CN111454913A - Novel coronavirus SARS-Cov-2 preserving fluid and preparation method and application thereof - Google Patents
Novel coronavirus SARS-Cov-2 preserving fluid and preparation method and application thereof Download PDFInfo
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Abstract
The invention relates to a novel coronavirus SARS-Cov-2 preservation solution and a preparation method and application thereof, wherein the virus preservation solution 1L comprises the following components, by weight, 0.05-0.4 g of casein, 0.02-0.4 g of a neutralizing antibody, 0.012-0.054 g of gentamicin, 1.5-6 g of bovine serum albumin, 5-20 g of sodium chloride, 3.4-9.8 g of L-glutamic acid and the balance of a buffer solution.
Description
Technical Field
The invention belongs to the field of in vitro diagnosis, and relates to a novel coronavirus SARS-Cov-2 preserving fluid, a preparation method and application thereof.
Background
In order to meet the requirements of epidemic situation prevention and control, improve the convenience, improve the detection accuracy, realize the rapid diagnosis of suspected patients and the field screening of close contact people, the development of rapid detection reagents for nucleic acid, antibody and antigen is needed, and meanwhile, in order to improve the detection accuracy, the post-treatment and the preservation of sample extraction are very important.
Disclosure of Invention
In view of the above, the main objective of the present invention is to provide a novel coronavirus SARS-Cov-2 preservation solution, and a preparation method and an application thereof.
In order to achieve the aim, the invention provides a novel coronavirus SARS-Cov-2 preservation solution, the virus preservation solution of 1L comprises the following components in parts by weight:
the balance being buffer solution.
In a specific embodiment of the present invention, wherein said neutralizing antibody comprises at least one of a pair of anti-SARS antibodies, a pair of anti-syncytial virus antibodies, a pair of anti-adenovirus antibodies, a pair of anti-influenza a virus antibodies and a pair of anti-influenza b virus antibodies.
In a specific embodiment of the present invention, wherein when said neutralizing antibody comprises a pair of anti-SARS antibodies, a pair of anti-syncytial virus antibodies, a pair of anti-adenovirus antibodies, a pair of anti-influenza a virus antibodies and a pair of anti-influenza b virus antibodies, said pair of anti-SARS antibodies: a pair of anti-syncytial virus antibodies: an anti-adenovirus antibody: an anti-influenza a virus antibody: the weight ratio of the first pair of anti-influenza b virus antibodies is 1: (0.5-2): (0.5-2): (0.5-2): (0.5-2).
In a specific embodiment of the present invention, when the neutralizing antibody comprises any two of a pair of anti-SARS antibodies, a pair of anti-syncytial virus antibodies, a pair of anti-adenovirus antibodies, a pair of anti-influenza a virus antibodies, and a pair of anti-influenza b virus antibodies, the ratio of the two is 1: (0.5-2).
In a specific embodiment of the present invention, when the neutralizing antibody comprises any three of a pair of anti-SARS antibodies, a pair of anti-syncytial virus antibodies, a pair of anti-adenovirus antibodies, a pair of anti-influenza a virus antibodies, and a pair of anti-influenza b virus antibodies, the ratio of the three is 1: (0.5-2): (0.5-2).
In a specific embodiment of the present invention, when the neutralizing antibody comprises any four of a pair of anti-SARS antibodies, a pair of anti-syncytial virus antibodies, a pair of anti-adenovirus antibodies, a pair of anti-influenza a virus antibodies, and a pair of anti-influenza b virus antibodies, the ratio of the four is 1: (0.5-2): (0.5-2): (0.5-2).
In a specific embodiment of the present invention, the buffer is one of Tris-HCl buffer, PB buffer, or barbituric sodium-HCl buffer.
In order to achieve the above object, the present invention also provides a method for preparing a novel coronavirus SARS-Cov-2 preservation solution, comprising the steps of: uniformly mixing casein, neutralizing antibody, gentamicin, bovine serum albumin and buffer solution according to the formula amount to obtain the novel coronavirus SARS-Cov-2 preservation solution.
In order to achieve the purpose, the invention also provides a use method of the novel coronavirus SARS-Cov-2 preservation solution, which comprises the following steps of adding 0.2-2 m L virus preservation solution into a plastic hose, a sample cup or a reagent cup, adding a collected sample into the preservation solution, uniformly mixing, and preserving the preservation solution containing the sample at 2-8 ℃ or directly taking 30-100 mu L for immunoassay.
In a specific embodiment of the present invention, wherein the sample is selected from one of a nasal swab, a pharyngeal swab, an oral swab, saliva, sputum or an alveolar lavage.
In a specific embodiment of the present invention, wherein the immunoassay is selected from one of chemiluminescence, enzyme-linked immunosorbent assay, immunochromatography, magnetic particle chemiluminescence, electrochemical immunization and mass-spectrometric immunization.
The invention has the beneficial effects that:
1. the novel coronavirus storage solution provided by the invention has better compatibility with different samples such as a nasal swab, a pharyngeal swab, an oral swab, saliva, sputum, blood and the like, provides a proper and stable buffer environment for a virus sample, and is beneficial to sample detection and storage.
2. The novel coronavirus preservative solution provided by the invention can reduce nonspecific adsorption in the detection process, can neutralize SARS virus, syncytial virus, adenovirus, influenza A virus, influenza B virus and the like in a sample, effectively reduces cross reaction, and improves the detection sensitivity and specificity.
Drawings
FIG. 1 is a schematic diagram of an immunochromatographic test strip for rapidly detecting a novel coronavirus N protein provided by the present invention;
FIG. 2A is a schematic diagram of the internal structure of an upper cover of the immunochromatographic assay card for rapid detection of N protein of a novel coronavirus according to the present invention;
FIG. 2B is a schematic diagram of the internal structure of a bottom groove of the immunochromatography detection card for rapidly detecting the N protein of the novel coronavirus provided by the present invention;
the test strip detection device comprises a PVC plate 1, a coating pad 2, a marker pad 3, a water absorption pad 4, a detection line 5, a quality control line 6, a marker joint 7, a sample 8, a sample pad 9, a top cover 11, a bottom groove 12, a sample adding hole 13, an observation window 14, a test strip placement area 15, a positioning column 16, a positioning hole 17, a first limiting part 18, a second limiting part 19 and a third limiting part 20.
Detailed Description
The invention is further described with reference to the following specific embodiments and the accompanying drawings.
The following materials or reagents, unless otherwise specified, are commercially available.
The preservation solution of the novel coronavirus SARS-Cov-2 can also be referred to as a virus preservation solution for short.
EXAMPLE 1 preparation of a novel coronavirus SARS-Cov-2 virus stock solution
The formula comprises 200mg of casein, 30mg of gentamicin, 2000mg of bovine serum albumin, 9g of NaCl, 4.8g of L-glutamic acid;
one anti-SARS antibody 10mg, the other anti-SARS antibody 10 mg;
10mg of anti-syncytium virus antibody and 10mg of anti-syncytium virus antibody of the other strain;
10mg of anti-adenovirus antibody on one strain and 10mg of anti-adenovirus antibody on the other strain;
10mg of anti-influenza A virus antibody and 10mg of anti-influenza A virus antibody of the other strain;
10mg of anti-influenza B virus antibody and 10mg of anti-influenza B virus antibody of the other strain;
the raw materials are subjected to constant volume of 1L by using 0.02M PB buffer solution, mixed uniformly and kept stand for later use.
The 0.02M PB buffer solution was prepared as follows (1L):
0.2M Na2HPO4the solution was prepared as follows:
0.2M NaH2PO4the solution was prepared as follows:
EXAMPLE 2 preparation of novel coronavirus N protein immunochromatography detection kit
1) Fluorescent microsphere labeling another strain of mouse anti-novel coronavirus N protein monoclonal antibody
Adding 0.5M L fluorescent microspheres into 1mg carbodiimide (EDC) and 1mg N-hydroxysuccinimide (NHS), stirring at room temperature and 120r/min for 3h, adding 100 mu L of another mouse anti-novel coronavirus N protein monoclonal antibody, stirring at room temperature and 120r/min for 1h, adding 10mg BSA confining liquid, stirring at 120r/min for 1h, centrifuging at 2-8 ℃ and 12000 r/min for 20min, removing supernatant, re-dissolving the solid precipitate obtained after centrifugation into 1M L by using 0.2M phosphate buffer (pH 7.4), adding 1 mu L Proclin300, and storing at 4 ℃ for later use.
2) Labeling rabbit IgG polyclonal antibody by fluorescent microsphere
Adding 1mg of carbodiimide (EDC) and 1mg of N-hydroxysuccinimide (NHS) into 0.5M L fluorescent microspheres, stirring at room temperature and 120r/min for 3h, then adding 100 mu L mg of goat rabbit IgG polyclonal antibody, stirring at room temperature and 120r/min for 1h, then adding 10mg of BSA confining liquid, continuing stirring at 120r/min for 1h, centrifuging at 2-8 ℃ and 11000r/min for 30 min, removing supernatant, finally re-dissolving the solid precipitate to 1M L by using 0.2M phosphate buffer (pH 7.4), then adding 1 mu L Proclin300, and storing at 4 ℃ for later use.
3) Preparation of coating pad
A mouse anti-novel coronavirus N protein monoclonal antibody and a goat anti-rabbit polyclonal antibody are respectively diluted to 1mg/M L by 0.2M phosphate buffer solution (pH 7.4), membrane scribing is carried out on a nitrocellulose membrane (NC membrane) by a membrane scribing and gold spraying instrument, a detection line (T line) 5 containing the mouse anti-novel coronavirus N protein monoclonal antibody and a quality control line (C line) 6 containing the goat anti-rabbit polyclonal antibody are used, and then the coating pad 2 is prepared by drying for 4 hours at 37 ℃ and under the environment of the humidity of less than 30%.
4) Preparation of marker pad
① the glass cellulose membrane was soaked in 0.2M phosphate buffer (pH 7.4) for 6h and then dried at 35 ℃ for 8 h.
② A mouse anti-novel coronavirus N protein monoclonal antibody marked by fluorescent microspheres with a molar ratio of 1:1 and fluorescent latex microspheres marked by fluorescent microsphere marked rabbit IgG are uniformly mixed, sprayed on a glass cellulose membrane at a speed of 10 mu L/cm, and then placed at 45 ℃ for drying for 2 hours to prepare a marker pad 3, and a place, which is coated with the mouse anti-novel coronavirus N protein monoclonal antibody marked by the fluorescent microspheres and the fluorescent microsphere marked rabbit IgG, on the marker pad 3 is called a marker binding part 7.
5) Assembly of immunochromatography detection kit
Firstly, a nitrocellulose membrane 2 is bonded on a PVC plate 1, then a water absorption pad 4 is lapped at one end close to a quality control line 6 on the nitrocellulose membrane 2, a marker pad 3 and a sample pad 9 connected with the marker pad are lapped at one end close to a detection line 5 of the nitrocellulose membrane 2, a strip cutting machine is used for cutting the test strip (shown in figure 1) into 4mm +/-0.1 mm, and the test strip is put into a card shell to prepare the novel coronavirus N protein immunochromatography detection kit.
The card housing is selected from the prior art, for example, the card housing (as shown in fig. 2) may include: a bottom tank 12 connected to the PVC plate 1; an upper cover 11 connected to the bottom tank 12, the upper cover 11 being provided with a sample application hole 13 for applying a sample to the sample pad 9; and the observation window 14 is arranged on the upper cover 11 and is used for data acquisition of the detection line 5 and the quality control line 6.
As shown in fig. 2B, the bottom tank 12 includes: a plurality of positioning holes 17 which are symmetrically distributed and positioned on the inner surface of the test strip, wherein a plurality of first limiting parts 18 used for limiting the test strip to move transversely and second limiting parts 19 used for limiting the test strip to move longitudinally are arranged among the plurality of positioning holes 17; the first limiting part 18 and the second limiting part 19 which are symmetrically arranged enclose a paper strip placing area 15 (a dotted line area) for placing the test paper strip;
as shown in fig. 2A, the upper cover 11 includes: a plurality of positioning posts 16 which are matched with a plurality of positioning holes 17, and are matched with each other to fix the upper cover 11 and the bottom groove 12 together; the upper cover 11 further includes a third limiting portion 20 for limiting the up and down movement of the test strip.
An observation window 14 for data acquisition is arranged above the coated pad 2 to expose all the detection lines 5 and the quality control lines 6 for collecting detection results; and the observation window 14 is arranged on the upper cover 11 at a position corresponding to the middle part of the test strip placement area 15. The upper cover 11 is provided with a sample adding hole at a position corresponding to the sample pad 9 for dropwise adding the sample 8 on the sample pad 9. The distance between the detection line and the sample adding hole is 15-25 mm.
Example 3 clinical assay use
Adding 0.5m L of the virus preservation solution prepared in the embodiment 1 into a plastic hose, immersing a cotton rod after collecting throat swab and nose swab samples in the virus preservation solution and stirring, extruding the outer side of the plastic hose for multiple times by fingers to enable the virus preservation solution to fully soak the cotton rod, then pulling out the cotton rod, twisting out (namely mixing the sample collected on the cotton rod into the virus preservation solution) to obtain a sample to be detected, detecting the sample to be detected by adopting the novel coronavirus N protein immunochromatographic kit described in the embodiment 2, dripping the sample to be detected into a sample adding hole of the kit, standing for 15min, inserting the kit into a fluorescence immunochromatographic analyzer for detection, automatically calculating the T/C value of the sample by the analyzer, and judging whether the sample is positive or negative according to the normal range of the sample T/C value, or when two fluorescence bands appear under ultraviolet lamp irradiation, obtaining the detection result which is shown in the following table 1.
TABLE 1
As can be seen from the clinical detection data in Table 1, the detection results of the pharyngeal swab samples detected by the novel coronavirus N-protein immunochromatographic kit comprise 9 samples of patients with novel coronavirus pneumonia, 7 samples of the patients with novel coronavirus pneumonia are positive, and the positive detection rate is 77%; all samples of 2 patients with influenza A virus were tested negative; all the samples of 3 patients with influenza B virus are detected as negative; all 10 normal human samples were tested negative. In the detection results of the nasal swab samples detected by adopting the novel coronavirus N protein immunochromatographic kit, 6 samples of 9 novel coronavirus pneumonia patient samples are detected to be positive, and the positive detection rate is 77%; all samples of 2 patients with influenza A virus were tested negative; all the samples of 3 patients with influenza B virus are detected as negative; all 10 normal human samples were tested negative.
EXAMPLE 3 SARS Cross-reaction validation
Virus preservation solution 1: the neutralizing antibody is not added, and the preparation method comprises the following steps:
virus preservation solution 2: adding an anti-SARS antibody, the concrete preparation is as follows:
preparing SARS recombinant protein solution with concentration of 5 mug/m L by sample preservative fluid 1 as control sample;
preparing SARS recombinant protein solution with concentration of 5 mug/m L by sample preservation solution 2 as detection sample;
then, 60 μ L of the control sample and the test sample were separately taken and tested by the novel coronavirus N-protein immunochromatographic kit, and the results are shown in Table 2 below.
TABLE 2
| Detection value (T/C) | Normal value range (T/C) | |
| Control sample | 0.062 | 0-0.05 |
| Test sample | 0.009 | 0-0.05 |
As can be seen from the results of the tests in Table 2, the addition of a pair of anti-SARS antibodies to the sample storage solution significantly reduced the SARS cross-reaction.
The above-mentioned embodiments are merely examples provided to fully illustrate the present invention, and the scope of the present invention is not limited thereto. The equivalent substitution or change made by the technical personnel in the technical field on the basis of the invention is all within the protection scope of the invention. The protection scope of the invention is subject to the claims.
Claims (11)
1. A novel coronavirus SARS-Cov-2 preservation solution is characterized in that the 1L virus preservation solution comprises the following components in parts by weight:
2. the novel preservative solution for coronavirus SARS-Cov-2 as claimed in claim 1, wherein the neutralizing antibody comprises at least one of a pair of anti-SARS antibodies, a pair of anti-syncytial virus antibodies, a pair of anti-adenovirus antibodies, a pair of anti-influenza a virus antibodies, and a pair of anti-influenza b virus antibodies.
3. The novel preservative solution for coronavirus SARS-Cov-2 as claimed in claim 2, wherein when the neutralizing antibody comprises a pair of anti-SARS antibodies, a pair of anti-syncytial virus antibodies, a pair of anti-adenovirus antibodies, a pair of anti-influenza a virus antibodies, and a pair of anti-influenza b virus antibodies, the pair of anti-SARS antibodies: a pair of anti-syncytial virus antibodies: an anti-adenovirus antibody: an anti-influenza a virus antibody: the weight ratio of the anti-influenza B virus antibody to the anti-influenza B virus antibody is 1 to (0.5-2).
4. The novel preservative solution for coronavirus SARS-Cov-2 as claimed in claim 2, wherein when the neutralizing antibody comprises any two of a pair of anti-SARS antibodies, a pair of anti-syncytial virus antibodies, a pair of anti-adenovirus antibodies, a pair of anti-influenza A virus antibodies and a pair of anti-influenza B virus antibodies, the ratio of the two is 1: 0.5-2.
5. The preservation solution for coronavirus SARS-Cov-2 as claimed in claim 2, wherein when the neutralizing antibody comprises any three of a pair of anti-SARS antibodies, a pair of anti-syncytial virus antibodies, a pair of anti-adenovirus antibodies, a pair of anti-influenza A virus antibodies and a pair of anti-influenza B virus antibodies, the ratio of the three is 1: 0.5-2.
6. The novel preservative solution for coronavirus SARS-Cov-2 as set forth in claim 2, wherein when the neutralizing antibody comprises any four of a pair of anti-SARS antibodies, a pair of anti-syncytial virus antibodies, a pair of anti-adenovirus antibodies, a pair of anti-influenza A virus antibodies and a pair of anti-influenza B virus antibodies, the ratio of the four is 1: 0.5-2.
7. The preservation solution for the novel coronavirus SARS-Cov-2 as claimed in claim 1, wherein the buffer is one of Tris-HCl buffer, PB buffer or barbituric sodium-HCl buffer.
8. A method for preparing the novel preservation solution for coronavirus SARS-Cov-2 as described in any one of claims 1 to 7, which comprises the steps of: uniformly mixing casein, neutralizing antibody, gentamicin, bovine serum albumin and buffer solution according to the formula amount to obtain the novel coronavirus SARS-Cov-2 preservation solution.
9. A method of using the novel preservative solution for coronavirus SARS-Cov-2 as described in any one of claims 1 to 7, which comprises the steps of adding a 0.2 to 2m L virus preservative solution into a plastic tube, a sample cup or a reagent cup, adding a collected sample into the preservative solution, mixing the solutions uniformly, and preserving the sample-containing preservative solution at 2 to 8 ℃ or directly taking 30 to 100 μ L for immunoassay.
10. The method of use of claim 9, wherein the sample is selected from one of a nasal swab, a pharyngeal swab, an oral swab, saliva, sputum, or an alveolar lavage.
11. The method of use of claim 9, wherein said immunoassay is selected from the group consisting of chemiluminescence, enzyme-linked immunosorbent assay, immunochromatography, magnetic particle chemiluminescence, electrochemical immunization, and mass-spectrometric immunization.
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Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111808827A (en) * | 2020-08-12 | 2020-10-23 | 泰州鑫联诚润生物技术有限公司 | Virus preserving fluid capable of effectively preserving samples such as viruses for long time |
| CN112662731A (en) * | 2020-12-01 | 2021-04-16 | 杭州金域医学检验所有限公司 | Coronavirus sample preservation solution and application thereof |
| WO2022029083A1 (en) * | 2020-08-06 | 2022-02-10 | Ng Biotech | System for rapid analysis of a biological sample, intended for detecting the presence of at least one analyte in the biological sample |
| WO2022040523A1 (en) * | 2020-08-20 | 2022-02-24 | BiopolariX Scientific, LLC | Saliva testing kit using nano carbon immunochromatography |
| FR3115110A1 (en) * | 2021-02-15 | 2022-04-15 | Biospeedia | IMMUNOLOGICAL TEST FOR DETECTION OF VIRAL INFECTIONS |
| WO2022077592A1 (en) * | 2020-10-15 | 2022-04-21 | 广州达博生物制品有限公司 | Virus preservation reagent |
| CN114934023A (en) * | 2022-06-08 | 2022-08-23 | 万绵水 | Saliva preservation solution and preparation method and application thereof |
| CN115184344A (en) * | 2022-01-30 | 2022-10-14 | 北京市疾病预防控制中心 | Novel coronavirus IgG antibody detection method suitable for novel biological sample oral fluid and application thereof |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20160238601A1 (en) * | 2013-10-14 | 2016-08-18 | The University Of North Carolina At Chapel Hill | Methods and compositions for coronavirus diagnostics and therapeutics |
| CN111398583A (en) * | 2020-02-13 | 2020-07-10 | 北京华科泰生物技术股份有限公司 | Kit for detecting novel coronavirus N protein and application thereof |
-
2020
- 2020-03-17 CN CN202010189261.8A patent/CN111454913A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20160238601A1 (en) * | 2013-10-14 | 2016-08-18 | The University Of North Carolina At Chapel Hill | Methods and compositions for coronavirus diagnostics and therapeutics |
| CN111398583A (en) * | 2020-02-13 | 2020-07-10 | 北京华科泰生物技术股份有限公司 | Kit for detecting novel coronavirus N protein and application thereof |
Non-Patent Citations (5)
| Title |
|---|
| AHMED SF等: "Preliminary Identification of Potential Vaccine Targets for the COVID-19 Coronavirus (SARS-CoV-2) Based on SARS-CoV Immunological Studies", 《VIRUSES-BASEL》 * |
| JASPER FUK-WOO CHAN等: "Genomic characterization of the 2019 novel human-pathogenic coronavirus isolated from a patient with atypical pneumonia after visiting Wuhan", 《EMERG MICROBES INFECT》 * |
| TIAN XL等: "Potent binding of 2019 novel coronavirus spike protein by a SARS coronavirus-specific human monoclonal antibody", 《EMERGING MICROBES & INFECTIONS》 * |
| ZHOU P等: "A pneumonia outbreak associated with a new coronavirus of probable bat origin", 《NATURE》 * |
| 佚名: "https://www.chemicalbook.com/ProductList.aspx?kwd=SARS%E6%8A%97%E4%BD%93&hp=true¤t=hp", 《CHEMICALBOOK》 * |
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