is shown in table 5:
TABLE 5 lyophilization procedure
(3) Appearance referring to fig. 1, it can be seen that groups a-F each formed a good appearance.
Example 4
This example provides the lyophilized pellet properties of group B Streptococcus RNA test solutions obtained in example 3
(1) Appearance and reconstitution time are shown in table 6.
TABLE 6 appearance and reconstitution time
Results: the group 7 group B streptococcus RNA freeze-dried balls keep spherical shape and have smooth surfaces, no adhesion or fusion phenomenon, good re-solubility and instant dissolution after being added with water.
(2) Moisture content of lyophilized formulation
The moisture content of the freeze-dried pellets was measured by a card moisture meter and is shown in table 7.
TABLE 7 moisture content
Results: except the group C, the water content of the other groups is lower than 3%, and the index has great influence on the storage stability of the freeze-dried balls
(3) Post lyophilization performance test
After rehydration of the lyophilized product, GBS RNA was detected and the amplification procedure is shown in table 8.
TABLE 8 amplification procedure
The criteria are shown in Table 9.
TABLE 9 criterion for judging
The results of the judgment are shown in Table 10.
TABLE 10 judgment results
Liquid unprotected VS a group protectant (after lyophilization) see fig. 2, liquid unprotected VS F group protectant (after lyophilization) see fig. 3.
Results: the performance of the freeze-dried A, B, C and E is not changed obviously, and the performance of the freeze-dried D, F and G is reduced.
(4) Freeze-dried storage stability test, results are shown in table 11.
Table 11.45 ℃ test solutions performance after 3 months storage
| Experimental grouping | Ct(FAM) | Ct(HBB) |
| Liquid non-protective agent | 17.53 | 13.12 |
| Group A protectant (45 ℃ C. 3 months after freeze-drying) | 18.68 | 12.95 |
| Group B protectant (45 ℃ C. 3 months after freeze-drying) | 19.21 | 13.29 |
| Group C protectant (45 ℃ C. 3 months after freeze-drying) | 27.32 | 15.11 |
| Group D protectant (45 ℃ C. 3 months after freeze-drying) | 40 | 20.28 |
| Group E protectant (45 ℃ C. 3 months after freeze-drying) | 26.62 | 17.52 |
| Group F protectant (45 ℃ C. 3 months after freeze-drying) | 29.68 | 19.44 |
| Group G protectant (45 ℃ C. 3 months after freeze-drying) | 40 | 21.78 |
Results: the self-made protectant group A B had no significant change in performance after 3 months of storage.
To sum up: after the group A protective agent is added, the appearance and the performance are stable after freeze-drying and storage, and the performance is stable within 72 hours after full premixing of the reaction liquid before freeze-drying, so that the production requirement is met.
While the invention has been described with respect to preferred embodiments thereof, it will be understood by those skilled in the art that various modifications and additions may be made without departing from the scope of the invention. Equivalent embodiments of the present invention will be apparent to those skilled in the art having the benefit of the teachings disclosed herein, when considered in the light of the foregoing disclosure, and without departing from the spirit and scope of the invention; meanwhile, any equivalent changes, modifications and evolution of the above embodiments according to the essential technology of the present invention still fall within the scope of the technical solution of the present invention.
Claims (5)
1. The lyoprotectant capable of improving the full premix performance of the pathogen RNA detection reagent is characterized by comprising non-reducing sugar, formamide, 7-deaza-dGTP and a nonionic surfactant.
2. The lyoprotectant capable of improving the full premix performance of a pathogen RNA detection reagent of claim 1, wherein the non-reducing sugar is trehalose.
3. The lyoprotectant capable of improving the full premix performance of a pathogen RNA detection reagent of claim 1, wherein the nonionic surfactant is poloxamer 188.
4. The lyoprotectant capable of improving the full premix performance of a pathogen RNA detection reagent according to any one of claims 1-3, wherein the lyoprotectant comprises the following components in parts by weight: 3-6 parts of non-reducing sugar, 0.1-0.4 part of formamide, 1-3 parts of 7-deaza-dGTP and 0.5-1 part of non-ionic surfactant.
5. The use of a lyoprotectant according to any one of claims 1-4 for improving the full premix performance of a pathogen RNA detection reagent, wherein the lyoprotectant is used in the preparation of a pathogen RNA detection reagent.
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| CN202311793530.1A CN117887807A (en) | 2023-12-25 | 2023-12-25 | Freeze-drying protective agent capable of improving full premix performance of RNA detection reagent and application thereof |
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