CN103308699A - Colloidal gold reagent plate for quickly detecting methyltestosterone in aquatic product - Google Patents
Colloidal gold reagent plate for quickly detecting methyltestosterone in aquatic product Download PDFInfo
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- CN103308699A CN103308699A CN2013102533129A CN201310253312A CN103308699A CN 103308699 A CN103308699 A CN 103308699A CN 2013102533129 A CN2013102533129 A CN 2013102533129A CN 201310253312 A CN201310253312 A CN 201310253312A CN 103308699 A CN103308699 A CN 103308699A
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Abstract
The invention relates to a colloidal gold reagent plate for quickly detecting methyltestosterone in an aquatic product, which can be used for quickly detecting methyltestosterone in the aquatic product. The reagent plate provided by the invention consists of an upper plastic formwork, a lower plastic formwork and a backing, as well as a sample cushion, a colloidal gold combination cushion, a nitrocellulose film and a water absorbing cushion, which are sequentially and tightly adhered to the backing. The nitrocellulose film is sequentially sprayed with a detecting wire and a quality control wire along the direction from the sample cushion to the water absorbing cushion, and a test result of the methyltestosterone can be determined by the macroscopic color. After the colloidal gold reagent plate is used, the semi-quantitative visual detection can be carried out, the whole operation process can be carried out only for 15 minutes without the help of any expensive experimental facilities, the large-scale sample selection can be preferably carried out, and the colloidal gold reagent plate is applicable to large-scale quick detection of the methyltestosterone in the aquatic product in the business sector, the inspection and quarantine institution, the aquiculture enterprises, and the processing enterprises.
Description
Technical field
The present invention relates to a kind of colloidal gold reagent plate of fast detecting Methyltestosterone, be specifically related to the immune colloid gold quick detection reagent plate of Methyltestosterone in a kind of fast detecting aquatic products.
Background technology
Methyltestosterone (methyltestosterone, MT) is a kind of anabolic steroids, belongs to protein anabolic hormone, has male and the protein assimilating double action.But its residual serious threat health in aquatic products.Great many of experiments shows, the methyltestosterone metabolism time is long, is chronic, at a specified future date and cumulative to the mankind's harm.Its harm is mainly manifested in the balance of disturbing natural hormone in the body, normal person's physiological function is got muddled, and also can cause neonate's deformity when serious, affect children's normal growth growth.
The 95/22/EC of European Union provides against and uses the male sex hormone veterinary drug, the U.S. stipulates that also the maximum residue limit(MRL) (MRL) of its similar testosterone propionate in muscle is 0.64 μ g/kg, China calendar year 2001 the industry standard " fishing medicine residue limits in the pollution-free food aquatic products " implemented by the Ministry of Agriculture (NY5070-2001) in, classify methyltestosterone as forbidden drugs.The regulation methyltestosterone must not detect in all Edible tissues of all edible animals for banning use of veterinary drug in No. 235 bulletins of the Ministry of Agriculture of issue in 2002 " animal food herbal medicine maximum residue limit(MRL) ".
The hormone residues analytical approach mainly contains high performance liquid chromatography (HPLC), liquid chromatography-mass spectrography determination method (LC-MS) etc. in the at present domestic and international aquatic products, its characteristics are that accuracy is high, highly sensitive, good reproducibility, yet, these methods need long experimental period and the experimental facilities of corresponding costliness, be difficult to satisfy that modern measure is quick, simple and direct, the requirement of scene, be unsuitable for extensive sample examination.
Colloidal gold immunity chromatography is the immunology detection technology of a kind of simple and fast of setting up on the basis of immunity percolation technology, whole or the major part that the method will be reacted needed raw material all has been incorporated in the reagent, reaction only needs several minutes to tens of minutes usually, and testing result is brought interpretation with macroscopic colour developing bar.This method has easy quick, simple to operate, high specificity, does not need the advantages such as extras.
Summary of the invention
The objective of the invention is to overcome the defective that existing detection technique exists, develop a kind of detectability and meet the requirements, favorable reproducibility is applicable to the residual immune colloid gold quick detection reagent plate of Methyltestosterone in the on-the-spot Rapid Screening aquatic products.
Another object of the present invention provides the preparation method of this agent plate.
A further object of the present invention provides the application of this agent plate.
Described agent plate is used chromatography type antibody mediated immunity competition principle, and by antigen and golden labeling antibody reaction solution, the Methyltestosterone in the specific detection aquatic products is residual.If sample solution contains residual, antibody response on Methyltestosterone elder generation and the colloid gold particle, therefore when colloid gold particle diffused to detection line with sample solution, the avtive spot of antibody can't be combined by the Methyltestosterone specific antigen on detection line because being occupied by the Methyltestosterone in the sample solution on the colloid gold particle; When the Methyltestosterone content in the sample surpassed the agent plate detection limit, the detection line colour developing on the agent plate was shallow even without colour developing than nature controlling line, is judged to be the positive.On the contrary, when Methyltestosterone content in the sample below the agent plate detection limit or during noresidue, the detection line colour developing on the agent plate is close with nature controlling line or partially dark, is judged to be feminine gender.
Described agent plate is comprised of sample pad, collaurum pad, nitrocellulose filter and the adsorptive pads closely pasted successively on up and down two plastic formworks, backing and the backings.Have in sample pad, collaurum pad, place that nitrocellulose filter is adjacent with adsorptive pads that 2mm ± 0.5mm's is overlapping, in order to guarantee chromatography effect carrying out smoothly from sample pad to the adsorptive pads position on the one hand, in order to make sample solution and sample be lined with sufficient reaction time on the other hand, buffer system on the sample pad can in and the potential of hydrogen of sample solution, guarantee that effective constituent in the sample solution and the golden labeling antibody on the collaurum pad react smoothly.
Each several part constituent and the function of described agent plate are as follows:
Plastic formwork plays fixedly test strips and referential function district (well, detection zone, Quality Control district).
Backing scribbles the toughness material that does not absorb water of adhesive sticker for one side, such as the PVC plate, play fixing other ingredients of test paper of supporting.
Sample pad is made by glass fibre, works to absorb sample solution and buffering sample solution pH value.
The collaurum pad is made by polyester film, is coated with the anti-Methyltestosterone monoclonal antibody of colloid gold label on it, is the place that effective constituent in the sample solution and golden labeling antibody react.
Nitrocellulose filter is coated with Methyltestosterone-bovine serum albumin(BSA) conjugate and sheep anti-mouse igg successively from sample pad to the adsorptive pads direction, respectively as detection line and nature controlling line, Main Function is with reaction result with macroscopic characterization out.
Adsorptive pads is filter paper, and its effect is the unnecessary solution absorption that comes up mobile as the suction part.
The preparation method of described agent plate comprises the steps:
(1) preparation of Methyltestosterone-carrier protein couplet thing: Methyltestosterone MT and carrier protein bovine serum albumin(BSA) BSA and ovalbumin OVA are carried out respectively coupling prepare envelope antigen MT-BSA and immunizing antigen MT-OVA;
(2) preparation of anti-Methyltestosterone monoclonal antibody: use immunizing antigen MT-OVA immunity to make anti-Methyltestosterone monoclonal antibody according to conventional method;
(3) use sodium citrate to make mean size and be the colloidal gold solution of 40nm;
(4) preparation of the anti-Methyltestosterone monoclonal antibody of colloid gold label: get the colloidal gold solution 100mL that has prepared, transfer pH to 8.0 with the 0.1mol/L solution of potassium carbonate; Add while stirring anti-Methyltestosterone monoclonal antibody 1.5mg, stir 20min, dropwise add again 2mL25mg/mL PEG 20000 (PEG20000), stir 15min; The centrifugal 15min of 20000r/min abandons supernatant, adds 10mL pH7.4PBS damping fluid (containing 0.4mg/mLPEG20000) and cleans 2 times; Precipitation is contained PBS damping fluid (pH7.4) dissolving of 2%BSA with 5mL, after filtering with 0.22 μ m sterilizing filter, 4 ℃ save backup;
(5) pre-service of collaurum pad and coated
Polyester film is cut into 85mm * 300mm, soaked 30 minutes with collaurum pad treating fluid, gently presses to remove excessive moisture with cylinder after taking out, place 15～37 ℃ drying box or drying room to dry more than 16 hours, it is for subsequent use that the placement environment temperature is that room temperature, humidity are lower than in the room of RH40%;
Collaurum pad treating fluid preparation method is: take by weighing 5g PVA, 7.1g Na
2HPO
4, 5g BSA, 1mL TritonX-100, add 1L H
2The O dissolving, regulating pH is 7.4;
Use some film instrument that the anti-Methyltestosterone monoclonal antibody of the colloid gold label for preparing is coated to pretreated collaurum pad, package amount is 2 μ L/cm;
(6) nitrocellulose filter is coated: with 0.01mol/L pH7.4, the PBS damping fluid that contains 4% trehalose and 2%NaCl MT-BSA and sheep anti-mouse igg are diluted to respectively 0.8 μ g/L and 1.2 μ g/L; Use that MT-BSA conjugate after some film instrument will dilute is coated to be arrived on 25mm * 300mm nitrocellulose filter as detection line, package amount is 1 μ L/cm; With the sheep anti-mouse igg after the dilution coated to the nitrocellulose filter as nature controlling line, package amount is 1 μ L/cm; Detection line is parallel with nature controlling line; 37 ℃ of dryings 8 hours;
(7) assembling of agent plate: sample pad and adsorptive pads are cut into 17mm * 300mm, the collaurum pad is cut into 8mm * 300mm; With well cutting sample pad, collaurum pad, nitrocellulose filter, adsorptive pads stick at successively on the backing, adjacent each several part has the overlapping of 2mm ± 0.5mm, forms the kilocalorie of 60mm * 300mm; Use cutting machine kilocalorie to be cut into the test strips of 3.84mm * 60mm, form agent plate in the plastic formwork of packing into.
The detection method of described agent plate comprises:
(1) sample pre-treatments: get a certain amount of tissue samples of chopping, use the homogenizer homogeneous; Take by weighing the 2.0g sample in the 5mL centrifuge tube; Add the 2mL absolute methanol, concuss 5min, the centrifugal 5min of 4000r/min under the room temperature condition; Draw supernatant 0.1mL in a new 1.5mL tubule, add the 0.4mL dilution, mixing, to be checked.
(2) horizontal positioned agent plate is drawn sample solution to be checked with dropper, and 3 (approximately 100 μ L) of vertical dropping begin timing behind the application of sample in well; The result should read at 3～5min, and the other times interpretation is invalid.
(3) result judges: during reading result, place the observer positive the agent plate level.
Negative (-): T line colour developing (detection line is near well one end) than C line (control line, i.e. nature controlling line) deeply or equally dark, Methyltestosterone concentration is lower than 10 μ g/kg or residual without Methyltestosterone in the expression sample.
Positive (+): the colour developing of T line is more shallow than C line, or the T line is without colour developing, and Methyltestosterone concentration is equal to or higher than 10 μ g/kg in the expression sample, and the colour developing of T line is more shallow than C line, and Methyltestosterone concentration is higher in the expression sample.
Invalid: the C line not occurring, may be that misoperation or agent plate lost efficacy.Should again read instructions, and retest with new agent plate.
The present invention has following beneficial effect:
(1) specificity is good, highly sensitive
The anti-Methyltestosterone monoclonal antibody of agent plate of the present invention is 100% to the cross reacting rate of Methyltestosterone, with the cross reacting rate of testosterone be 11.86%, with the cross reacting rate of 1-boldenone be 2.68%, with the cross reacting rate of testosterone propionate, 19-nortestosterone, diethylstilbestrol less than 1%.The detection limit of agent plate of the present invention can reach 10 μ g/kg, can satisfy the detection demand of aquatic product quality supervision fully.
(2) easy and simple to handle, quick
Most of raw material that agent plate of the present invention is reacted immunochromatography required has been incorporated in the agent plate, and antigen-antibody reaction is carried out on immobilon-p fast behind the sample, has greatly shortened the sample time, gets final product reading result in 3～5min behind the sample.The operation of this agent plate does not need any professional training, only needs to carry out sample pre-treatments by explanation with the naked eye interpretation testing result in several minutes behind the sample.
(3) dependence to experimental facilities is little
During Methyltestosterone, except needing to use in sample pretreatment process some light miniature instruments, other process does not all need to use instrument to agent plate of the present invention in detecting aquatic products.Drip after sample runs plate, the shade of judging detection line and nature controlling line on the nitrocellulose filter by naked eyes is to sentence read result recently, and this advantage makes it easy to field and execute-in-place.
(4) cost is low, and is profitable
Agent plate production technology of the present invention is simple, and the reagent that needs in the testing process is cheap, both can detect single sample, also is applicable to batch samples and detects, and production cost is low, greatly reduces testing cost.
Description of drawings
Fig. 1 is Methyltestosterone immune colloid gold quick detection reagent plate structure schematic diagram, and wherein 1 is sample pad, and 2 is the collaurum pad, and 3 is nitrocellulose filter, and 4 is detection line, and 5 is nature controlling line, and 6 is adsorptive pads, and 7 is adhesive sticker, and 8 is the PVC plate.
Fig. 2 is Methyltestosterone immune colloid gold quick detection reagent plate operation chart, and wherein S is well, and T is detection zone, and C is the Quality Control district.
Fig. 3,4,5 really judges schematic diagram for Methyltestosterone immune colloid gold quick detection reagent hardens, and wherein T is detection zone, and C is the Quality Control district; The negative result of Fig. 3, the positive result of Fig. 4, Fig. 5 is invalid.
Embodiment
The preparation method of Methyltestosterone immune colloid gold quick detection reagent plate
1, the preparation of Methyltestosterone-carrier protein couplet thing
The preparation of MT-BSA conjugate: take by weighing methyltestosterone 35mg, pyridine 1mL, ethyloic azanol half hydrochloride 24mg is put in the round-bottomed flask of 100mL, 50 ℃ of heating in water bath for reaction 30min, pyridine is removed in distillation.Residue 5mL acetic acid ethyl dissolution is washed three times, gets the upper strata oil sample, and the ethyl acetate that spends the night volatilizees.Get resultant product and dissolve with 250 μ L DMF, add 7.5 μ L triethylamines, 4 ℃ of lower reactions 1 hour add 12 μ L isobutyl chlorocarbonates subsequently, continue reaction 1 hour.Take by weighing 56mg BSA and be dissolved in the 500 μ L water, regulate pH to 8.5 with the NaOH of 1mol/L, add 500 μ L DMF, 4 ℃ of lower stirring reactions 1 hour, completely reacted product above then dropwise dripping, 4 ℃ of lower reactions 4 hours.Change water twice with the dialysis of PBS liquid, change water six times with the pure water dialysis again, water was changed once in 6 hours in the interval.
Adopt same method to prepare the MT-OVA conjugate.
2, the preparation of anti-Methyltestosterone monoclonal antibody
Get female Balb/c mouse in 6～8 ages in week, with MT-OVA conjugate and isopyknic Freund's complete adjuvant emulsification, press 80 μ g/ the subcutaneous multi-point injections in dosage back, every 2 all booster immunizations 1 time, use the Freunds incomplete adjuvant lumbar injection instead.The 3d reinforced immunological is 1 time before merging, and without adjuvant, dosage doubles.
The Sp2/0 multiple myeloma cells is mixed in 1: 10 ratio with immune spleen cell, merge under 50% polyglycol effect, the HAT nutrient culture media suspends, and divide and plant in 96 well culture plates, 37 ℃, 5%CO
2Cultivate in the incubator.The elisa plate of coated good antigen, wash 3 times with cleansing solution, then the culture supernatant that adds hybridoma is in 37 ℃ of incubation 1h, after cleansing solution is washed 3 times, add sheep anti-mouse igg-HRP, 37 ℃ of reaction 1h, wash 3 times after, add substrate OPD and develop the color, after the sulfuric acid cessation reaction, under the wavelength of 490nm, survey the OD value.Establish simultaneously the culture supernatant of Sp2/0 cell as negative control, OD
490Sample greater than 2 times of negative controls is positive, carries out cloning with limiting dilution assay.
Enlarge and cultivate building hybridoma after the strain, the whiteruss 0.5mL/ of lumbar injection sterilization only, 7～10d pneumoretroperitoneum injection hybridoma 5 * 10
5～10
6Individual/as only, to extract mouse ascites behind the 7d, the centrifuging and taking supernatant, mensuration is tired, and frozen.
3, the preparation of colloidal gold solution
In the 100mL deionized water, add the 1mL1% trisodium citrate, boil rear rapid adding 1mL1% gold chloride, continue to boil 10min, after the cooling, save backup under 4 ℃.The particle diameter of this colloid gold particle is about 40nm after measured.
4, the preparation of the anti-Methyltestosterone monoclonal antibody of colloid gold label
Get the colloidal gold solution 100mL that has prepared, transfer pH to 8.0 with the 0.1mol/L solution of potassium carbonate; Add while stirring anti-Methyltestosterone monoclonal antibody 1.5mg, stir 20min, dropwise add again 2mL25mg/mL PEG 20000 (PEG20000), stir 15min; The centrifugal 15min of 20000r/min abandons supernatant, adds 10mL pH7.4PBS damping fluid (containing 0.4mg/mLPEG20000) and cleans 2 times; Precipitation is contained PBS damping fluid (pH7.4) dissolving of 2%BSA with 5mL, after filtering with 0.22 μ m sterilizing filter, 4 ℃ save backup.
5, the pre-service of collaurum pad and coated
Polyester film is cut into 85mm * 300mm, soaked 30 minutes with collaurum pad treating fluid, gently presses to remove excessive moisture with cylinder after taking out, place 15～37 ℃ drying box or drying room to dry more than 16 hours, it is for subsequent use that the placement environment temperature is that room temperature, humidity are lower than in the room of RH40%;
Collaurum pad treating fluid preparation method is: take by weighing 5g PVA, 7.1g Na
2HPO
4, 5g BSA, 1mL TritonX-100, add 1L H
2The O dissolving, regulating pH is 7.4;
Use some film instrument that the anti-Methyltestosterone monoclonal antibody of the colloid gold label for preparing is coated to pretreated collaurum pad, package amount is 2 μ L/cm.
6, nitrocellulose filter is coated
With 0.01mol/L pH7.4, the PBS damping fluid that contains 4% trehalose and 2%NaCl MT-BSA and sheep anti-mouse igg are diluted to respectively 0.8 μ g/L and 1.2 μ g/L; Use that MT-BSA conjugate after some film instrument will dilute is coated to be arrived on 25mm * 300mm nitrocellulose filter as detection line, package amount is 1 μ L/cm; With the sheep anti-mouse igg after the dilution coated to the nitrocellulose filter as nature controlling line, package amount is 1 μ L/cm; Detection line is parallel with nature controlling line; 37 ℃ of dryings 8 hours.
7, the assembling of Methyltestosterone immune colloid gold quick detection reagent plate
Sample pad and adsorptive pads are cut into 17mm * 300mm, the collaurum pad is cut into 8mm * 300mm; With well cutting sample pad, collaurum pad, nitrocellulose filter, adsorptive pads stick at successively on the backing, adjacent each several part has the overlapping of 2mm ± 0.5mm, forms the kilocalorie of 60mm * 300mm; Use cutting machine kilocalorie to be cut into the test strips of 3.84mm * 60mm, form agent plate in the plastic formwork of packing into.
The performance evaluation of Methyltestosterone immune colloid gold quick detection reagent plate
1, the detection limit of Methyltestosterone immune colloid gold quick detection reagent plate test
Get the negative sample homogenate and subscript to MT content and be respectively 0,5,10,15,20,40 μ g/kg, carry out sample process and detection by above-mentioned sample treatment, each concentration is established 6 repetitions, drips plate and detects and the observation experiment result.Test findings shows: when the MT amount is 0,5 μ g/kg, the colour developing of T line be deeper than the colour developing of C line or and the colour developing of C line the same dark, when MT content is 10 μ g/kg and when above, the colour developing of T line is shallower than the C line and develops the color.Be limited to 10 μ g/kg therefore judge detecting of this agent plate.
2, the specific test of Methyltestosterone immune colloid gold quick detection reagent plate
Get the standard solution that MT, testosterone, testosterone propionate, 19-nortestosterone, 1-boldenone, diethylstilbestrol standard items make series concentration, detect with methyltestosterone immune colloid gold quick detection reagent plate, calculate cross reacting rate.
Test findings shows that this agent plate and MT cross reacting rate are 100%, with the cross reacting rate of testosterone be 11.86%, with the cross reacting rate of 1-boldenone be 2.68%, with the cross reacting rate of testosterone propionate, 19-nortestosterone, diethylstilbestrol less than 1%.
3, Methyltestosterone immune colloid gold quick detection reagent stability of plates test
Agent plate is placed in the environment that is positioned over respectively 37 ℃, 25 ℃, 4 ℃, and humidity keeps 40～60%, uses PBST solution to drip the plate reading after taking out at set intervals, and the colour developing of observing agent plate changes.
Test findings show agent plate 25 ℃ of lower holding times than 37 ℃ and 4 ℃ long, in 1 year the agent plate colour developing comparatively stable, meet testing requirement.
Claims (2)
1. the colloidal gold reagent plate of Methyltestosterone in the fast detecting aquatic products, it is characterized in that by two plastic formworks up and down, the sample pad of closely pasting successively on backing and the backing, the collaurum pad, nitrocellulose filter and adsorptive pads form, described backing scribbles the PVC plate of adhesive sticker for one side, described sample pad is made by glass fibre, described collaurum pad is made by polyester film, be coated with the anti-Methyltestosterone monoclonal antibody of colloid gold label on it, described nitrocellulose filter is coated with Methyltestosterone-bovine serum albumin(BSA) conjugate and sheep anti-mouse igg successively from sample pad to the adsorptive pads direction, as detection line and nature controlling line, described adsorptive pads is filter paper respectively.
2. the preparation method of the described agent plate of claim 1 is characterized in that, comprises the steps:
(1) preparation of Methyltestosterone-carrier protein couplet thing: Methyltestosterone MT and carrier protein bovine serum albumin(BSA) BSA and ovalbumin OVA are carried out respectively coupling prepare envelope antigen MT-BSA and immunizing antigen MT-OVA;
(2) preparation of anti-Methyltestosterone monoclonal antibody: use immunizing antigen MT-OVA immunity to make anti-Methyltestosterone monoclonal antibody according to conventional method;
(3) use sodium citrate to make mean size and be the colloidal gold solution of 40nm;
(4) preparation of the anti-Methyltestosterone monoclonal antibody of colloid gold label: get the colloidal gold solution 100mL that has prepared, transfer pH to 8.0 with the 0.1mol/L solution of potassium carbonate; Add while stirring anti-Methyltestosterone monoclonal antibody 1.5mg, stir 20min, dropwise add again 2mL25mg/mL PEG 20000 (PEG20000), stir 15min; The centrifugal 15min of 20000r/min abandons supernatant, adds 10mL pH7.4PBS damping fluid (containing 0.4mg/mL PEG20000) and cleans 2 times; Precipitation is contained PBS damping fluid (pH7.4) dissolving of 2%BSA with 5mL, after filtering with 0.22 μ m sterilizing filter, 4 ℃ save backup;
(5) pre-service of collaurum pad and coated
Polyester film is cut into 85mm * 300mm, soaked 30 minutes with collaurum pad treating fluid, gently presses to remove excessive moisture with cylinder after taking out, place 15～37 ℃ drying box or drying room to dry more than 16 hours, it is for subsequent use that the placement environment temperature is that room temperature, humidity are lower than in the room of RH40%;
Collaurum pad treating fluid preparation method is: take by weighing 5g PVA, 7.1g Na
2HPO
4, 5g BSA, 1mL TritonX-100, add 1L H
2The O dissolving, regulating pH is 7.4;
Use some film instrument that the anti-Methyltestosterone monoclonal antibody of the colloid gold label for preparing is coated to pretreated collaurum pad, package amount is 2 μ L/cm;
(6) nitrocellulose filter is coated: with 0.01mol/L pH7.4, the PBS damping fluid that contains 4% trehalose and 2%NaCl MT-BSA and sheep anti-mouse igg are diluted to respectively 0.8 μ g/L and 1.2 μ g/L; Use that MT-BSA conjugate after some film instrument will dilute is coated to be arrived on 25mm * 300mm nitrocellulose filter as detection line, package amount is 1 μ L/cm; With the sheep anti-mouse igg after the dilution coated to the nitrocellulose filter as nature controlling line, package amount is 1 μ L/cm; Detection line is parallel with nature controlling line; 37 ℃ of dryings 8 hours;
(7) assembling of agent plate: sample pad and adsorptive pads are cut into 17mm * 300mm, the collaurum pad is cut into 8mm * 300mm; With well cutting sample pad, collaurum pad, nitrocellulose filter, adsorptive pads stick at successively on the backing, adjacent each several part has the overlapping of 2mm ± 0.5mm, forms the kilocalorie of 60mm * 300mm; Use cutting machine kilocalorie to be cut into the test strips of 3.84mm * 60mm, form agent plate in the plastic formwork of packing into.
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Cited By (3)
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| CN103698524A (en) * | 2013-12-19 | 2014-04-02 | 杭州南开日新生物技术有限公司 | Immunity colloid gold reagent plate for quickly detecting sodium pentachlorophenate and preparation method for immunity colloid gold reagent plate |
| CN104558171A (en) * | 2014-12-26 | 2015-04-29 | 华中农业大学 | Monoclonal antibody, enzyme linked immunosorbent assay and kit for detecting methyltestosterone |
| CN109633182A (en) * | 2018-12-27 | 2019-04-16 | 深圳市宝安康生物技术有限公司 | Colloidal-gold detecting-card and preparation method thereof and hormone residues detection method |
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