CN106841608A - A kind of preparation of the quick detection method and its test strips for making a definite diagnosis feline leukemia - Google Patents
A kind of preparation of the quick detection method and its test strips for making a definite diagnosis feline leukemia Download PDFInfo
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- CN106841608A CN106841608A CN201611186051.3A CN201611186051A CN106841608A CN 106841608 A CN106841608 A CN 106841608A CN 201611186051 A CN201611186051 A CN 201611186051A CN 106841608 A CN106841608 A CN 106841608A
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/569—Immunoassay; Biospecific binding assay; Materials therefor for microorganisms, e.g. protozoa, bacteria, viruses
- G01N33/56983—Viruses
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
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- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/531—Production of immunochemical test materials
- G01N33/532—Production of labelled immunochemicals
- G01N33/533—Production of labelled immunochemicals with fluorescent label
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
- G01N2333/005—Assays involving biological materials from specific organisms or of a specific nature from viruses
- G01N2333/08—RNA viruses
- G01N2333/15—Retroviridae, e.g. bovine leukaemia virus, feline leukaemia virus, feline leukaemia virus, human T-cell leukaemia-lymphoma virus
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Abstract
The invention provides a kind of preparation of the quick detection method and its test strips made a definite diagnosis for feline leukemia, the present invention is with feline leukaemia virus (Feline leukemia virus, abbreviation FeLv) it is object, it is prepared for accordingly with chemical covalent coupling method, using the immune chromatography method of fluorescence labeling in feline leukaemia virus detects.Provided by the present invention for the test strips of feline leukaemia virus quick detection, sensitivity is high, feminine gender/positive findings can be drawn while can also quantify the content for providing feline leukaemia virus in sample, test result is relatively sharp and stability is higher, can at room temperature preserve the long period.
Description
Technical field
The present invention relates to a kind of preparation of the quick detection method and its test strips made a definite diagnosis for feline leukemia, belong to doctor
Learn inspection field.
Background technology
The common cause of the worldwide cat infection of feline leukaemia virus (FeLv), and with the incidence of disease higher and death
Rate.The regular incidence disease of feline leukemia has malignant lymphatic tumor, myelomatosis and denaturation property atrophy of thymus gland and irreproducibility
Anaemia etc., wherein to cat most serious being malignant lymphatic tumor.Feline leukaemia virus (FeLv) is characterised by that cat can be triggered
Infection, is not in early days obvious symptom throughout one's life, and the viremia virusemia of continuation occurs therewith, and anaphase is very tired
Difficulty, eventually produces fatal result.Accurately make a definite diagnosis feline leukemia and give in time in morbidity early stage and treat, be to cure the white blood of cat
The optimal selection of disease.The sick cat of persistent infection becomes the direct contagion source of feline leukaemia virus (FeLv), its saliva, excrement
Just, urine, milk, nasal secretion contain virus, by respiratory tract, alimentary infection to Healthy Cats.The sick cat that will be made a definite diagnosis every
From the effective way for being control feline leukaemia virus (FeLv).Therefore, a kind of detection method for quick and precisely making a definite diagnosis feline leukemia
It is the key for controlling and treating feline leukemia.
Feline leukemia early symptom is not obvious, only according to clinical symptoms it is difficult to make a definite diagnosis, to make a definite diagnosis feline leukemia, effectively
Detection method is essential.At present, conventional detection method has：Blood routine examination, pathological section, Enzyme-linked Immunosorbent Assay, immune glue
FeLV in the sick feline tissue of the methods such as body technology for gold detection.Wherein blood routine examination and pathological section detection detection time is more long,
The effect of quick diagnosis is not had.The testing cost of Enzyme-linked Immunosorbent Assay is of a relatively high, and the influence factor in detection process
It is more, easily there are false positive results, for the detection of feline leukemia, the enzyme linked immunosorbent assay degree of accuracy is not good enough.Meanwhile, collaurum
Detection signal is weaker, sensitivity is low limits its application.Therefore a kind of stabilization is needed, it is easy to operate, take short, sensitivity high
Detection method be used for feline leukemia and making a definite diagnosis.
The content of the invention
Easy to operate it is an object of the invention to one kind based on this, time-consuming short, sensitivity is high, quantitative determination feline leukemia disease
The preparation of the method and its test paper of poison, realizes the quick detection of feline leukemia early stage virus.The method is lateral using fluorescent quantitation
Immunochromatography principle detects cat whole blood, blood plasma, serum, the feline leukaemia virus (FeLv) in saliva or tears.It is of the present invention
The fluorescent microsphere of antibody labeling can be a kind of metal colloid particles, a kind of magnetic particle, include but are not limited to latex nanometer
Particle.The antibody of this fluorescent microsphere mark, the preparation method by taking fluorescent latex microballoon as an example is comprised the following steps：
(1) quantitative fluorescent latex microspheres solution is added in cushioning liquid and is diluted, ultrasonic 1.5min obtains fluorescent microsphere
Dispersion liquid.
(2) activator is instilled to rapid in the fluorescent latex microballoon dispersion liquid in step (1), is quickly placed into turbula shaker
After upper low speed concussion 1min, it is fixed on turbula shaker, at room temperature concussion activation 30min.
(3) after activation terminates, by the fluorescent latex microballoon dispersion liquid in step (2) at 26 DEG C, under 14000rpm, centrifugation
Supernatant is removed after 12min, is cleaned with the cushioning liquid in step (1), after being centrifuged with the same terms again, abandoning supernatant,
Redissolved with cushioning liquid in step (4) resuspended.
(4) FeLv monoclonal antibodies are taken, adds cushioning liquid to be diluted to 0.06mg/mL.
(5) nanoparticulate dispersion redissolved in step (3) is added in the antibody diluent in step (4), at once
1.5min is shaken on vortex oscillator, subsequent ultrasound 1.5min is then attached on turbula shaker shake anti-at room temperature
Answer 1h.
(6) after reaction terminates, solution ultrasound 1min in step (5), at 26 DEG C, under 14000rpm, is moved after centrifugation 12min
Supernatant is removed, confining liquid is added, concussion reaction 1h at room temperature is equally then attached on turbula shaker.
(7) confining liquid will be again added to clean after solution centrifugal in step (6), at 26 DEG C, under 14000rpm, centrifugation
Supernatant is removed after 12min, the antibody of fluorescent microsphere mark is can obtain.
The cushioning liquid of step (1) is 2- (N- morpholines) ethyl sulfonic acid cushioning liquid, and its pH is 7.2.
Activator is EDC＆NHS activators in step (2).
The cushioning liquid and the cushioning liquid of step (4) redissolved in step (3) are boric acid acid buffering solution, its pH value
It is 7.8.
Present invention also offers a kind of test strips of the antibody of the fluorescent microsphere mark prepared based on the above method, its feature
It is that the structure of kit is as shown in Figure 1：Sample pad (2) is pasted in plastic bottom board (1) one end, one end of sample pad is closely pressed
The pad (3) of the feline leukaemia virus antibody for being coated with fluorescent microsphere mark is connect, it is fine that pad (3) one end closely crimps nitric acid
Dimension element NC films (4), is coated with detection line T1 (5) and nature controlling line C (6) on NC films, the other end of NC films connects adsorptive pads (7) and formed
Test paper, test paper loads plastics and gets stuck interior formation test card.Including following number of assembling steps：
(1) re-suspension liquid is added in the fluorescent microsphere of antibody labeling, is blown and beaten repeatedly, subsequent ultrasound 5min is until precipitation is completely molten
Solution.
(2) the fluorescent microsphere solution of the antibody labeling in step (1) is sprayed at pad with film metal spraying machine is drawn, is then existed
In baking oven 1h is dried at 37 DEG C.
(3) control line and detection line are wanted into coated antibody, respectively with suitable diluted after, with draw a film metal spraying machine
It is sprayed on nitrocellulose filter respectively, 1h is dried in an oven at 37 DEG C.
(4) pad, sample pad are pasted in assembling, and sealing overnight, is allowed to fully bonding.
The solubility of mark antibody fluorescence microspheres solution is 1.5mg/mL in step (2), and coated weight is 5 μ L/cm.
The coated antibody of control line of nitrocellulose filter is anti-mouse lgG polyclonal antibodies in step (3), and coated weight is
0.9μL/cm；The coated antibody of detection line is FeLv monoclonal antibodies, and coated weight is 0.9 μ L/cm.
The invention provides the application method of this feline leukaemia virus test strip：
(1) pipettor is used, cat serum, blood plasma, whole blood or the saliva sample for taking determined volume are added to sample diluting liquid
In, after being sufficiently mixed, the test fluid for taking determined volume is added in the loading slot of test strips, uses supporting fluorescent quantitative detector
Device, prints examining report after 15min, can draw feminine gender/positive findings while can also quantify and provide feline leukaemia virus in sample
Content.
The invention provides above-mentioned preparation method prepare test strips feline leukemia detection in application.
Relative to prior art, the present invention proposes the detection method method and its test strips of feline leukemia, there is following advantage：
(1) method of antibody labeling of the present invention is using the functional group on fluorescent microsphere surface, using chemistry altogether
The mode that valence link is closed is combined with antibody so that the fluorescent microsphere of antibody labeling is more stable.
(2) when test paper of the present invention is detected for feline leukemia, using supporting fluorescent quantitative detector device, only 15
Accurate result is can obtain in minute, detection time is greatlyd save.
(3) when test paper of the present invention is detected for feline leukemia, can accurately determine compared with collaurum, enzyme linked immunosorbent assay
Measure to amount the amount of feline leukaemia virus (FeLv).
(4) when test paper of the present invention is detected for feline leukemia, LDL can reach 4ng/mL, and sensitivity is higher than
Collaurum, enzyme linked immunosorbent assay.
Brief description of the drawings
Accompanying drawing of the invention is used for providing a further understanding of the present invention, and the illustrated embodiment in the present invention is used to illustrate
The present invention, does not constitute to the present invention and limits.
Fig. 1 is the structural representation of feline leukaemia virus test strip in present invention.
Fig. 2 is the monodispersity the result of the fluorescent microsphere labelled antibody in the embodiment of the present invention one.
Fig. 3 is the fluorescent stability the result of the antibody of the fluorescent microsphere mark in the embodiment of the present invention one.
Specific embodiment
The present invention is described in detail below, but is not limited to the scope of the present invention.
Embodiment one
A kind of fluorescent stability checking of the antibody of fluorescent microsphere mark for feline leukemia detection, including following step
Suddenly：
(1) quantitative fluorescent latex microspheres solution is added in cushioning liquid and is diluted, ultrasonic 1.5min obtains fluorescent microsphere
Dispersion liquid.
(2) activator is instilled to rapid in the fluorescent latex microballoon dispersion liquid in step (1), is quickly placed into turbula shaker
After upper low speed concussion 1min, it is fixed on turbula shaker, at room temperature concussion activation 30min.
(3) after activation terminates, by the fluorescent latex microballoon dispersion liquid in step (2) at 26 DEG C, under 14000rpm, centrifugation
Supernatant is removed after 12min, is cleaned with the cushioning liquid in step (1), after being centrifuged with the same terms again, abandoning supernatant,
Redissolved with cushioning liquid in step (4) resuspended.
(4) FeLv monoclonal antibodies are taken, adds cushioning liquid to be diluted to 0.06mg/mL.
(5) nanoparticulate dispersion redissolved in step (3) is added in the antibody diluent in step (4), at once
1.5min is shaken on vortex oscillator, subsequent ultrasound 1.5min is then attached on turbula shaker shake anti-at room temperature
Answer 1h.
(6) after reaction terminates, solution ultrasound 1min in step (5), at 26 DEG C, under 14000rpm, is moved after centrifugation 12min
Supernatant is removed, confining liquid is added, concussion reaction 1h at room temperature is equally then attached on turbula shaker.
(7) confining liquid will be again added to clean after solution centrifugal in step (6), at 26 DEG C, under 14000rpm, centrifugation
Supernatant is removed after 12min, the fluorescent microsphere of antibody labeling is can obtain.
The cushioning liquid of step (1) is 2- (N- morpholines) ethyl sulfonic acid cushioning liquid, and its pH is 7.2.
Activator is EDC＆NHS activators in step (2).
The cushioning liquid and the cushioning liquid of step (4) redissolved in step (3) are boric acid acid buffering solution, its pH value
It is 7.8.
Present invention also offers a kind of test strips of the antibody of the fluorescent microsphere mark prepared based on the above method, including with
Lower number of assembling steps：
(1) re-suspension liquid is added in the fluorescent microsphere of antibody labeling, is blown and beaten repeatedly, subsequent ultrasound 5min is until precipitation is completely molten
Solution.
(2) the fluorescent microsphere solution of the antibody labeling in step (1) is sprayed at pad with film metal spraying machine is drawn, is then existed
In baking oven 1h is dried at 37 DEG C.
(3) control line and detection line are wanted into coated antibody, respectively with suitable diluted after, with draw a film metal spraying machine
It is sprayed on nitrocellulose filter respectively, 1h is dried in an oven at 37 DEG C.
(4) pad, sample pad are pasted in assembling, and sealing overnight, is allowed to fully bonding.
The solubility of mark antibody fluorescence microspheres solution is 2mg/mL in step (2), and coated weight is 5 μ L/cm.
The coated antibody of control line of nitrocellulose filter is anti-mouse lgG polyclonal antibodies in step (3), and extension rate is
11 times, coated weight is 0.9 μ L/cm；The coated antibody of detection line is FeLv monoclonal antibodies, and extension rate is 10 times, coated weight
It is 0.9 μ L/cm.
Determine the fluorescent value of fluorescent microsphere with supporting fluorescent quantitative detector device in different periods respectively.
Experimental result.
1. the checking of the fluorescent microsphere monodispersity of the antibody labeling prepared by：This fluorescent microsphere dispersion liquid can be at 4 DEG C
Preserve, dispersity stabilization does not find precipitation, as shown in Figure 1 yet after one week.Long-term to stand, the precipitation of appearance only needs pipettor
Suction is beaten for several times or appropriate ultrasound, uses quick single dispersing again by physical shear power.
2. the fluorescent stability of the antibody of the fluorescent microsphere mark for preparing is characterized, as shown in Fig. 2 time change therewith, glimmering
There is no otherness change in the fluorescence total amount of the antibody of light microballoon mark, the antibody that this fluorescent microsphere is marked has good
Fluorescent stability.
Embodiment two
A kind of preparation method of test strips for feline leukaemia virus quick detection, comprises the following steps：
(1) quantitative fluorescent latex microspheres solution is added in cushioning liquid and is diluted, ultrasonic 1.5min obtains fluorescent microsphere
Dispersion liquid.
(2) activator is instilled to rapid in the fluorescent latex microballoon dispersion liquid in step (1), is quickly placed into turbula shaker
After upper low speed concussion 1min, it is fixed on turbula shaker, at room temperature concussion activation 30min.
(3) after activation terminates, by the fluorescent latex microballoon dispersion liquid in step (2) at 26 DEG C, under 14000rpm, centrifugation
Supernatant is removed after 12min, is cleaned with the cushioning liquid in step (1), after being centrifuged with the same terms again, abandoning supernatant,
Redissolved with cushioning liquid in step (4) resuspended.
(4) FeLv monoclonal antibodies are taken, adds cushioning liquid to be diluted to 0.06mg/mL.
(5) nanoparticulate dispersion redissolved in step (3) is added in the antibody diluent in step (4), at once
1.5min is shaken on vortex oscillator, subsequent ultrasound 1.5min is then attached on turbula shaker shake anti-at room temperature
Answer 1h.
(6) after reaction terminates, solution ultrasound 1min in step (5), at 26 DEG C, under 14000rpm, is moved after centrifugation 12min
Supernatant is removed, confining liquid is added, concussion reaction 1h at room temperature is equally then attached on turbula shaker.
(7) confining liquid will be again added to clean after solution centrifugal in step (6), at 26 DEG C, under 14000rpm, centrifugation
Supernatant is removed after 12min, the fluorescent microsphere of antibody labeling is can obtain.
The cushioning liquid of step (1) is 2- (N- morpholines) ethyl sulfonic acid cushioning liquid, and its pH is 7.2.
Activator is EDC＆NHS activators in step (2).
The cushioning liquid and the cushioning liquid of step (4) redissolved in step (3) are boric acid acid buffering solution, its pH value
It is 7.8.
Present invention also offers a kind of test strips of the fluorescent microsphere of the antibody labeling prepared based on the above method, including with
Lower number of assembling steps：
(4) re-suspension liquid is added in the fluorescent microsphere of antibody labeling, is blown and beaten repeatedly, subsequent ultrasound 5min is until precipitation is completely molten
Solution.
(5) the fluorescent microsphere solution of the antibody labeling in step (1) is sprayed at pad with film metal spraying machine is drawn, is then existed
In baking oven 1h is dried at 37 DEG C.
(6) control line and detection line are wanted into coated antibody, respectively with suitable diluted after, with draw a film metal spraying machine
It is sprayed on nitrocellulose filter respectively, 1h is dried in an oven at 37 DEG C.
(7) pad, sample pad are pasted in assembling, and sealing overnight, is allowed to fully bonding.
The solubility of mark antibody fluorescence microspheres solution is 2mg/mL in step (2), and coated weight is 5 μ L/cm.In step (3)
The coated antibody of control line of nitrocellulose filter is anti-mouse lgG polyclonal antibodies, and extension rate is 11 times, and coated weight is 0.9 μ
L/cm；The coated antibody of detection line is FeLv monoclonal antibodies, and extension rate is 10 times, and coated weight is 0.9 μ L/cm.
The invention provides the application method of this feline leukaemia virus test strip：
(1) pipettor is used, cat serum, blood plasma, whole blood or the saliva sample for taking determined volume are added to sample diluting liquid
In, after being sufficiently mixed, the test fluid for taking determined volume is added in the loading slot of test strips, uses supporting fluorescent quantitative detector
Device, prints examining report after 15min, can draw feminine gender/positive findings while can also quantify and provide feline leukaemia virus in sample
Content.
Experiment and the result
1. 50 parts of clinical samples, interval result of determination are determined using the test strips of present case feline leukaemia virus quick detection
It is 98.6% with the coincidence rate of sample value.If Fig. 3 is two groups of negative and positive replicate test results, two groups of test results are complete
Meet clinical effectiveness entirely.The test strips of feline leukaemia virus quick detection of the present invention can delicately quantitative determination sample
In feline leukaemia virus.
2. verified using the stability of the test strips of present case feline leukaemia virus quick detection：Test strips after assembling,
Respectively two weeks, surrounding, take out after eight weeks, take same test sample detection, testing result CV% is relatively low, shows this test strips
With good stability.
Claims (11)
1. a kind of preparation method of the antibody of fluorescent microsphere mark, comprises the following steps：
(1) quantitative fluorescent latex microspheres solution is added in cushioning liquid and is diluted, ultrasonic 1.5min obtains fluorescent microsphere dispersion
Liquid.
(2) activator is instilled to rapid in the fluorescent latex microballoon dispersion liquid in step (1), is quickly placed into low on turbula shaker
After speed concussion 1min, it is fixed on turbula shaker, at room temperature concussion activation 30min.
(3) after activation terminates, by the fluorescent latex microballoon dispersion liquid in step (2) at 26 DEG C, under 14000rpm, 12min is centrifuged
After remove supernatant, cleaned with the cushioning liquid in step (1), again with the same terms be centrifuged after, abandoning supernatant uses step
(4) cushioning liquid redissolves resuspended in.
(4) FeLv monoclonal antibodies are taken, adds cushioning liquid to be diluted to 0.06mg/mL.
(5) nanoparticulate dispersion redissolved in step (3) is added in the antibody diluent in step (4), at once in whirlpool
1.5min is shaken on rotation oscillator, subsequent ultrasound 1.5min is then attached on turbula shaker concussion reaction 1h at room temperature.
(6) after reaction terminates, solution ultrasound 1min in step (5), at 26 DEG C, under 14000rpm, is removed after centrifugation 12min
Clear liquid, adds confining liquid, is equally then attached on turbula shaker concussion reaction 1h at room temperature.
(7) confining liquid will be again added to clean after solution centrifugal in step (6), at 26 DEG C, under 14000rpm, after centrifugation 12min
Supernatant is removed, the antibody of fluorescent microsphere mark is can obtain.
2. the preparation method of the antibody of a kind of fluorescent microsphere mark according to claim 1, it is characterised in that：Step (1)
Cushioning liquid be 2- (N- morpholines) ethyl sulfonic acid cushioning liquid, its pH is 7.2.
3. the preparation method of the antibody for being marked according to a kind of fluorescent microsphere of claim 1, it is characterised in that：Activation in step (2)
Agent is EDC＆NHS activators.
4. the preparation method of the antibody of a kind of fluorescent microsphere mark according to claim 1, it is characterised in that：Step (3)
The cushioning liquid of the cushioning liquid and step (4) of middle redissolution is boric acid acid buffering solution, and its pH value is 7.8.
5. the preparation method of the antibody of a kind of fluorescent microsphere mark according to claim 1, it is characterised in that：Fluorescent microsphere
Can be a kind of metal colloid particles, a kind of magnetic particle, include but are not limited to latex nano particle.
6. preparation method described in a kind of 1~5 any one of usage right requirement prepares kind of the Antibody preparation of fluorescent microsphere mark
Test strips are characterized in that, the structure of kit is as shown in Figure 1：Sample pad (2), sample pad are pasted in plastic bottom board (1) one end
One end closely crimping be coated with fluorescent microsphere mark hair leukemia virus antibody pad (3), pad (3) one end is tight
Close crimping nitrocellulose NC films (4), is coated with detection line T1 (5) and nature controlling line C (6), the other end connection of NC films on NC films
Adsorptive pads (7) form test paper, and test paper loads plastics and gets stuck interior formation test card.
7. the Antibody preparation that the fluorescent microsphere that prepared by preparation method described in a kind of 1~6 any one of usage right requirement is marked
Test strips, comprise the following steps：
(1) re-suspension liquid is added in the fluorescent microsphere of (1) antibody labeling, is blown and beaten repeatedly, subsequent ultrasound 5min is until precipitation is completely molten
Solution.
(2) the fluorescent microsphere solution of the antibody labeling in step (1) is sprayed at pad with film metal spraying machine is drawn, then in baking oven
In dry 1h at 37 DEG C.
(3) control line and detection line are wanted into coated antibody, respectively with suitable diluted after, with draw film metal spraying machine by its
It is sprayed on nitrocellulose filter respectively, 1h is dried in an oven at 37 DEG C.
(4) pad, sample pad are pasted in assembling, and sealing overnight, is allowed to fully bonding.
8. test strips of the Antibody preparation of fluorescent microsphere according to claim 7 mark, it is characterised in that：In step (2)
The solubility of labelled antibody fluorescent microsphere solution is 1.5mg/mL, and coated weight is 5 μ L/cm.
9. test strips of the Antibody preparation of fluorescent microsphere according to claim 6 mark, it is characterised in that：In step (3)
The coated antibody of control line of nitrocellulose filter is anti-mouse lgG polyclonal antibodies, and coated weight is 0.9 μ L/cm；Detection line is coated with
Antibody be FeLv monoclonal antibodies, coated weight be 0.9 μ L/cm.
10. the antibody of the fluorescent microsphere mark that prepared by the preparation method as described in any one of Claims 1 to 5 is in feline leukemia disease
Application in poison detection.
The application of test strips as described in 11. such as claim 6~9 any one in feline leukaemia virus detects.
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Cited By (3)
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| CN108441950A (en) * | 2018-04-28 | 2018-08-24 | 无锡英特派金属制品有限公司 | A kind of preparation method of nano silver wires with high length-diameter ratio |
| CN109991413A (en) * | 2017-12-31 | 2019-07-09 | 江苏雷森生物科技有限公司 | A kind of feline calicivirus fluorogenic quantitative detection card and detection method |
| CN111539932A (en) * | 2020-04-22 | 2020-08-14 | 四川省肿瘤医院 | Hemoglobin measuring instrument and method |
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| CN111539932A (en) * | 2020-04-22 | 2020-08-14 | 四川省肿瘤医院 | Hemoglobin measuring instrument and method |
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