CN102539747A - Enzyme-linked immunoassay kit for detecting phenylethanolamine A by direct competition method - Google Patents
Enzyme-linked immunoassay kit for detecting phenylethanolamine A by direct competition method Download PDFInfo
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- CN102539747A CN102539747A CN2011104455178A CN201110445517A CN102539747A CN 102539747 A CN102539747 A CN 102539747A CN 2011104455178 A CN2011104455178 A CN 2011104455178A CN 201110445517 A CN201110445517 A CN 201110445517A CN 102539747 A CN102539747 A CN 102539747A
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Abstract
The invention discloses an enzyme-linked immunoassay kit for detecting phenylethanolamine A by a direct competition method. The enzyme-linked immunoassay kit for detecting phenylethanolamine A provided by the invention comprises an enzyme label plate coated by phenylethanolamine A specific antibodies, and a phenylethanolamine A horseradish peroxidase marker. The enzyme-linked immunoassay kit for detecting phenylethanolamine A of the invention is sensitive, rapid, accurate, and is mainly used for screening of large quantities of samples; main reagents in the kit are provided in the form of operating fluid, and the kit is convenient for using, has the characteristics of high specificity, high sensitivity, high precision, high accuracy, and the like, can rapidly detect residual phenylethanolamine A in feed and animal products.
Description
Technical field
The present invention relates to enzyme linked immunological and detection of veterinary drugs in food field.Particularly, the present invention relates to a kind of enzyme linked immunological kit that is used to detect phenolethanolamine A.
Technical background
Phenolethanolamine A is called Ke Lunba amine (clenrapmine CLRA) again, and formal name used at school is 2-4-(4-nitrobenzophenone) butyl-2-base amino-1-methoxybenzene ethanol, is a kind of chemical substance of synthetic.Ke Lunba amine is the isomers of Formoterol, is the accessory substance of the synthetic Ractopamine of Lilly Co., Eli., has with clenbuterol hydrochloride effect and the effect identical with Ractopamine, belongs to a kind of of β 2-activator.Classified as the material of forbidding use in feed and animal drinking-water by No. 1519 bulletin of the Ministry of Agriculture.Therefore must set up detection method, forbid the generation of this type of incident, ensure food safety.
Mostly existing phenolethanolamine A detection method is chromatography, but the sensitivity of these methods receive sample purification, step such as concentrate influence bigger; Moreover the complicated instrument of these methods needs, and process is loaded down with trivial details, the examination of incompatible on-the-spot great amount of samples.For the analyzing and testing of phenolethanolamine A residue screening method is provided based on the immunoreactive immunology detection technology of antigen-antibody.The key of this technical research is the design of hapten molecule, the preparation of synthetic and artificial holoantigen and antibody.
Because phenolethanolamine A is a micromolecular compound, itself does not have immunogenicity, must itself and macromolecular carrier albumen coupling be obtained having immunogenic artificial antigen.The ELISA method of phenolethanolamine A is not appeared in the newspapers as yet at present.
Summary of the invention
The object of the present invention is to provide the enzyme linked immunological kit of a kind of fast detecting phenolethanolamine A.
The invention provides the direct competition method enzyme linked immunological kit of a kind of fast detecting phenolethanolamine A, this kit comprises: the ELISA Plate, the phenolethanolamine A-horseradish peroxidase-labeled thing that are coated with phenolethanolamine A specific antibody.Wherein phenolethanolamine A specific antibody is the polyclonal antibody of the anti-phenolethanolamine A of rabbit, and available phenolethanolamine A and carrier protein couplet thing make through immune animal (for example rabbit, sheep, mouse etc.) as immunogene.Described carrier protein is the blue albumen (KLH) of bovine serum albumin(BSA), human serum albumins, ovalbumin or key hole copper.Described phenolethanolamine A-horseradish peroxidase-labeled thing adopts chemical method that phenolethanolamine A and horseradish peroxidase are obtained.
Be used to prepare the solid phase material of said ELISA Plate, include but not limited to, for example, polystyrene, tygon, polypropylene.The form of carrier is the micro-reaction plate shrinkage pool.
Be convenient on-the-spot the detection and the great amount of samples examination, said kit can further include phenolethanolamine A-horseradish peroxidase working fluid, phenolethanolamine A standard solution, developer, cleansing solution, stop buffer and concentrating sample dilution.
Described cleansing solution is a 0.05%-0.5% Tween-20 phosphate buffer.Described developer is formed (for example, volume ratio is 1: 1) by developer A liquid and developer B liquid, and developer A liquid is hydrogen peroxide or urea peroxide, and developer B liquid is o-phenylenediamine or tetramethyl benzidine.Described concentrating sample dilution is the phosphate buffer that contains 0.1% Tween-20.
On the other hand, the present invention also provides a kind of method of examining phenolethanolamine A content in the feed test sample article, comprises step:
(1) sample pre-treatments;
(2) detect with the arbitrary described kit of claim 1-7; In the ELISA Plate hole that is coated with phenolethanolamine A antibody, add standard items or sample solution; Add phenolethanolamine A-horseradish peroxidase working fluid again, hatch back washing bat and do, add developer; Hatch the back and stop, measure absorbance with ELIASA;
(3) analyzing and testing result.
The detection principle of kit of the present invention is:
Phenolethanolamine A antibody is adsorbed on the solid phase carrier; Add sample or phenolethanolamine A standard solution and add phenolethanolamine A-horseradish peroxidase working fluid; Phenolethanolamine A antibody in the testing sample on phenolethanolamine A and the phenolethanolamine A-horseradish peroxidase competition ELISA Plate, the colour developing back stops the absorbance of working sample; The amount of phenolethanolamine A is negative correlation in this value and the sample, relatively can draw phenolethanolamine A concentration range with typical curve.
Beneficial effect:
The kit that the present invention detects phenolethanolamine A mainly adopts the direct competitive enzymoimmunoassay, qualitatively or quantitatively determines phenolethanolamine A content in the sample; Pre-treatment requirement to sample is low, and sample pretreatment process is simple, simultaneously the fast detecting gross sample.
This kit of the present invention adopts the phenolethanolamine A polyclonal antibody of high specific; Main agents provides with the working fluid form; Can reduce the operation steps of kit; For the user saves time and reduces the error that causes because of operation steps is miscellaneous; That the present invention has is highly sensitive, high specificity, pinpoint accuracy, pin-point accuracy, low to the instrument and equipment requirement, the reagent holding time is long, automaticity is high, "dead" isotopic contamination etc. advantage, can in feed and animal derived product detect, play a significant role.
Description of drawings
Accompanying drawing suppresses curve for phenolethanolamine A.
Specific embodiments
It is in order further to understand the present invention better that following embodiment is provided, and never content of the present invention and protection domain is constituted any restriction.
The embodiment 1 immunogenic synthetic preparation that reaches immune serum
1.1 reagent and instrument
Phenolethanolamine A (DNA Sci-tech Co., Ltd. is so kind as to give), succinic anhydride (available from Beijing northern Bioisystech Co., Ltd that shines), pyridine (Pyridine; AR.WM=79.10, content＞99.5%, Tianjin section close europeanized reagent development centre); N, dinethylformamide (Dimethylformamide, DMF; N.J. ACROSORGANICS company produces, available from lark prestige chemical reagents corporation), EDC (available from lark prestige chemical reagents corporation); Bovine serum albumin(BSA) (BSA), ovalbumin (OVA) etc. (available from Beijing northern Bioisystech Co., Ltd that shines), other reagent be analyze pure.
Twin-beam UV, visible light spectrophotometer (TU-1909, the general all purpose instrument company limited of analysing in Beijing), chromatographic apparatus (3057 type potable recording appearance, river, Chongqing appearance four factories; SBS series numerical control drop recorder, constant flow pump, fraction collector automatically; Chromatographic column, Shanghai Hu Xi analytical instrument factory), magnetic stirring apparatus (east, Shanghai Rong Feng scientific instrument company limited); Desk centrifuge (Minispin maximum (top) speed 13400rpm maximum centrifugal force 12100rcf, 2ml * 12)
1.2 phenolethanolamine A artificial antigen is synthetic
Take by weighing phenolethanolamine A 3.4mg (0.01mmol) and be dissolved in the 0.4ml pyridine solution, add the reaction of succinic anhydride 4mg stirring at room and spend the night.
Dry up pyridine, with the phenolethanolamine A of 0.2mlDMF dissolving acid anhydrides activation, the phenolethanolamine A solution of activation dropwise joins in 0.1M pH 7.5 ice-cold carrier proteins (BSA, OVA or KLH) the PBS solution, adds EDC 20mg, and stirring reaction spends the night.
Conjugate is crossed the SephadexG-25M gel chromatography; With three times of bed volume balances of 0.01M pH 7.4PBS; Regulate flow velocity to 3ml/min, sample concentration joins chromatography purification conjugate in the good chromatographic column of balance to 5ml, and eluent is with pH 7.4 0.01M PBS.
1.3 phenolethanolamine A horseradish peroxidase-labeled thing is synthetic
Take by weighing phenolethanolamine A 3.4mg (0.01mmol) and be dissolved in the 0.4ml pyridine solution, add the reaction of succinic anhydride 4mg stirring at room and spend the night.
Dry up pyridine, with the phenolethanolamine A of 0.2mlDMF dissolving acid anhydrides activation, the phenolethanolamine A solution of activation dropwise joins in 0.1M pH 7.5 horseradish peroxidases (HRP) the PBS solution, adds EDC 20mg, and stirring reaction spends the night.
Conjugate is crossed the SephadexG-25M gel chromatography; With three times of bed volume balances of 0.01M pH 7.4PBS; Regulate flow velocity to 3ml/min, sample concentration joins chromatography purification conjugate in the good chromatographic column of balance to 5ml, and eluent is with pH 7.4 0.01M PBS.
1.4 the preparation of immunity and specific antibody
Above-mentioned phenolethanolamine A-BSA conjugate is diluted to the 1mg/ml solution for standby with physiological saline.
Choose 6 body weight 2～2.5Kg healthy male new zealand white rabbits.Conjugate and equivalent Freund's complete adjuvant are mixed into water in oil emulsion through syringe to the method for taking out, carry out first immunisation, take the subcutaneous multi-point injection in back by the amount of 1mg/Kg body weight.Whenever once, replace Freund's complete adjuvant with incomplete Freund, the same first immunisation of dosage and method at a distance from two all booster immunizations.From immunity beginning for the third time, back 10 days of each immunity, auricular vein is got blood 1ml, carries out antibody titer and detects; When antibody titer no longer raises, do not add adjuvant and carry out for the last time (the 7th time) immunity, leg muscle injection, rear neck artery bloodletting in 7 days; Room temperature is solidified behind the 2h 4 ℃ and is spent the night, and centrifugal 10 minutes of 8000r/min removes clot, and serum partly precipitates with 50% saturated ammonium sulfate solution; The centrifugal supernatant that goes, precipitate resuspended with phosphate buffer, again with twice of 33% saturated ammonium sulfate solution deposition; Sediment is used the SephadexG-25M chromatography with the least possible phosphate buffer dissolving after dialysis, promptly get polyclonal antibody.
The foundation of embodiment 2 immunologic detection methods
2.1 ELISA square formation method is confirmed optimum response concentration
The phenolethanolamine A antibody of 1000 μ g/ml, 100 μ g/ml, 10 μ g/ml, 5 μ g/ml, 1 μ g/ml, 0.25 μ g/ml series concentration is pressed every hole 100 μ l coated elisa plates; 4 ℃ encapsulate and spend the night; Wash 3 times, clap and do, seal down for 4 ℃ by every hole 200 μ l confining liquids and spend the night; Wash 3 times, clap and do.Adding is since the phenolethanolamine A horseradish peroxidase-labeled thing 50ul of 1: 100 doubling dilution, and room temperature effect 45 minutes is washed three times; Add 50 μ l substrate A liquid and 50 μ l substrate B liquid; Room temperature lucifuge effect 15min, 50 μ l stop buffer cessation reactions, ELIASA detects A value (450nm).Be provided with simultaneously two parallel, getting the encapsulate concentration of OD value when being 1.5 left and right sides is optium concentration.Test figure is listed in table 3.
Table 1 square formation method OD value
By confirming in the data of table 3 that it is 5 μ g/ml that optimum antibody encapsulates concentration.The enzyme-labelled antigen extension rate reaches optimum response concentration at 1: 200.
2.2 ELISA detects antibody I C50 value
The ELISA method of operating is with 2.1, and it is 5 μ g/ml that optimum antibody encapsulates concentration, and enzyme-labelled antigen extension rate 1: 200 is prepared the free phenolethanolamine A standard items of 0 μ g/L, 0.1 μ g/L, 0.3 μ g/L, 0.9 μ g/L, 2.7 μ g/L, 8.1 μ g/L series concentration.Every hole adds 50 μ l standard items, adds 50 μ l enzyme-labelled antigen dilutions subsequently, and room temperature reaction 45 minutes is washed plate 3 times, adds the developer incubation 15 minutes, cessation reaction, ELIASA reading.The data of testing result see the following form
Can know phenolethanolamine A antiserum IC by data in the table
50Value shows that the antiserum that the present invention prepares is to have specific preferably anti-phenolethanolamine A polyclonal antibody about 1ng/ml, can satisfy phenolethanolamine A and detect requirement.
Set up the enzyme linked immunological kit that detects phenolethanolamine A, make it comprise following component:
(1) encapsulates the ELISA Plate of phenolethanolamine A antibody;
(2) phenolethanolamine A horseradish peroxidase-labeled thing working fluid;
(3) phenolethanolamine A standard solution is 6 bottles, and concentration is respectively 0 μ g/L, 0.1 μ g/L, 0.3 μ g/L, 0.9 μ g/L, 2.7 μ g/L, 8.1 μ g/L;
(4) substrate colour developing liquid A liquid is hydrogen peroxide or urea peroxide, and substrate colour developing liquid B liquid is tetramethyl benzidine or OPD;
(5) cleansing solution is the phosphate buffer that contains 0.05% polysorbas20;
(6) the concentrating sample dilution is the phosphate buffer of 0.05% Tween-20;
(7) stop buffer is the hydrochloric acid solution of 2mol/L.
The residual detection of phenolethanolamine A in embodiment 4 samples
1, sample pre-treatments
Accurately take by weighing the 5g feed in the 40ml centrifuge tube.Add 20ml sample diluting liquid (phosphate buffer of 0.01% Tween-20), be heated to 60 ℃, fully vibrated 10 minutes, the centrifugal 10min of 4000rpm gets supernatant 5ml.
2, detect with kit
In the ELISA Plate micropore of phenolethanolamine A antibody sandwich, add series standard article or sample solution 50 μ l, add anti-phenolethanolamine A horseradish peroxidase label 50 μ l again, room temperature reaction 45 minutes.Pour out liquid in the hole, every hole adds 250 μ l through 10 times of cleansing solutions that diluted, and pours out liquid in the hole after 30 seconds, and so repetitive operation is washed plate 3 times altogether, claps with thieving paper and does.Every hole adds substrate colour developing liquid A liquid (urea peroxide or hydrogen peroxide) 50 μ l; Add B liquid (tetramethyl benzidine or OPD) 50 μ l again; The mixing that vibrates gently, room temperature lucifuge colour developing 15min, every hole adds stop buffer (2mol/L hydrochloric acid) 50 μ l; The mixing that vibrates is gently measured every hole absorbance (OD value) with ELIASA.
3, interpretation of result:
Calculate percentage absorbance and drawing standard curve, the concentration of phenolethanolamine A can be read from typical curve in corresponding each sample, also can calculate the content of phenolethanolamine A in sample with regression equation method.Utilize the be more convenient for express-analysis of a large amount of samples of professional computer software.According to the comparison of the depth and the series concentration standard solution color of the sample of color on the ELISA Plate, can judge the concentration range of phenolethanolamine A in the sample.
The test of experimental example 5 kit precision
This experimental example is the repeatable test of standard.Concrete operations are:
In the ELISA Plate of the method preparation from every crowd of embodiment 2 or 3, each extracts 10 holes, measures the absorbance (OD) of the standard solution of 4.5 μ g/L, repeats 3 times, calculates coefficient of variation CV%.
The result shows coefficient of variation scope between 4.8-7.6%, has met the coefficient of variation less than 20% regulation, explains that this kit standard items precision has reached requirement.
The recovery test of experimental example 6 kits
Get the phenolethanolamine A standard specimen of two concentration, sample added recovery test, each concentration do 4 parallel, calculate recovery rate respectively.The result shows that the interpolation recovery in the feed is 60%-80%.
The test of experimental example 3 kit storage lives
The kit preservation condition is 2-8 ℃, and through 6 months mensuration, the maximum absorbance value of kit (zero adds), 50% inhibition concentration, phenolethanolamine A added the practical measurement value all within normal range.Consideration has improper preservation condition and occurs in transportation and use, and kit 37 ℃ of two weeks of preservation condition held, is carried out the accelerated deterioration experiment, and the result shows that this kit each item index meets the requirements fully.Consider that the freezing situation of kit takes place, kit was put into-20 ℃ of refrigerators freezing 5 days, measure the result and show that also kit each item index is normal fully.Can draw kit from above result can preserve more than 12 months at 2-8 ℃.
Claims (8)
1. direct competition method enzyme linked immunological kit that detects phenolethanolamine A, comprising: phenolethanolamine A specific antibody, phenolethanolamine A standard items and phenolethanolamine A horseradish peroxidase-labeled thing.
2. enzyme linked immunological kit according to claim 1 is characterized in that: said kit comprises that also having encapsulated phenolethanolamine A horseradish peroxidase-labeled thing, the ELISA Plate that is coated with phenolethanolamine A specific antibody, phenolethanolamine A standard solution, substrate developer, cleansing solution, stop buffer, concentrating sample dilution and enzyme marks the thing dilution.
3. enzyme linked immunological kit according to claim 1 and 2 is characterized in that: said phenolethanolamine A specific antibody is a polyclonal antibody.
4. enzyme linked immunological kit according to claim 2 is characterized in that: described cleansing solution is for containing 0.05%-0.5% Tween-20 phosphate buffer.
5. enzyme linked immunological kit according to claim 2 is characterized in that: described developer is made up of developer A and developer B, and developer A is hydrogen peroxide or urea peroxide, and developer B is o-phenylenediamine or tetramethyl benzidine.
6. enzyme linked immunological kit according to claim 2 is characterized in that: described concentrating sample dilution is the phosphate buffer that contains 0.1% Tween-20.
7. enzyme linked immunological kit according to claim 2 is characterized in that: described enzyme mark thing dilution is the phosphate buffer that contains 0.01% Tween-20.
8. the method for phenolethanolamine A content in the test sample comprises step:
(1) sample pre-treatments;
(2) detect with the arbitrary described kit of claim 1-7; In the ELISA Plate hole that is coated with phenolethanolamine A antibody, add standard items or sample solution; Add phenolethanolamine A horseradish peroxidase-labeled thing again; Hatch back washing bat and do, add developer, termination, measure absorbance with ELIASA;
(3) analyzing and testing result.
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Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102735834A (en) * | 2012-05-11 | 2012-10-17 | 中国兽医药品监察所 | Enzyme-linked immunoassay kit for detecting phenylethanolamine A |
| CN102901819A (en) * | 2012-09-27 | 2013-01-30 | 江苏维赛科技生物发展有限公司 | Immune colloidal gold detection card for phenylethanolamine A and preparation method thereof |
| CN102901813A (en) * | 2012-09-27 | 2013-01-30 | 江苏维赛科技生物发展有限公司 | Enzyme linked immuno kit for detecting residual phenylethanolamine A and application method thereof |
| CN103265440A (en) * | 2012-12-20 | 2013-08-28 | 苏州大学 | Method of detecting content of phenylethanolamine A and detection kit |
| CN103792226A (en) * | 2012-11-03 | 2014-05-14 | 江苏维赛科技生物发展有限公司 | Chemiluminescence immunodetection kit used for detecting phenylethanolamine A |
| CN104152455A (en) * | 2014-07-30 | 2014-11-19 | 暨南大学 | Phenethylamine aptamer and aptamer electrochemical biosensor for detecting phenethylamine |
| CN105092849A (en) * | 2014-09-16 | 2015-11-25 | 北京勤邦生物技术有限公司 | Test paper strip and method for detecting phenylethanolamine A |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1389730A (en) * | 2002-07-12 | 2003-01-08 | 江南大学 | Clenbterol hydrochloride enzyme immunoassay kit and its immnoassay method |
| CN1707266A (en) * | 2004-06-11 | 2005-12-14 | 中国兽医药品监察所 | Enzyme-linked immunologic kit for detecting clenbuterol |
| CN1766630A (en) * | 2005-11-03 | 2006-05-03 | 北京望尔生物技术有限公司 | ELISA kit for detecting ractopamine in animal derived food |
| CN1920561A (en) * | 2005-08-22 | 2007-02-28 | 上海市上海中学 | Method for preparing antibody for detecting clenbuterol |
| CN101571542A (en) * | 2009-06-15 | 2009-11-04 | 杭州迪恩科技有限公司 | Kit for detecting zilpaterol by direct competitive ELISA (enzyme-linked immunosorbent assay) method |
-
2011
- 2011-12-28 CN CN2011104455178A patent/CN102539747A/en active Pending
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1389730A (en) * | 2002-07-12 | 2003-01-08 | 江南大学 | Clenbterol hydrochloride enzyme immunoassay kit and its immnoassay method |
| CN1707266A (en) * | 2004-06-11 | 2005-12-14 | 中国兽医药品监察所 | Enzyme-linked immunologic kit for detecting clenbuterol |
| CN1920561A (en) * | 2005-08-22 | 2007-02-28 | 上海市上海中学 | Method for preparing antibody for detecting clenbuterol |
| CN1766630A (en) * | 2005-11-03 | 2006-05-03 | 北京望尔生物技术有限公司 | ELISA kit for detecting ractopamine in animal derived food |
| CN101571542A (en) * | 2009-06-15 | 2009-11-04 | 杭州迪恩科技有限公司 | Kit for detecting zilpaterol by direct competitive ELISA (enzyme-linked immunosorbent assay) method |
Cited By (11)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102735834A (en) * | 2012-05-11 | 2012-10-17 | 中国兽医药品监察所 | Enzyme-linked immunoassay kit for detecting phenylethanolamine A |
| CN102735834B (en) * | 2012-05-11 | 2014-09-03 | 中国兽医药品监察所 | Enzyme-linked immunoassay kit for detecting phenylethanolamine A |
| CN102901819A (en) * | 2012-09-27 | 2013-01-30 | 江苏维赛科技生物发展有限公司 | Immune colloidal gold detection card for phenylethanolamine A and preparation method thereof |
| CN102901813A (en) * | 2012-09-27 | 2013-01-30 | 江苏维赛科技生物发展有限公司 | Enzyme linked immuno kit for detecting residual phenylethanolamine A and application method thereof |
| CN102901813B (en) * | 2012-09-27 | 2015-10-28 | 江苏维赛科技生物发展有限公司 | Detect enzyme linked immunological kit and the using method thereof of residual benzene monoethanolamine A |
| CN103792226A (en) * | 2012-11-03 | 2014-05-14 | 江苏维赛科技生物发展有限公司 | Chemiluminescence immunodetection kit used for detecting phenylethanolamine A |
| CN103265440A (en) * | 2012-12-20 | 2013-08-28 | 苏州大学 | Method of detecting content of phenylethanolamine A and detection kit |
| CN103265440B (en) * | 2012-12-20 | 2014-10-22 | 苏州大学 | Method of detecting content of phenylethanolamine A and detection kit |
| CN104152455A (en) * | 2014-07-30 | 2014-11-19 | 暨南大学 | Phenethylamine aptamer and aptamer electrochemical biosensor for detecting phenethylamine |
| CN104152455B (en) * | 2014-07-30 | 2016-07-06 | 暨南大学 | The aptamers electrochemica biological sensor of gram rumba amine aptamers and detection Ke Lunba amine |
| CN105092849A (en) * | 2014-09-16 | 2015-11-25 | 北京勤邦生物技术有限公司 | Test paper strip and method for detecting phenylethanolamine A |
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