CN101750495A - Magnetic immunochromatographic test strip for detecting hepatitis B core antibody and preparation method thereof - Google Patents
Magnetic immunochromatographic test strip for detecting hepatitis B core antibody and preparation method thereof Download PDFInfo
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- CN101750495A CN101750495A CN 200910236332 CN200910236332A CN101750495A CN 101750495 A CN101750495 A CN 101750495A CN 200910236332 CN200910236332 CN 200910236332 CN 200910236332 A CN200910236332 A CN 200910236332A CN 101750495 A CN101750495 A CN 101750495A
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Abstract
The invention relates to a magnetic immunochromatographic test strip for detecting hepatitis B core antibody in blood and a preparation method thereof. The magnetic immunochromatographic test strip introduces the magnetic immunochromatographic technology and a fluorescein isothiocyanate system into the hepatitis B core antibody detection. The test strip is assembled by sequentially mutually sticking a coating film, a magnetic particle pad combined with the hepatitis B core antibody, a sample pad and a water-absorbing pad on a substrate in a staggered manner and then covering a transparent plastic sealing film on the upper layer. The magnetic immunochromatographic test strip has the advantages of simple operation, fast reaction time, high sensitivity, good specificity and the like.
Description
Technical field
The invention belongs to field of medical examination, particularly relate to a kind of magnetic immuno-chromatographic test paper strip that detects hepatitis B core antibody and preparation method thereof.
Background technology
Hepatitis B is caused that by hepatitis type B virus (HBV) HBV is a kind of genomic enveloped virus of partially double stranded cyclic DNA that comprises, and belongs to Hepadnaviridae.When virus is duplicated in liver cell, can disturb the function of liver, immune system activation produces a series of special reactions with antagonism and elimination infectiousness factor immediately.As a kind of result of pathologic damage, liver is inflamed.The HBV that continues infects the serious pathology result who causes and comprises: chronic liver function is incomplete, cirrhosis and hepatocellular carcinoma (HCC).
Hepatitis B core antibody is to infect the sign antibody that the back occurs as far back as HBV, the height of tiring, longer duration, even do not disappear all the life, so be the good sign of epidemiology survey.It is generally acknowledged that high titre core antibody prompting HBV is duplicating, HBV was previously once infected in the positive expression of low titre.
Present HBcAb diagnostic techniques mainly comprises enzyme linked immune assay (ELISA), chemiluminescence (CLIA), immunochromatographic method (collaurum or latex particle method), PCR method, and these methods all have characteristics and applicable object separately.
The ELISA method is generally to use detection technique in the present clinical blood examination, but ELISA test operation program complexity occur false positive or false negative result easily, and sensitivity is lower.CLIA and ELISA method are similar, just sensitivity has improved some, the problem that the not basic ELISA of solution exists, and all there is complex operation in the two, the problem that reaction time is long, and all need microplate reader or light-emitting appearance and wash complex apparatus such as plate machine and incubator, and can not single part detect, further limited their application in some basic hospitals, clinic.
Having occurred both at home and abroad in recent years with collaurum or latex particle is the quick detection test paper bar of representative, but because the result is the naked eyes visualizations, is subjected to the influence of observer's subjective judgement easily, and sensitivity is low, and result precision is not high yet, and the detection window phase is longer.
PCR method, need very high experiment work environment and condition (between sterile working, superclean bench) and a large amount of instrument and equipment (incubators, hydro-extractor, PCR instrument etc.), and operation is extremely complicated, need be subjected to the personnel operation of strict professional training, and the test period is very long, is not suitable for clinical detection and uses.
(Mgnetic ImmunoChromatographic Test MICT) is a kind of single part of fast quantification detection technique that occurs in recent years to magnetic immuno-chromatographic.Be to replace traditional label (collaurum, latex particle etc.) to carry out immunochromatography, be combined in biochemical substances on the super-paramagnetism nano particulate by detection detection by quantitative data to biological sample are provided with supperparamagnetic particles (superPMPs).This technology is compared with conventional art has following advantages: a. used magnetic detecting instrument adopts the solid phase element, and miniaturized design is had a style of one's own, independent operating, and volume is little, and is easy and simple to handle; B. the high 10-100 of all kinds of range estimation quick diagnosis of remolding sensitivity method doubly; C. linear range is linear in 4 order of magnitude concentration ranges; D. the super-paramagnetism nano particulate can not decayed in time by polymer coating.It is easy fast that this technology has been inherited traditional immunochromatographic method (collaurum, latex particle etc.), the advantage of single part of operation, and it is low to have remedied traditional immunochromatography technique sensitivity again, can only be qualitative, shortcoming that can not be quantitative.
Mark magnetic particle commonly used at present is a super paramagnetic particle (superPMPs), do not have any magnetic in the absence of externally-applied magnetic field, only just can show magnetic adding under the action of a magnetic field, the super paramagnetic particle of commercialization all passes through finishing, greatly facilitate labeling process, mark is easy, good reproducibility.
The fluorescein isothiocynate system is introduced in the magnetic immuno-chromatographic, and the sensitivity that had both improved detection method provides a kind of current techique platform again, has increased the versatility of technology, has reduced the influence of system factor.
Summary of the invention
The objective of the invention is in magnetic immuno-chromatographic technology and the system combined HBcAb of the being applied in immunoassay of fluorescein isothiocynate.To resist FITC antibody covalent coupling on super paramagnetic particle, FITCization HBcAb is mixed with it afterwards as detecting moving phase, the HBcAg bag is made detection line as catching solid phase on nitrocellulose filter, carry out the detection of sample according to routine immunization chromatography ratio juris, detect in conjunction with simple and easy to do magnetism detector, when realizing high-sensitivity detection, can reach the purpose of fast detecting again, can avoid the deficiency of aforementioned several detection methods, combine the advantage of aforementioned several method again: can single part detect, also can batch detection, and can provide quantitative result, surveying instrument is simple and reliable, easy and simple to handle, convenient and practical.
For reaching above-mentioned purpose, technical scheme of the present invention is as follows: the present invention is with coated film, sticks on the base plate in conjunction with the interlaced successively 2mm of magnetic mat of particles, sample pad and the adsorptive pads of HBcAb, cover the transparent plastic diaphragm seal then on the upper strata and make, be coated with the HBcAb detection line on the wherein said coated film in advance, and the nature controlling line of FITCization bovine serum albumin(BSA).
The base plate of selecting for use is the transparent plastic base plate, and coated film is the nitrocellulose filter of 35mm width, and the adsorptive pads of selecting for use is a cellulose membrane, and the magnetic mat of particles is a fiberglass packing, and sample pad is the pretreated cellulose membrane of process sample pad treating fluid.Described sample pad treating fluid is the polyvinyl alcohol (PVA) (PVP) that contains 1%-5% gelatin and 0.1%-1%, and the 0.02M of 0.01-0.2% Tween-20 (Tween-20), the Tris salt buffer of pH7.0-7.6.
Detect the preparation method of the magnetic immuno-chromatographic test paper strip of hepatitis B core antibody, may further comprise the steps:
The preparation of A, magnetic particle: selecting diameter for use is the super paramagnetic particle of 100-300nm, use the mode of carbodiimide (EDC) and succinimide (NHS) covalent coupling will resist the FITC antibody labeling to the magnetic particle, again FITC is attached on the HBcAb, then with 1: 5-1: 20 ratio (volume ratio) is mixed FITCization HBcAb with anti-FITC antibody magnetic particle, and guarantees that anti-FITC antibody magnetic particle is excessive;
B, use quantitative liquid-jet device to be sprayed on the magnetic mat of particles magnetic particle for preparing with the amount of 25 μ l/cm-50 μ l/cm;
The preparation of C, coated film: use bag to be cushioned the concentration that liquid is diluted to HBcAg and FITCization BSA 0.5-2mg/m respectively, use quantitative liquid-jet device respectively with both being interval on the nitrocellulose filter with 0.5-1.0cm, dry the back and in confining liquid, soak after 10 minutes, add drying agent and seal up for safekeeping standby in 25-35 ℃ of oven dry 8 hours;
The processing of D, sample pad: sample pad is put into sample pad Treatment Solution immersion treatment take out 25-35 ℃ of oven dry 8 hours after 1 hour;
The assembling of E, test strips: interlaced successively 2mm sticks coated film, magnetic mat of particles, sample pad, adsorptive pads on the transparent plastic base plate, covers the transparent plastic diaphragm seal then on the upper strata, obtains test paper plate, and the width cutting promptly obtains test strips as requested.
Described steps A comprises following three steps:
1) preparation of anti-FITC antibody magnetic particle: the sodium-acetate buffer washing magnetic particle that uses the 50mM pH4.5-5.0 that contains 0.2%Tween-20, adding EDC and NHS makes the two final concentration be 20mmol, room temperature reaction 1 hour, use contains the 50mM of 0.1%Tween-20, the sodium-acetate buffer of pH4.5-5.0 fully washs and adds anti-FITC antibody behind the magnetic particle to make the molecule ratio of anti-FITC antibody and magnetic particle be 5: 1 (mol ratio), room temperature reaction 3 hours, add the 0.02M that contains 0.5%BSA, the PBS room temperature sealing of pH7.0-7.6 30 minutes, washing magnetic particle, use contains 1%PVP, 1% gelatin, 1%Tween-20, the boric acid of the 50mM pH8.2-9.0 of 5% sucrose preserve damping fluid redissolution magnetic particle, and 4 ℃ of preservations are standby;
2) preparation of FITC-HBcAb: with HBcAb to 0.02M, 4 ℃ of dialysed overnight of the PBS of pH7.0-7.6, adjustment concentration is 2mg/ml, FITC is used dimethyl sulfoxide (DMSO) (DMSO) dissolving, final concentration is 50mM, adds the FITC solution of aequum, room temperature reaction 1 hour with 20: 1 molecule ratios in antibody-solutions, to 4 ℃ of dialysed overnight of 0.02M PBS ,-20 ℃ of preservations are standby behind the adding equal-volume glycerine;
3) FITC-HBcAb and anti-FITC antibody magnetic particle mixes, amount with 0.5 μ l/mg magnetic particle adds FITC-HBcAb in anti-FITC antibody magnetic particle solution, fully use behind the mixing and preserve damping fluid with 1: 5-1: 20 ratio (volume ratio) is used mixture diluted fiberglass packing to be sprayed.
Among the described step B, the spraying method of magnetic particle is: use quantitative spray film device evenly to be sprayed on the glass fibre with the amount of 50 μ l/cm the magnetic particle for preparing, add drying agent after the freeze drying and seal up for safekeeping standby.
Among the described step C, the preparation method of coated film is: be cushioned liquid (0.05M CB with bag, pH9.4-9.8) the HBcAg dilution is 0.5mg/ml, FITCization BSA dilution is 1mg/ml, use quantitative spray film device with the amount of 1 μ l/cm with both with the interval spray printing of 0.6cm on nitrocellulose filter, room temperature is dried after 30 minutes and to be soaked in confining liquid after 10 minutes in 25-35 ℃ of oven dry 8 hours, adds drying agent and seals up for safekeeping standby.
Compare with existing quick detection test paper bar, the present invention uses magnetism detector to carry out result's interpretation, carries out yin and yang attribute according to the magnetic detection value ratio of detection line and nature controlling line and judges, has reduced subjectivity, the result accurately, reliable.
Description of drawings
Fig. 1 detects the magnetic immuno-chromatographic test paper strip structural representation of hepatitis B core antibody for the present invention.
Detect magnetic immuno-chromatographic test paper strip of hepatitis B core antibody and preparation method thereof for further specifying the present invention, describe especially exemplified by following embodiment, this embodiment is in order to explain rather than limit by any way the present invention.
Embodiment
The magnetic immuno-chromatographic test paper strip of hepatitis B core antibody in the detection blood of the present invention, as shown in Figure 1, this test strips is at base plate 1) on interlaced successively 2mm ground paste to go up coated film 2), combine the magnetic mat of particles 3 of HBcAb), sample pad 4), adsorptive pads 5), and cover transparent plastic diaphragm seal 6 on the upper strata) test strips that assembles, coated film 2) on be coated with HBcAb detection line T and nature controlling line C in advance.
In specific embodiment, the HBcAg that adopts, HBcAb are the commercialization antigen-antibody.Utilize the competition law principle to detect the sample of HBcAb; when containing HBcAb in the sample to be measured; antibody on this antibody and the magnetic particle is along with the carrying out of chromatography effect; move forward and arrive the HBcAg bag by line T place; HBcAg competition on both meetings of T place and coated film forms antigen-antibody complex; in addition, unconjugated anti-FITC antibody labeling magnetic particle can continue to move ahead when arriving nature controlling line C, thereby FITCization BSA can combine at C line place with anti-FITC antibody labeling magnetic particle and occurs the magnetic particle aggregation equally.Entire reaction was carried out in 30 minutes fully, general reaction can be used magnetic immuno-chromatographic instrument Card Reader after 15 minutes, T line and C line all can produce corresponding magnetic signal value, calculate the ratio of T/C, get final product the yin and yang attribute of result of determination according to default boundary ratio.Whole Card Reader, calculating, with the process sequencing fully of preset bounds value comparison, magnetism detector can directly provide the yin and yang attribute result.
The preparation method of the magnetic immuno-chromatographic test paper strip of hepatitis B core antibody sees following example in the detection blood of the present invention:
Embodiment 1
Detect the magnetic immuno-chromatographic test paper strip of hepatitis B core antibody in the blood and the preparation method of paper box
The test strips of present embodiment and the preparation method of paper box may further comprise the steps:
The preparation of A, antigen-antibody: select commercial HBcAg, HBcAb for use, to 20mM, the PBS of pH7.2 (pH7.0-7.6 all is suitable for), 4 ℃ of dialysed overnight are standby.
The preparation of B, coated film:
Bag is cushioned the preparation of liquid: the carbonic acid buffer of 50mM pH 9.6 (CB) is cushioned liquid for bag, and the rearmounted 4 ℃ of preservations of 0.22 μ m filtering with microporous membrane degerming are standby, two weeks of the term of validity.
The preparation of confining liquid: contain the phosphate buffer (PBS) of the 20mM pH7.2 (pH7.0-7.6 all is suitable for) of 0.5%BSA, it is standby that 0.22 μ m filtering with microporous membrane degerming is placed on 4 ℃ of preservations, one week of the term of validity.
The preparation of coated film: the CB that is cushioned liquid 50mM pH9.6 with bag is 0.5mg/ml with the HBcAg dilution, FITCization BSA dilution is 1mg/ml, use quantitative spray film device with the amount of 1 μ l/cm with both with the even spray printing in the interval of 0.6cm on 3.5cm width nitrocellulose filter, room temperature is dried after 30 minutes in the confining liquid (PBS that contains the 0.02MpH7.2 (pH7.0-7.6 all is suitable for) of 0.5%BSA,) in 25-35 ℃ of oven dry 8 hours, the adding drying agent was sealed up for safekeeping standby after 10 minutes in middle immersion.
The preparation of C, magnetic particle:
The preparation of sodium-acetate buffer: with distilled water and sodium acetate and glacial acetic acid secure ph is 4.7 (pH4.5-5.0 all is suitable for), concentration is the acetate buffer solution of 50mM, adding Tween-20 is that 4 ℃ of preservations are standby after 0.1%, the 0.22 μ m filtering with microporous membrane degerming to final concentration, two weeks of the term of validity.
Boric acid is preserved the preparation of damping fluid: use distilled water, boric acid and borax preparation pH are 8.5 (pH8.2-9.0 all is suitable for), and final concentration is the borate buffer of 50mM, add PVP, Casine, Tween-20, sucrose, final concentration is respectively 1%, 1%, 0.5%, 5%, 0.22 4 ℃ of preservations are standby after the degerming of μ m filtering with microporous membrane, one week of the term of validity.
The preparation of anti-FITC antibody magnetic particle: use 50mM pH4.7 (pH4.5-5.0 all is suitable for) the sodium-acetate buffer washing magnetic particle that contains 0.1%Tween-20, adding EDC and NHS makes the two final concentration be 20mmol, room temperature reaction 1 hour, fully adding anti-FITC antibody behind the washing magnetic particle, to make the molecule ratio of anti-FITC antibody and magnetic particle be 5: 1 (mol ratio), room temperature reaction 3 hours, the PBS that adds the 20mM pH7.2 (pH7.0-7.6 all is suitable for) that contains 0.5%BSA, room temperature sealing 30 minutes, washing magnetic particle, use contains 1%PVP, 1%Casein, 0.5%Tween-20, the boric acid of the 50mmolpH8.5 of 5% sucrose (pH8.2-9.0 all is suitable for) is preserved damping fluid redissolution magnetic particle, and 4 ℃ of preservations are standby.
The preparation of FITC-HBcAb: with HBcAb 4 ℃ of dialysed overnight of PBS to 20mM pH7.2 (pH7.0-7.6 all is suitable for), adjustment concentration is 2mg/ml, FITC is used the DMSO dissolving, final concentration is 50mM, the FITC solution that in antibody-solutions, adds aequum with 20: 1 molecule ratios, room temperature reaction 1 hour, to 4 ℃ of dialysed overnight of PBS of 0.02M pH7.2 (pH7.0-7.6 all is suitable for) ,-20 ℃ of preservations are standby behind the adding equal-volume glycerine.
FITC-HBcAb mixes with anti-FITC antibody magnetic particle: the amount with 0.5 μ l/mg adds FITC-HBcAb in anti-FITC antibody magnetic particle solution, fully uses behind the mixing and preserves damping fluid with ratio mixing use in 1: 10.
The spraying of D, magnetic particle and freeze-drying
That uses BioDot spray film instrument nozzle specially usedly evenly is sprayed at the magnetic particle handled well the amount with 50 μ l/cm on the 0.8cm width fiberglass packing, the frozen overnight drying, add drying agent seal up for safekeeping standby,
The processing of E, sample pad
1.8cm width sample pad is put into sample pad Treatment Solution immersion treatment take out 25-35 ℃ of oven dry 8 hours after 1 hour.
The sample pad treating fluid is the polyvinyl alcohol (PVA) (PVP) that contains 1%-5% gelatin and 0.1%-1%, and the 0.02M of 0.01-0.2% Tween-20 (Tween-20), the Tris salt buffer of pH7.0-7.6
The assembling of F, test strips and cutting
Following all operations all must carry out in temperature 20-25 ℃ the room in humidity less than 20%.
The assembling of test paper plate: use as requested that 3.5cm is the wide coated film of BioDot LM5000 type assembling instrument, 2.5cm wide thieving paper, the magnetic mat of particles that 0.8cm is wide, the wide sample pad of 1.8cm are assembled on the 9.8em width transparent plastic base plate, stick upper strata transparent plastic cover plate, be assembled into test paper plate.
Cutting of test strips: use BioDot CM4000 type cutting cutter that the test paper plate that assembles is cut into the wide finished product test strips of 0.5cm.
The assembling of G, test card
The single part test strips of well cutting of the present invention is placed in the draw-in groove on the plastic bottom card, covers loam cake, use card press machine up and down two plastic clips compress, guarantee that whole test strips is in tensioned state.Adding the drying agent room temperature seals up for safekeeping standby.
H, determine the 2 D code information of this batch
The name of an article: HBcAb magnetic detection card
Batch: on the assembling date of test card, form is: Year/Month/Day, XXXX/XX/XX
Determining of yin and yang attribute interpretation standard: get 100 parts confirm HBcAb samples (power all has), 500 parts at random sample use this batch test card to detect, use the magnetism detector testing result, calculate the T1/C value of each test card, the T2/C value, use statistical method computation of mean values and standard deviation, determine: T/C＜0.8 is positive, and T/C＞1.0 are negative, is gray area between the two.
The printing of I, two-dimension code is pasted
With in the above-mentioned 2 D code information input two-dimension code printer and print, two-dimension code is pasted on the ad-hoc location of test card, use two-dimension code paste position detecting device to inspect 2% at random by random samples and guarantee that two-dimension code pastes errorless.
J, finished product packing
The single part test card and that the posts two-dimension code drying prescription of being responsible for a task until it is completed is sealed in the aluminium foil bag, 100 person-portions are that a packing places in the packing box, and a instructions of a box and 1 bottle of 10ml dress chromatography damping fluid are promptly made paper box, this paper box keeps in Dark Place in room temperature, and the shelf-life is 18 months.
The chromatography buffer formulation is: 1%Tween-20,0.5% glycerine, 1%NP-40,0.05%NaN3, the PBS of 20mmolpH7.2 (pH7.0-7.6 all is suitable for).
In the blend step except FITC-HBcAb and anti-FITC antibody magnetic particle: fully use behind the mixing preserve damping fluid with 1: 5 ratio with mixture diluted, other step is with embodiment 1.
In the blend step except FITC-HBcAb and anti-FITC antibody magnetic particle: fully use behind the mixing preserve damping fluid with 1: 20 ratio with mixture diluted, other step is with embodiment 1.
Embodiment 4
The using method of test card of the present invention
1, application of sample
From packing box, take out single part test card, tear the aluminium foil strip packing, test card is placed on the smooth desktop, get 50 μ l sample serum with micropipettor and add in the well on the card, add 50 μ l chromatography damping fluids again, wait question response to carry out 15 minutes.
2, measurement and result output
The MICT detector is started shooting in advance, test card is inserted the card inserting mouth of detector, the operation instrument, instrument can read the 2 D code information on the card automatically and measure, and prints measurement result immediately, and the yin and yang attribute result can show in print result.
Claims (9)
1. magnetic immuno-chromatographic test paper strip that detects hepatitis B core antibody, it is characterized in that: this test strips is that coated film, the magnetic mat of particles that combines hepatitis B core antibody, sample pad, adsorptive pads are sticked on the base plate successively interlaced 2mm, cover the transparent plastic diaphragm seal then on the upper strata and assemble, be coated with hepatitis B core antibody detection line and nature controlling line on the wherein said coated film in advance.
2. the magnetic immuno-chromatographic test paper strip of detection hepatitis B core antibody according to claim 1, it is characterized in that: the CB that the preparation of described coated film is to use 0.05mM pH9.4-9.8 is diluted to hepatitis B virus core antigen and fluorescein isothiocynate bovine serum albumin(BSA) the concentration of 0.5-2mg/ml respectively, use quantitative liquid-jet device respectively with the two with the interval spray printing of 0.5-1.0cm on nitrocellulose filter, dry the back and in confining liquid, soak after 10 minutes, add drying agent and seal up for safekeeping standby in 25-35 ℃ of oven dry 8 hours.
3. the magnetic immuno-chromatographic test paper strip of detection hepatitis B core antibody according to claim 1, it is characterized in that: described magnetic mat of particles preparation is to select suitable magnetic particle for use, uses the mode of carbodiimide and succinimide covalent coupling will resist the fluorescein isothiocynate antibody labeling to the magnetic particle; Fluorescein isothiocynate is connected on the hepatitis B core antibody, and mixes with anti-fluorescein isothiocynate antibody magnetic particle, guarantee that anti-fluorescein isothiocynate antibody magnetic particle is excessive with proper proportion; The magnetic particle for preparing is sprayed on the glass fibre with the amount of quantitative liquid-jet device with 25 μ l/cm-50 μ l/cm.
4. the magnetic immuno-chromatographic test paper strip of detection hepatitis B core antibody according to claim 1 is characterized in that: described sample pad is through the pretreated cellulose membrane of sample pad treating fluid.
5. the preparation method of coated film according to claim 2, it is characterized in that: described hepatitis B virus core antigen dilute concentration is 1.0mg/ml, fluorescein isothiocynate BSA dilute concentration is 1mg/ml, and both spray printings are spaced apart 0.8cm on nitrocellulose filter.
6. the spraying method of magnetic particle according to claim 3 is characterized in that: described magnetic particle spray coating is 50 μ l/cm in the amount of glass fibre.
7. suitable magnetic particle according to claim 3 is characterized in that: described magnetic particle is that diameter is the super paramagnetic particle of 100-300nm.
8. the volume ratio of fluorescein isothiocynate antibody according to claim 3 and anti-fluorescein isothiocynate antibody magnetic is characterized in that described suitable volumes ratio is 1: 5-1: 20 ratio.
9. sample pad treating fluid according to claim 4, it is characterized in that, described sample pad treating fluid contains the polyvinyl alcohol (PVA) (PVP) of 1%-5% gelatin and 0.1%-1%, and the 0.02M of 0.01-0.2% Tween-20 (Tween-20), the Tris salt buffer of pH7.0-7.6.
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102183649A (en) * | 2011-01-31 | 2011-09-14 | 中国农业大学 | Bovine tuberculosis antigen specific gamma-interferon enzyme-linked immuno sorbent assay (ELISA) kit |
| CN103293295A (en) * | 2013-05-07 | 2013-09-11 | 上海爱纳玛斯医药科技有限公司 | Magnetic biological probe and test strip for detecting hepatitis B virus (HBV) and preparation method and using method of biological probe |
| CN104597255A (en) * | 2015-02-04 | 2015-05-06 | 华中农业大学 | Test paper card for detecting pseudorabies virus gE protein antibody in porcine serum as well as preparation method and application thereof |
| CN109298189A (en) * | 2018-11-15 | 2019-02-01 | 迈克生物股份有限公司 | HCG detection reagent, test paper and its application method |
-
2009
- 2009-10-16 CN CN 200910236332 patent/CN101750495A/en active Pending
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102183649A (en) * | 2011-01-31 | 2011-09-14 | 中国农业大学 | Bovine tuberculosis antigen specific gamma-interferon enzyme-linked immuno sorbent assay (ELISA) kit |
| CN103293295A (en) * | 2013-05-07 | 2013-09-11 | 上海爱纳玛斯医药科技有限公司 | Magnetic biological probe and test strip for detecting hepatitis B virus (HBV) and preparation method and using method of biological probe |
| CN104597255A (en) * | 2015-02-04 | 2015-05-06 | 华中农业大学 | Test paper card for detecting pseudorabies virus gE protein antibody in porcine serum as well as preparation method and application thereof |
| CN109298189A (en) * | 2018-11-15 | 2019-02-01 | 迈克生物股份有限公司 | HCG detection reagent, test paper and its application method |
| CN109298189B (en) * | 2018-11-15 | 2021-08-06 | 迈克生物股份有限公司 | HCG detection reagent, test paper and use method thereof |
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