CN106771215A - A kind of penicillin detection method and detection card - Google Patents
A kind of penicillin detection method and detection card Download PDFInfo
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- CN106771215A CN106771215A CN201611079065.5A CN201611079065A CN106771215A CN 106771215 A CN106771215 A CN 106771215A CN 201611079065 A CN201611079065 A CN 201611079065A CN 106771215 A CN106771215 A CN 106771215A
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- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
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- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/94—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving narcotics or drugs or pharmaceuticals, neurotransmitters or associated receptors
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- G—PHYSICS
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- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
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- G01N33/558—Immunoassay; Biospecific binding assay; Materials therefor using diffusion or migration of antigen or antibody
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Abstract
The invention discloses a kind of penicillin detection method and detection card, including reagent and instrument, the reagent includes penicillin sodium salt, PB buffer solutions, bovine serum albumin(BSA), glutaraldehyde solution, deionized water, trisodium citrate, gold chloride, solution of potassium carbonate, resistance to penicillin monoclonal antibody, PEG 20000, PH7.4PBS buffer solutions, PBS and sheep anti-mouse igg, and the instrument includes filter membrane, alcolhol burner, high speed freezing centrifuge, sterilizing filter, reserve tank and drying baker.The penicillin detection method and detection card, antigen-antibody reaction is quickly carried out on detection card after drop sample, substantially reduces the sample time, and sample, without specially treated, drop sample can be obtained a result in 36 minutes, and detection process is aided in without specific apparatus, ordinary person is operable, professional training is not needed, is easily used, construction is simple, low production cost, Detection accuracy is high, such that it is able to greatly improve the verification and measurement ratio of penicillin, and saves testing cost.
Description
Technical field
The present invention relates to penicillin technical field, specially a kind of penicillin detection method and detection block.
Background technology
Penicillin is otherwise known as benzyl penicillin, peillin G, penicillin, penicillin, Benzylpenicillin sodium salt, parasiticin
Sodium, benzylpenicillin potassium, potassium phenylacetamidopenicillanate.Penicillin is one kind of antibiotic, refer in molecule containing penam, bacterium can be destroyed
Cell membrane and from the breeding period of bacterial cell bactericidal action a class antibiotic, be the antibiotic by being extracted in Penicillium notatum.
Penicillin belongs to beta-lactam antibiotic, beta-lactam antibiotic include penicillin, cynnematin, Carbapenems,
Monocyclic class, cephamycin-type etc..Penicillin is the antimicrobial being in daily use.But skin test must be done before every time, with hypo-allergenic.
Most of penicillin detections are detected using high performance liquid chromatography at present, but these methods are excessively numerous
It is trivial, take it is more long, testing cost is higher, it is impossible to meet the requirement of quick detection, for this provide a kind of penicillin detection method and
Detection card.
The content of the invention
（One）The technical problem of solution
In view of the shortcomings of the prior art, the invention provides a kind of penicillin detection method and detection card, detection is solved time-consuming
Long and testing cost problem high.
（Two）Technical scheme
To achieve the above object, the present invention provides following technical scheme：A kind of penicillin detection card, including reagent and instrument, inspection
Blocking is surveyed to make to comprise the following steps：
S1, penicillin and the preparation of carrier protein couplet thing, take 40-60mg penicillin sodium salts and are dissolved in 1.5-2.5ml water, use 8-
12ml PB buffer solutions（0.1mol/L PH 6.0）Dissolving 40-60mg bovine serum albumin(BSA)s, are added in above-mentioned solution, then slowly
The glutaraldehyde solutions of 0.8-1.2ml 10% are added, mixed solution are stirred at room temperature 1.5-2.5h, dialysed 4-6 days with distilled water,
Filtered with filter membrane, collected；
S2, the preparation of colloidal gold solution takes the addition trisodium citrates of 1-2ml 1% in 100-200ml deionized waters, adds after boiling
Enter the gold chlorides of 1-2ml 1%, continue to boil 8-12min, after cooling, saved backup at 4 DEG C；
S3, the preparation of colloid gold label resistance to penicillin monoclonal antibody takes the 50-150ml colloidal gold solutions for having prepared completion, uses
Be adjusted to PH to 8.0 by 0.1mol/L solution of potassium carbonate, stirs above-mentioned solution while adding 1.0-2.0mg resistance to penicillin lists
It is anti-, 20-30min is stirred, then 1-3ml 25mol/L PEG 20000s are gradually dropped, and 12-18min is stirred, it is then above-mentioned molten
The liquid high speed freezing centrifuge of 20000rpm is centrifuged 15min, and reject supernatant liquor adds 8-12ml PH 7.4PBS buffer solutions
Cleaning twice, is dissolved with PBSs of the 4-6ml containing 2%BSA, and after being filtered with 0.22 μm of sterilizing filter, colloid gold label resists
Penicillin monoclonal antibody is saved backup at a temperature of 4 DEG C；
S4, the preparation of penicillin detection card, nitric acid is sprayed on by the penicillin carrier protein couplet thing and sheep anti-mouse igg of debita spissitudo
On cellulose membrane, respectively as detection line and control line, after the completion of, 7-9h is dried in 37 DEG C of drying baker, by debita spissitudo
Colloid gold label resistance to penicillin monoclonal antibody be sprayed on gold conjugation pad, then dry 7- in 37 DEG C of drying baker again
9h, nitrocellulose filter, gold conjugation pad, detection line, control line and adsorptive pads are stained with a base plate successively.
Preferably, the reagent includes penicillin sodium salt, PB buffer solutions, bovine serum albumin(BSA), glutaraldehyde solution, deionization
Water, trisodium citrate, gold chloride, solution of potassium carbonate, resistance to penicillin monoclonal antibody, PEG 20000, PH7.4PBS buffer solutions, PBS
Buffer solution and sheep anti-mouse igg.
Preferably, the instrument includes filter membrane, alcolhol burner, high speed freezing centrifuge, sterilizing filter, reserve tank and drying
Case.
A kind of penicillin detection method, is added dropwise on gold conjugation pad with sample solution to be tested, and sample solution is because of nitre
The capillarity of acid cellulose membrane carrier spreads to the other end, and during diffusion, antigen and antibody can occur accordingly
Antigen-antibody reaction, and it is real out by the color of immune colloid gold, if containing penicillin in sample solution, penicillin elder generation
With the antibody response on colloid gold particle, therefore when colloid gold particle diffuses to detection line with sample solution, colloid gold particle
Penicillin in the avtive spot of upper antibody sample solution is occupied and cannot work as sample with penicillin antigen binding in detection line
In Penicillin Content when exceeding the detection of detection card to greatest extent, the detection line colour developing on detection card is shallow compared with control line or even can show
Show colourless, be judged to the positive, when Penicillin Content is relatively low in sample, the upper detection line colour developing of detection card is close with control line even
Show Color is deeper, is judged to feminine gender.
Preferably, take in 1-4ml original milk addition 10ml centrifuge tubes, add 60 μ L 3mol/L HCI solution, vibrate 30-
40 seconds, 2-6ml ethyl acetate is added, acutely vibration 1-2 minutes, stratification goes 2-4ml upper solutions to another centrifuge tube
In, add 250 μ L PB buffer solutions（0.1mol/L PH 7.4）, vibrate 1-2 minutes, stratification, draw 80-150 μ L lower floors
Solution, takes out detection card, and solution to be checked is drawn with drip irrigation, and 2-4 drops are instilled on nitrocellulose filter, and timing is started after sample-adding,
Result should read after 3-6 minutes, and other times judge invalid.
（Three）Beneficial effect
The invention provides a kind of penicillin detection method and detection card, possesses following beneficial effect：
（1）The penicillin detection method and detection card, are incorporated on detection card by by required most of raw material, resist after drop sample
Antigen-antibody reaction is quickly carried out on detection card, substantially reduces sample time, and sample without specially treated, 3-6 points of drop sample
Can be obtained a result in clock, detection process is aided in without specific apparatus, ordinary person is operable, it is not necessary to professional training, pole
Easily use.
（2）The penicillin detection method and detection card, construction are simple, and low production cost, Detection accuracy is high, so that can
Greatly to improve the verification and measurement ratio of penicillin, and save testing cost.
Specific embodiment
Based on the embodiment in the present invention, those of ordinary skill in the art are obtained under the premise of creative work is not made
The every other embodiment for obtaining, belongs to the scope of protection of the invention.
The present invention provides a kind of technical scheme：A kind of penicillin detection card, including reagent and instrument, reagent include penicillin
Sodium salt, PB buffer solutions, bovine serum albumin(BSA), glutaraldehyde solution, deionized water, trisodium citrate, gold chloride, solution of potassium carbonate,
Resistance to penicillin monoclonal antibody, PEG 20000, PH7.4PBS buffer solutions, PBS and sheep anti-mouse igg, instrument include filter membrane,
Alcolhol burner, high speed freezing centrifuge, sterilizing filter, reserve tank and drying baker, detection blocking are made to comprise the following steps：
S1, penicillin and the preparation of carrier protein couplet thing, take 40-60mg penicillin sodium salts and are dissolved in 1.5-2.5ml water, use 8-
12ml PB buffer solutions（0.1mol/L PH 6.0）Dissolving 40-60mg bovine serum albumin(BSA)s, are added in above-mentioned solution, then slowly
The glutaraldehyde solutions of 0.8-1.2ml 10% are added, mixed solution are stirred at room temperature 1.5-2.5h, dialysed 4-6 days with distilled water,
Filtered with filter membrane, collected；
S2, the preparation of colloidal gold solution takes the addition trisodium citrates of 1-2ml 1% in 100-200ml deionized waters, adds after boiling
Enter the gold chlorides of 1-2ml 1%, continue to boil 8-12min, after cooling, saved backup at 4 DEG C；
S3, the preparation of colloid gold label resistance to penicillin monoclonal antibody takes the 50-150ml colloidal gold solutions for having prepared completion, uses
Be adjusted to PH to 8.0 by 0.1mol/L solution of potassium carbonate, stirs above-mentioned solution while adding 1.0-2.0mg resistance to penicillin lists
It is anti-, 20-30min is stirred, then 1-3ml 25mol/L PEG 20000s are gradually dropped, and 12-18min is stirred, it is then above-mentioned molten
The liquid high speed freezing centrifuge of 20000rpm is centrifuged 15min, and reject supernatant liquor adds 8-12ml PH 7.4PBS buffer solutions
Cleaning twice, is dissolved with PBSs of the 4-6ml containing 2%BSA, and after being filtered with 0.22 μm of sterilizing filter, colloid gold label resists
Penicillin monoclonal antibody is saved backup at a temperature of 4 DEG C；
S4, the preparation of penicillin detection card, nitric acid is sprayed on by the penicillin carrier protein couplet thing and sheep anti-mouse igg of debita spissitudo
On cellulose membrane, respectively as detection line and control line, after the completion of, 7-9h is dried in 37 DEG C of drying baker, by debita spissitudo
Colloid gold label resistance to penicillin monoclonal antibody be sprayed on gold conjugation pad, then dry 7- in 37 DEG C of drying baker again
9h, nitrocellulose filter, gold conjugation pad, detection line, control line and adsorptive pads are stained with a base plate successively.
A kind of penicillin detection method, detection method comprises the following steps：Take 1-4ml original milk add 10ml centrifuge tubes in,
60 μ L 3mol/L HCI solution are added, is vibrated 30-40 seconds, add 2-6ml ethyl acetate, acutely vibration 1-2 minutes, stood
Layering, goes 2-4ml upper solutions in another centrifuge tube, adds 250 μ L PB buffer solutions（0.1mol/L PH 7.4）, vibrate 1-
2 minutes, stratification drew 80-150 μ L lower floors solution, takes out detection card, solution to be checked is drawn with drip irrigation, in cellulose nitrate
2-4 drops are instilled on plain film, timing is started after sample-adding, as a result should read after 3-6 minutes, other times judge invalid, with to be checked
Test sample solution to be added dropwise on gold conjugation pad, sample solution is because the capillarity of nitrocellulose membrane carrier is to the other end
Diffusion, during diffusion, antigen and antibody can occur corresponding antigen-antibody reaction, and the color for passing through immune colloid gold
Out, if containing penicillin in sample solution, antibody response of the penicillin first and on colloid gold particle therefore works as colloid to reality
When gold grain diffuses to detection line with sample solution, the penicillin on colloid gold particle in the avtive spot of antibody sample solution
Occupy and cannot be detected to greatest extent when the Penicillin Content in sample exceedes detection card with penicillin antigen binding in detection line
When, the detection line colour developing on detection card is shallow compared with control line or even can show colourless, is judged to the positive, when Penicillin Content in sample
When relatively low, detection card upper detection line colour developing or even Show Color close with control line is deeper, is judged to feminine gender.
Embodiment 1
A kind of penicillin detection method, detection method comprises the following steps：
S1, penicillin and the preparation of carrier protein couplet thing, take 40mg penicillin sodium salts and are dissolved in 1.5ml water, slow with 8ml PB
Fliud flushing（0.1mol/L PH 6.0）Dissolving 40mg bovine serum albumin(BSA)s, are added in above-mentioned solution, are slow added into 0.8ml
10% glutaraldehyde solution, 1.5h is stirred at room temperature by mixed solution, is dialysed 4 days with distilled water, is filtered with filter membrane, is collected；
S2, the preparation of colloidal gold solution takes the addition trisodium citrates of 1ml 1% in 100ml deionized waters, and 1ml is added after boiling
1% gold chloride, continues to boil 8min, after cooling, is saved backup at 4 DEG C；
S3, the preparation of colloid gold label resistance to penicillin monoclonal antibody takes the 50ml colloidal gold solutions for having prepared completion, uses
Be adjusted to PH to 8.0 by 0.1mol/L solution of potassium carbonate, stirs above-mentioned solution while adding 1.0mg resistance to penicillin monoclonal antibodies, stirs
20min is mixed, then is gradually dropped 1ml 25mol/L PEG 20000s, stir 12min, then above-mentioned solution is with 20000rpm's
High speed freezing centrifuge is centrifuged 15min, and reject supernatant liquor adds 8ml PH 7.4PBS buffer solution for cleaning twice, contained with 4ml
The PBS dissolving of 2%BSA, after being filtered with 0.22 μm of sterilizing filter, colloid gold label resistance to penicillin monoclonal antibody is 4
Saved backup at a temperature of DEG C；
S4, the preparation of penicillin detection card, nitric acid is sprayed on by the penicillin carrier protein couplet thing and sheep anti-mouse igg of debita spissitudo
On cellulose membrane, respectively as detection line and control line, after the completion of, 7h is dried in 37 DEG C of drying baker, by debita spissitudo
Colloid gold label resistance to penicillin monoclonal antibody is sprayed on gold conjugation pad, then dries 7h in 37 DEG C of drying baker again,
Nitrocellulose filter, gold conjugation pad, detection line, control line and adsorptive pads are stained with successively on one base plate；
Take in 1ml original milk addition 10ml centrifuge tubes, add 60 μ L 3mol/L HCI solution, vibrate 30 seconds, add 2ml acetic acid
Ethyl ester, acutely vibration 1 minute, stratification, go 2ml upper solutions in another centrifuge tube, add 250 μ L PB buffer solutions
（0.1mol/L PH 7.4）, vibrate 1 minute, stratification, 80 μ L lower floors solution are drawn, detection card is taken out, drawn with drip irrigation and treated
Inspection solution, instills 2 drops on nitrocellulose filter, and timing is started after sample-adding, as a result should read after 3 minutes, and other times are sentenced
It is disconnected invalid.
Embodiment 2
A kind of penicillin detection method, detection method comprises the following steps：
S1, penicillin and the preparation of carrier protein couplet thing, take 50mg penicillin sodium salts and are dissolved in 2ml water, are buffered with 10ml PB
Liquid（0.1mol/L PH 6.0）Dissolving 50mg bovine serum albumin(BSA)s, are added in above-mentioned solution, are slow added into 1.0ml 10%
Glutaraldehyde solution, 2h is stirred at room temperature by mixed solution, is dialysed 5 days with distilled water, is filtered with filter membrane, is collected；
S2, the preparation of colloidal gold solution takes the addition trisodium citrates of 1.5ml 1% in 150ml deionized waters, is added after boiling
The gold chlorides of 1.5ml 1%, continue to boil 10min, after cooling, are saved backup at 4 DEG C；
S3, the preparation of colloid gold label resistance to penicillin monoclonal antibody takes the 100ml colloidal gold solutions for having prepared completion, uses
Be adjusted to PH to 8.0 by 0.1mol/L solution of potassium carbonate, stirs above-mentioned solution while adding 1.5mg resistance to penicillin monoclonal antibodies, stirs
25min is mixed, then is gradually dropped 2ml 25mol/L PEG 20000s, stir 15min, then above-mentioned solution is with 20000rpm's
High speed freezing centrifuge is centrifuged 15min, and reject supernatant liquor adds 10ml PH 7.4PBS buffer solution for cleaning twice, contained with 5ml
The PBS dissolving of 2%BSA, after being filtered with 0.22 μm of sterilizing filter, colloid gold label resistance to penicillin monoclonal antibody is 4
Saved backup at a temperature of DEG C；
S4, the preparation of penicillin detection card, nitric acid is sprayed on by the penicillin carrier protein couplet thing and sheep anti-mouse igg of debita spissitudo
On cellulose membrane, respectively as detection line and control line, after the completion of, 8h is dried in 37 DEG C of drying baker, by debita spissitudo
Colloid gold label resistance to penicillin monoclonal antibody is sprayed on gold conjugation pad, then dries 8h in 37 DEG C of drying baker again,
Nitrocellulose filter, gold conjugation pad, detection line, control line and adsorptive pads are stained with successively on one base plate；
Take in 2.5ml original milk addition 10ml centrifuge tubes, add 60 μ L 3mol/L HCI solution, vibrate 35 seconds, add 4ml second
Acetoacetic ester, acutely vibration 1.5 minutes, stratification go 3ml upper solutions in another centrifuge tube, adding 250 μ L PB bufferings
Liquid（0.1mol/L PH 7.4）, vibrate 1.5 minutes, stratification, 120 μ L lower floors solution are drawn, detection card is taken out, use drip irrigation
Solution to be checked is drawn, 3 is instilled on nitrocellulose filter and is dripped, timing is started after sample-adding, as a result should read after 4 minutes, other
Time judges invalid.
Embodiment 3
A kind of penicillin detection method, detection method comprises the following steps：
S1, penicillin and the preparation of carrier protein couplet thing, take 60mg penicillin sodium salts and are dissolved in 2.5ml water, slow with 12ml PB
Fliud flushing（0.1mol/L PH 6.0）Dissolving 60mg bovine serum albumin(BSA)s, are added in above-mentioned solution, are slow added into 1.2ml
10% glutaraldehyde solution, 2.5h is stirred at room temperature by mixed solution, is dialysed 6 days with distilled water, is filtered with filter membrane, is collected；
S2, the preparation of colloidal gold solution takes the addition trisodium citrates of 2ml 1% in 200ml deionized waters, and 2ml is added after boiling
1% gold chloride, continues to boil 12min, after cooling, is saved backup at 4 DEG C；
S3, the preparation of colloid gold label resistance to penicillin monoclonal antibody takes the 150ml colloidal gold solutions for having prepared completion, uses
Be adjusted to PH to 8.0 by 0.1mol/L solution of potassium carbonate, stirs above-mentioned solution while adding 2.0mg resistance to penicillin monoclonal antibodies, stirs
30min is mixed, then is gradually dropped 3ml 25mol/L PEG 20000s, stir 18min, then above-mentioned solution is with 20000rpm's
High speed freezing centrifuge is centrifuged 15min, and reject supernatant liquor adds 12ml PH 7.4PBS buffer solution for cleaning twice, contained with 6ml
The PBS dissolving of 2%BSA, after being filtered with 0.22 μm of sterilizing filter, colloid gold label resistance to penicillin monoclonal antibody is 4
Saved backup at a temperature of DEG C；
S4, the preparation of penicillin detection card, nitric acid is sprayed on by the penicillin carrier protein couplet thing and sheep anti-mouse igg of debita spissitudo
On cellulose membrane, respectively as detection line and control line, after the completion of, 9h is dried in 37 DEG C of drying baker, by debita spissitudo
Colloid gold label resistance to penicillin monoclonal antibody is sprayed on gold conjugation pad, then dries 9h in 37 DEG C of drying baker again,
Nitrocellulose filter, gold conjugation pad, detection line, control line and adsorptive pads are stained with successively on one base plate；
Take in 4ml original milk addition 10ml centrifuge tubes, add 60 μ L 3mol/L HCI solution, vibrate 40 seconds, add 6ml acetic acid
Ethyl ester, acutely vibration 2 minutes, stratification, go 4ml upper solutions in another centrifuge tube, add 250 μ L PB buffer solutions
（0.1mol/L PH 7.4）, vibrate 2 minutes, stratification, 150 μ L lower floors solution are drawn, detection card is taken out, drawn with drip irrigation
Solution to be checked, instills 4 drops on nitrocellulose filter, and timing is started after sample-adding, as a result should be read after 6 minutes, other times
Judge invalid.
Wherein：Refrigerated centrifuge is exactly using centrifugal force so that needing separate different material to obtain the machine for accelerating separate
Device, its model GL-20C.
In sum, the penicillin detection method and detection card, when the Penicillin Content in sample exceedes detection card detection
When to greatest extent, the detection line colour developing on detection card is shallow compared with control line or even can show colourless, is judged to the positive, when blue or green in sample
When mycin content is relatively low, detection card upper detection line colour developing or even Show Color close with control line is deeper, is judged to feminine gender.
It should be noted that herein, such as first and second or the like relational terms are used merely to a reality
Body or operation make a distinction with another entity or operation, and not necessarily require or imply these entities or deposited between operating
In any this actual relation or order.And, term " including ", "comprising" or its any other variant be intended to
Nonexcludability is included, so that process, method, article or equipment including a series of key elements not only will including those
Element, but also other key elements including being not expressly set out, or also include being this process, method, article or equipment
Intrinsic key element.In the absence of more restrictions.By sentence " including one ... the key element for limiting, it is not excluded that
Also there is other identical element in the process including the key element, method, article or equipment ".
Although an embodiment of the present invention has been shown and described, for the ordinary skill in the art, can be with
Understanding can carry out various changes, modification, replacement to these embodiments without departing from the principles and spirit of the present invention
And modification, the scope of the present invention be defined by the appended.
Claims (5)
1. a kind of penicillin detection blocks, including reagent and instrument, it is characterised in that：Detection blocking is made to comprise the following steps：
S1, penicillin and the preparation of carrier protein couplet thing, take 40-60mg penicillin sodium salts and are dissolved in 1.5-2.5ml water, use 8-
12ml PB buffer solutions（0.1mol/L PH 6.0）Dissolving 40-60mg bovine serum albumin(BSA)s, are added in above-mentioned solution, then slowly
The glutaraldehyde solutions of 0.8-1.2ml 10% are added, mixed solution are stirred at room temperature 1.5-2.5h, dialysed 4-6 days with distilled water,
Filtered with filter membrane, collected；
S2, the preparation of colloidal gold solution takes the addition trisodium citrates of 1-2ml 1% in 100-200ml deionized waters, adds after boiling
Enter the gold chlorides of 1-2ml 1%, continue to boil 8-12min, after cooling, saved backup at 4 DEG C；
S3, the preparation of colloid gold label resistance to penicillin monoclonal antibody takes the 50-150ml colloidal gold solutions for having prepared completion, uses
Be adjusted to PH to 8.0 by 0.1mol/L solution of potassium carbonate, stirs above-mentioned solution while adding 1.0-2.0mg resistance to penicillin lists
It is anti-, 20-30min is stirred, then 1-3ml 25mol/L PEG 20000s are gradually dropped, and 12-18min is stirred, it is then above-mentioned molten
The liquid high speed freezing centrifuge of 20000rpm is centrifuged 15min, and reject supernatant liquor adds 8-12ml PH 7.4PBS buffer solutions
Cleaning twice, is dissolved with PBSs of the 4-6ml containing 2%BSA, and after being filtered with 0.22 μm of sterilizing filter, colloid gold label resists
Penicillin monoclonal antibody is saved backup at a temperature of 4 DEG C；
S4, the preparation of penicillin detection card, nitric acid is sprayed on by the penicillin carrier protein couplet thing and sheep anti-mouse igg of debita spissitudo
On cellulose membrane, respectively as detection line and control line, after the completion of, 7-9h is dried in 37 DEG C of drying baker, by debita spissitudo
Colloid gold label resistance to penicillin monoclonal antibody be sprayed on gold conjugation pad, then dry 7- in 37 DEG C of drying baker again
9h, nitrocellulose filter, gold conjugation pad, detection line, control line and adsorptive pads are stained with a base plate successively.
2. a kind of penicillin detection according to claim 1 blocks, it is characterised in that：The reagent include penicillin sodium salt,
PB buffer solutions, bovine serum albumin(BSA), glutaraldehyde solution, deionized water, trisodium citrate, gold chloride, solution of potassium carbonate, anti-mould
Plain monoclonal antibody, PEG 20000, PH7.4PBS buffer solutions, PBS and sheep anti-mouse igg.
3. a kind of penicillin detection according to claim 1 blocks, it is characterised in that：The instrument include filter membrane, alcolhol burner,
High speed freezing centrifuge, sterilizing filter, reserve tank and drying baker.
4. a kind of penicillin detection method, it is characterised in that：It is added dropwise on gold conjugation pad with sample solution to be tested, sample
Solution spreads because of the capillarity of nitrocellulose membrane carrier to the other end, and during diffusion, antigen and antibody can be sent out
The corresponding antigen-antibody reaction of life, and it is real out by the color of immune colloid gold, if containing penicillin in sample solution,
Antibody response of the penicillin first and on colloid gold particle, therefore when colloid gold particle diffuses to detection line with sample solution, glue
Penicillin on body gold grain in the avtive spot of antibody sample solution occupies and cannot be with penicillin antigen knot in detection line
Close, when the Penicillin Content in sample exceedes the detection of detection card to greatest extent, the detection line on detection card develops the color compared with control line
It is shallow or even can show colourless, it is judged to the positive, when Penicillin Content is relatively low in sample, the upper detection line colour developing of detection card and control
Line is close or even Show Color is deeper, is judged to feminine gender.
5. a kind of penicillin detection method according to claim 4, it is characterised in that：Take 1-4ml original milk add 10ml from
In heart pipe, 60 μ L 3mol/L HCI solution are added, vibrated 30-40 seconds, add 2-6ml ethyl acetate, acutely 1-2 points of vibration
Clock, stratification goes 2-4ml upper solutions in another centrifuge tube, adds 250 μ L PB buffer solutions（0.1mol/L PH
7.4）, vibrate 1-2 minutes, stratification, 80-150 μ L lower floors solution is drawn, detection card is taken out, solution to be checked is drawn with drip irrigation,
2-4 drops are instilled on nitrocellulose filter, timing is started after sample-adding, as a result should read after 3-6 minutes, other times judge nothing
Effect.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201611079065.5A CN106771215A (en) | 2016-11-30 | 2016-11-30 | A kind of penicillin detection method and detection card |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201611079065.5A CN106771215A (en) | 2016-11-30 | 2016-11-30 | A kind of penicillin detection method and detection card |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN106771215A true CN106771215A (en) | 2017-05-31 |
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| CN201611079065.5A Pending CN106771215A (en) | 2016-11-30 | 2016-11-30 | A kind of penicillin detection method and detection card |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111665355A (en) * | 2020-05-06 | 2020-09-15 | 量准（上海）医疗器械有限公司 | Kit based on nano plasma resonance molecules and testing method |
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| CN101710121A (en) * | 2009-11-06 | 2010-05-19 | 杭州南开日新生物技术有限公司 | Method for preparing immunological colloidal gold reagent plate used for detecting penicillin |
| CN201535777U (en) * | 2009-09-02 | 2010-07-28 | 杭州南开日新生物技术有限公司 | Immunocolloid gold reagent plate for detecting penicillin |
| CN102323415A (en) * | 2011-06-13 | 2012-01-18 | 清华大学深圳研究生院 | Immunochromatography test paper for detecting Ciprofloxacin and preparation method thereof |
| CN103105492A (en) * | 2013-01-17 | 2013-05-15 | 重庆市科学技术研究院 | Fluorescence immunochromatography flu virus detection test paper |
| CN203720183U (en) * | 2014-02-15 | 2014-07-16 | 北京勤邦生物技术有限公司 | Clebuterol hydrochloride fluorescent microsphere immunochromatography test strip |
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| WO2001025795A2 (en) * | 1999-10-04 | 2001-04-12 | Dsm N.V. | One step test to detect antimicrobial residues in eggs |
| CN201535777U (en) * | 2009-09-02 | 2010-07-28 | 杭州南开日新生物技术有限公司 | Immunocolloid gold reagent plate for detecting penicillin |
| CN101710121A (en) * | 2009-11-06 | 2010-05-19 | 杭州南开日新生物技术有限公司 | Method for preparing immunological colloidal gold reagent plate used for detecting penicillin |
| CN102323415A (en) * | 2011-06-13 | 2012-01-18 | 清华大学深圳研究生院 | Immunochromatography test paper for detecting Ciprofloxacin and preparation method thereof |
| CN103105492A (en) * | 2013-01-17 | 2013-05-15 | 重庆市科学技术研究院 | Fluorescence immunochromatography flu virus detection test paper |
| CN203720183U (en) * | 2014-02-15 | 2014-07-16 | 北京勤邦生物技术有限公司 | Clebuterol hydrochloride fluorescent microsphere immunochromatography test strip |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN111665355A (en) * | 2020-05-06 | 2020-09-15 | 量准（上海）医疗器械有限公司 | Kit based on nano plasma resonance molecules and testing method |
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