CN104215754B - The eukaryotic expression of SLO albumen and containing the kit of eukaryotic expression SLO albumen - Google Patents
The eukaryotic expression of SLO albumen and containing the kit of eukaryotic expression SLO albumen Download PDFInfo
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Abstract
The invention provides a kind of eukaryotic expression of SLO albumen and the kit containing eukaryotic expression SLO albumen. The present invention obtains SLO albumen by eukaryotic expression, also utilize this albumen to invent a kind of kit of the ASO of detection content, described kit is made up of R1 reagent, R2 reagent and calibration object three parts, wherein R1 reagent is phosphate buffer, R2 reagent is latex suspension, and calibration object is mainly cow's serum matrix. It is high, active strong that the present invention adopts eukaryotic expression system to give expression to purity, and adopt the coated SLO proteantigen of composite emulsion particle, solve in the market that ASO detection sensitivity is not high, repeatability is not good enough, can not meet the clinical problem to the quantitative requirement of ASO, obviously improved sensitivity, accuracy and the linearity of measurement result.
Description
Technical field
The invention belongs to medical immunology in-vitro diagnosis field, be specifically related to eukaryotic expression and the application of SLO albumen, particularly onePlant the eukaryotic expression of SLO albumen and the kit containing eukaryotic expression SLO albumen.
Background technology
SLO (streptococcus blood lysin O) is one of metabolite of A group of streptococcus, and it can cause HRBC's dissolving,There is very strong antigenicity. People is subject to 2-3 week after hemolytic streptococcal infection, just produces antistreptolysin O (ASO) in bodyAntibody, i.e. antistreptolysin O (ASO) (ASO).
A group of streptococcus can cause various infection, and these infection can cause heart and kidney damage. By early diagnosis, haveEffect treatment, and detection to patient, can reduce these risks. ASO measures for the streptococcal infection of diagnosis A family valency very muchValue, its existence and content can reflect the order of severity of infection. A group of streptococcus infects latter 1 week, and ASO starts to raise, 4～Within 6 weeks, can reach peak, and can continue the several months, in the time that infection is gone down, the decline of ASO value was also got back to normal value in 6 months,If ASO titre does not decline, may there is recurrent infection or chronic infection in hint. Repeatedly measure, antibody titer graduallyRaise significant to diagnosing, antibody titer declines gradually, and state of an illness alleviation is described. Rheumatic fever, acute glomerulonephritis,The ASO such as erythema nodosum, scarlet fever, acute tonsillitis obviously raise. Minority hepatitis, CTD, tuberculosisAnd myelomatosis multiplex people also can make ASO increase. Because people often contact with A family streptococcus, normal person also exists lowThe antibody of tiring, conventionally, when tiring > when 200U/ml, be just considered to diagnostic value. Be less than the Healthy People of 15-20%ASO content in serum is higher than 200U/ml. Most of neonatal ASO content is higher than its mother, but after its birthIn several weeks, ASO content can sharply decline. Preschool child's ASO value is usually less than 100U/ml, then with the increasing at ageAdding ASO value increases, and reaches peak in school age, and after growing up, ASO value declines. Except acute phase, rheumatoid disease is closedSave the rising that conventionally can't detect ASO value in scorching patient's serum. Nephrotic syndrome and syndrome of antibody deficiency patientIn serum, only there is the ASO of utmost point low content.
ASO detects principle: the table that with unique technology, antigen (streptolysin " O ") is coated on to trickle latex particleFace, this latex particle that has been coated with antigen meets with the ASO detecting in sample serum, forms antigen-antibody complex,Generation turbidity changes. And strengthen turbidity through the participation of latex particle and change, thereby quantitative detection low concentration that can be responsiveASO. By measuring 600nm wavelength absorbance, and with calibration curve comparison, obtain the content of ASO in sample.
Current domestic detection ASO generally adopts latex agglutination, although the method is easy and simple to handle, and its repeatability, stableProperty etc. is poor, and sensitivity is not high, especially can only carry out qualitative results analysis, can not meet the clinical requirement quantitative to ASO.Reason is that domestic ASO detects reagent place's antigen and has the problems such as purity is not high, difference between batch is large, and major antigen sourceMain dependence import, expensive. The present invention adopts composite-grain diameter latex particle to strengthen immunoturbidimetry and detects in serumASO content. The SLO albumen that adopt the purity of autonomous preparation high, activity is strong, and adopt compounded latex particle coated anti-Former, make method sensitivity of the present invention can detect that 30U/ml improves, linearity reaches 600U/ml.
Summary of the invention
For defect of the prior art, the object of this invention is to provide a kind of eukaryotic expression of SLO albumen and contain eucaryonExpress the kit of SLO albumen. The invention provides a kind of method of eukaryotic expression SLO albumen, utilize the method to be somebody's turn to do simultaneouslySLO albumen prepared by method, the test kit of having invented a kind of composite-grain diameter latex particle envelope antigen, this kitFor detection of the ASO content in serum, reaching easy and simple to handle, highly sensitive, specificity is good, fast, measurement resultObject accurately and reliably.
The present invention is achieved by the following technical solutions:
First aspect, the present invention relates to a kind of method of utilizing methanol yeast system eukaryotic expression SLO albumen, specifically comprisesFollowing steps:
(1) adopt eukaryotic expression system, SLO Gene cloning, in the MCS of expression vector, is obtained to recombinant vector;Described eukaryotic expression system be high-fidelity methanol yeast bacterium (PichiaPastoris) expression system (LeiJ, GuanB,LiB，eta1.Expression，purificationandcharacterizationofrecombinanthumaninterleukin-2-serumalbumin(rhIL-2-HSA)fusionproteininPichiapastoris)；
(2) recombinant vector extracts recombinant plasmid after being transformed into a large amount of amplifications of host cell; Described host cell is large intestine barBacterium competence cell;
(3) recombinant plasmid linearisation; Described linearisation is to carry out restriction enzyme digestion by SalI;
(4) then with electric shock conversion method, linearizing plasmid vector is proceeded to saccharomycete;
(5) saccharomycete is first grown in glycerinated culture medium, is cultured to after high concentration, lures taking methyl alcohol as sole carbon sourceLead and express external source SLO albumen.
Preferably, in described step (1), expression vector comprises pPIC9K, pDBLeu, pEXP-AD502 or pPC-86In one.
Second aspect, the present invention relates to a kind of kit containing eukaryotic expression SLO albumen, comprises R1 reagent, R2 reagentAnd calibration object, described R1 reagent is phosphate buffer, described R2 reagent is the sensitization latex that has been cross-linked SLO antigenSuspension, described calibration object comprises all identical cow's serum matrix of 4 ASO concentration differences, solutes content.
Preferably, in described phosphate buffer, solvent is pH7.0～7.6,30～80mmol/L phosphate-bufferedLiquid, solute and the content in described phosphate buffer thereof are: Macrogol 200-1000060～120mmol/L,Disodium ethylene diamine tetraacetate 10～20mmol/L.
Preferably, the described sensitization latex suspension that is cross-linked SLO antigen, solvent be pH7.0～7.6,40～The phosphate buffer of 100mmol/L, suspended particulate and the solute content in described suspension is: composite S LO sensitizationParticle 0.1～1%, disodium ethylene diamine tetraacetate 5～25mmol/L, skimmed milk power 0.05～1%, NaN30.2～1.5％。
Preferably, being prepared as of described composite S LO sensitization particle: the 20mmol/L, the pH7.4 that contain SLO albumenPhosphate buffer and polystyrene latex mix with the mass ratio of 30:100,37 DEG C of absorption 8 hours, dialysis is removed notSLO albumen in connection, adds confining liquid, seals 2 hours, centrifugally removes supernatant and get final product; Described confining liquid is for containing 0.1%The glycine of (mass percent) skimmed milk power, 0.2mmol/L; The preparation of described composite S LO sensitization particle adopts thingReason absorption combines with chemical crosslinking and realizes the crosslinked of streptolysin O antigen and latex particle.
Preferably, described ASO concentration is respectively 75,150,300 and 600U/mL.
Preferably, in described calibration object cow's serum matrix, solute and content thereof are: anticorrisive agent 0.2～2.2%, Tween-20 1～10%, protective agent 1～3%, ASO.
Preferably, anticorrisive agent described in described calibration object is Sodium azide; Described protective agent comprises glycerine, sucrose, cow's serumOne or more in albumin (BSA), sweet mellow wine, sorbierite.
Compared with prior art, the present invention has following beneficial effect:
(1) kit calibration object quality-control product of the present invention adopts cow's serum matrix, this cow's serum matrix by without HBV, HIV,HAV, HCV, the TP infection sources, endanger minimum to operator;
(2) it is anti-that the coating technique that R2 reagent adopts physical absorption to combine with chemical crosslinking has been realized streptolysin OFormer and latex particle crosslinked, the stability of compound strengthens greatly, is conducive to improve 37 DEG C of stability of reagent and effect phaseStability;
(3) company independently adopts methanol yeast system can give expression in a large number, the SLO antigen that purity is high, thereby as far as possibleThe false positive of having avoided ASO testing result, the reagent test degree of accuracy improves greatly, the antigen of expression can replace abroadImport SLO antigen;
(4) detection kit of the present invention, by the optimization of component and content, makes to detect linear wide ranges, dense at sampleDegree still can detect result while being low to moderate 30U/ml, and the minimum detected value of commercial reagent (as silent happy biological reagent) is 100U/ml；
(5) kit of the present invention obviously improves with commercial reagent (silent happy biological reagent) phase specific sensitivity, as the present inventionWhen kit measurement physiological saline, absorbance is changed to 0.58 (1/10000A), and theoretical concentration is 100U/ml serum sampleTime absorbance be changed to 1263.2 (1/10000A), and silent happy biological reagent while measuring physiological saline absorbance be changed to-28(1/10000A) when, theoretical concentration is 100U/ml serum sample, absorbance is changed to 523.8 (1/10000A);
(6) kit of the present invention is compared with commercial reagent (silent happy biological reagent), and the degree of accuracy is obviously high, as the present inventionReagent is measured the basic, normal, high value determination of serum of Bole's specific protein Quality Control mean value and is respectively 81.31U/mL, 128.77U/mLAnd 175.70U/mL, be respectively-2.27%, 1.39% and 3.35% with the relative deviation of target value; Silent happy biological reagent is measuredMean value is respectively 77.00U/mL, 122.99U/mL and 161.77U/mL, is respectively with the relative deviation of target value-7.45% ,-3.16% and-4.84%.
Brief description of the drawings
By reading the detailed description of non-limiting example being done with reference to the following drawings, other spy of the present inventionLevy, object and advantage will become more obvious:
Fig. 1 is the calibration curve of the ASO normative reference of 4 kinds of different contents;
Wherein, each point represents the normative reference of different content ASO, and X-axis represents the content of ASO, and Y-axis represents extinctionDegree;
Fig. 2 is the ASO latex enhancement mode reagent that adopts respectively reagent of the present invention and Beckman company of the U.S., adopts dayVertical automatic clinical chemistry analyzer carries out by each autoregressive parameter 50 parts of human serums (comprising normal and monstrosity) simultaneouslyMeasure, measured value is carried out to correlation analysis. What wherein X-axis represented is patients serum's result that reagent of the present invention is measured,What Y-axis represented is patients serum's result that Beckman company of U.S. reagent is measured, coefficient R2=0.98, regression equationFor y=0.943x+14.78.
Detailed description of the invention
Below in conjunction with specific embodiment, the present invention is described in detail. Following examples will contribute to the technology people of this areaMember further understands the present invention, but does not limit in any form the present invention. It should be pointed out that the common skill to this areaArt personnel, without departing from the inventive concept of the premise, can also make some distortion and improvement. These all belong toProtection scope of the present invention.
Embodiment 1
The present embodiment relates to a kind of eukaryotic expression of SLO albumen, comprises the steps:
1. the structure of carrier for expression of eukaryon: obtain SLO genetic fragment by PCR method, be connected to T carrier,Cut rear recovery gene segment with EcoRI and SalI enzyme, cut carrier for expression of eukaryon pPIC9K with EcoRI and SalI enzymeRear recovery carrier segments, carries out coupled reaction by this two fragment through DNA ligase, connects product transformed competence colibacillus cellBL21, picking monoclonal overnight incubation, extracts plasmid, cuts qualification through EcoRI and SalI enzyme, to obtaining expection sizeThe carrier of Insert Fragment check order. The carrier called after pPIC9K-SLO that checks order correct.
2. recombinant vector is transformed in competent escherichia coli cell, selects positive colony, expand after cultivation, use matterGrain drawer kit extracts required plasmid, then carries out restriction enzyme digestion by Sac II, makes it total Linearization, 1% agarAfter sugar electrophoresis, reclaiming kit recovery linearization plasmid with glue is dissolved in 1 × TE.
3. the linearisation DNA of 5～20 μ g is added in competence yeast strain, be transferred to pole cup, put 5 minutes on ice;With the Bio-RadGenePulser conversion of shocking by electricity, add immediately the 1M sorbierite of precooling, be coated with dull and stereotypedly, put 28 DEG C of trainingsSupporting 48h occurs to transformant.
4. screening picking list bacterium colony, is inoculated into fluid nutrient medium (BMGY/BMMY culture medium), cultivates after 48 hours, uses100% methanol induction is expressed SLO albumen, to the final concentration of methyl alcohol in culture medium be 0.8%.
Embodiment 2
The present embodiment provides a kind of kit of the SLO of containing eukaryotic expression albumen, composed as follows:
1. reagent R1 is:
| PH7.4 phosphate buffer | 30 mmol/L |
| Macrogol 200 | 60 mmol/L |
| Disodium ethylene diamine tetraacetate | 10 mmol/L |
2. reagent R2 is:
Streptolysin O and polystyrene latex mix the (phosphorus of buffer solution employing 0.02M with the mass ratio of 30:100Acid buffer (pH7.4), mixes rear 37 DEG C of absorption 8 hours by both, and the antigen not connecting is removed in dialysis afterwards, addsEnter confining liquid (glycine buffer of 0.1% skimmed milk power, 0.2mm), seal 2 hours. The centrifugal supernatant that goes, uses latexDilution (pH7.4 phosphate buffer 50mmol/l, disodium ethylene diamine tetraacetate 15mmol/L, 0.1% skimmed milk power,0.9％NaN3) to 0.2%.
3. cow's serum matrix calibration object preparation:
By treated cow's serum, add BSA (bovine serum albumin) 1%, NaN30.9%, Tween-20 1.5% obtainsCalibration object dilution. Be dissolved in the solution of the similar human serum matrix of preparation with ASO, prepare the calibration object (75 of variable concentrationsU/ml，150U/ml，300U/ml，600U/mL)。
The ASO detection kit that the present embodiment is described, is applicable to various types of full automatic biochemical apparatus, with Hitachi 7170Full automatic biochemical apparatus is example, and it operates as table 1. Analytical method: Two point end assay, the consumption of reagent R1, R2 respectivelyFor 280ul and 70ul, sample size 6ul; 280ul reagent R1 adds 6ul sample to add 70ulR2 after 37 DEG C of 5min,Start read point, after reaction 5min, read another point; Detection dominant wavelength is 600nm.
Adopt this reagent and said determination method, the ASO of 4 kinds of different contents that employing Hitachi 7170 Biochemical Analyzers recordThe curve (as shown in Figure 1) of calibration object (self-control), each point represents the reference calibrations product of a content, wherein X-axisRepresent ASO content (U/ml); Y-axis represents absorbance.
Table 1
Embodiment 3: the correlation test that detects reagent
Use the ASO glue of this law invention kit (specifically filling a prescription with embodiment 1) and Beckman company of the contrast agents U.S.Breast enhancement mode reagent, adopts automatic 7170 automatic clinical chemistry analyzers to 50 parts of human serums (comprising normal and monstrosity)Measure by each autoregressive parameter simultaneously, measured value is carried out to correlation analysis. According to above-mentioned " table 1 " in parameter enterRow is measured measurement result and is seen Fig. 2, X, and Y-axis is measured value (the content U/ml of ASO),
Result by Fig. 2 finds out, the relevant of two kinds of reagent is R2=0.98, and regression equation is y=0.9435x+14.781.It is good that result shows that this reagent and import reagent are measured patients serum's correlation, has good specificity and accuracy. ThisOutward, above experiment is that 7170 full automatic biochemical apparatus that adopt Hitachi, Ltd to manufacture carry out, but reagent of the present invention is not limitIn above-mentioned instrument, be also applicable to other full-automatic or semi-automatic biochemical analyzers.
Test example 4: LDL test
This experiment purpose is to detect the minimum check-up inducing degree of reagent in the time of test clinical sample.
Adopt experimental example 1 reagent, contrast agents (Jiangsu is silent happy biological), calibration object, blank solution (normal saline solution),Normal human serum sample, low value sample (sample of numerical value in reagent range of linearity lower limit ± 1/3).
Machine: Hitachi's 7170 automatic biochemistry analyzers.
Operating procedure: use normal saline solution or deionized water dissolving low value sample, then 50% be diluted to 5 points,Each test sample 5 times together with zero point, calculating mean value, tries to achieve SD numerical value.
Result is resolved: according to detecting data, calculate SD numerical value and CV numerical value, calculate respectively 1SD, 2SD, from minimumBeginning, the numerical value of its mean value-2SD is exactly the minimum check-up inducing degree of reagent more than zero point mean value+2SD.Table 2 shows, when kit reagent of the present invention is measured dilution 1/16,1/8,1/4,1/2 serum, and wherein 1/4,1/2The numerical value of dilution mean value-2SD is greater than mean value+2SD at zero point, shows that kit reagent LDL of the present invention is passableReach 30U/ml. Table 3 shows, silent happy biological reagent mensuration dilution 1/16,1/8,1/4,1/2 serum, and compare bloodPeaceful average-2SD and zero point mean value+2SD size, it is average that the numerical value of all dilution mean value-2SD is all less than zero pointValue+2SD, shows that silent happy biological reagent lowest detection is limited to 100U/ml left and right.
Table 2
Table 3
Test example 5: sensitivity experiment
This experiment purpose is that the absorbance of kit reagent in the time of test physiological saline and certain density management serum becomesChange value.
Adopt experimental example 1 reagent, contrast agents (Jiangsu is silent happy biological), calibration object, blank solution, 0.9% physiologySaline solution, the absorbance changing value when management serum of concentration.
Machine: Hitachi's 7170 automatic biochemistry analyzers.
Operating procedure: use normal saline solution, low value sample, each test sample 5 times, calculates absorbance.
Table 4,5 demonstrations, when reagent of the present invention is measured physiological saline, absorbance is changed to 0.58 (1/10000A), theoretical denseWhen degree is 100U/ml serum sample, absorbance is changed to 1263.2 (1/10000A), and silent happy biological reagent is measured physiologyWhen salt solution, absorbance is changed to-28 (1/10000A), and when theoretical concentration is 100U/ml serum sample, absorbance is changed to523.8 (1/10000A), show that kit reagent sensitivity of the present invention will be significantly higher than silent happy biological reagent. Table 4,5Represent respectively invention kit reagent and the sensitivity of Mo Le biological reagent, show that kit reagent sensitivity of the present invention is obviously goodIn silent happy biological reagent.
Table 4
Table 5
Embodiment 6: degree of accuracy experiment
This experiment purpose is to detect the reagent degree of accuracy.
Operating procedure: test respectively 3 basic, normal, high values of Bole's specific protein Quality Control serum (lot number 52480), calculateAverage and with the relative deviation of quality-control product target value.
Result is resolved: according to detecting data, calculate relative deviation, the absolute value of deviation is less, represents that the degree of accuracy is higher.The demonstration of table 6 result, reagent of the present invention is measured the basic, normal, high value determination of serum of Bole's specific protein Quality Control mean value and is respectively81.31U/mL, 128.77U/mL and 175.70U/mL, be respectively-2.27%, 1.39% and with the relative deviation of target value3.35%; Silent happy biological reagent is measured mean value and is respectively 77.00U/mL, 122.99U/mL and 161.77U/mL, withThe relative deviation of target value is respectively-7.45% ,-3.16% and-4.84%. The degree of accuracy that shows reagent of the present invention is high.
Table 6
Embodiment 7
The present embodiment relates to a kind of eukaryotic expression of SLO albumen, and concrete steps are identical with embodiment 1, and difference onlyBe, carrier for expression of eukaryon is pEXP-AD502.
Embodiment 8
The present embodiment relates to a kind of eukaryotic expression of SLO albumen, and concrete steps are identical with embodiment 1, and difference onlyBe, carrier for expression of eukaryon is pDBLeu.
Embodiment 9
The present embodiment provides a kind of kit of the SLO of containing eukaryotic expression albumen, and technical scheme is identical with embodiment 2, instituteIf different component and content are as follows:
Reagent R1:
| PH7.4 phosphate buffer | 50 mmol/L |
| PEG20000 | 90 mmol/L |
| Disodium ethylene diamine tetraacetate | 15 mmol/L |
Reagent R2: composite S LO sensitization particle 0.1%, disodium ethylene diamine tetraacetate 5mmol/L, skimmed milk power 0.05%,NaN30.2％。
Calibration object: NaN30.2%, sorbierite 2%, Tween-20 5%.
Embodiment 10
The present embodiment provides a kind of kit of the SLO of containing eukaryotic expression albumen, and technical scheme is identical with embodiment 2, instituteIf different component and content are as follows:
Reagent R1:
| PH7.6 phosphate buffer | 80 mmol/L |
| Macrogol 4000 | 120 mmol/L |
| Disodium ethylene diamine tetraacetate | 20 mmol/L |
Reagent R2: composite S LO sensitization particle 1%, disodium ethylene diamine tetraacetate 25mmol/L, skimmed milk power 1%,NaN31.5％。
Calibration object: NaN32.2%, sucrose 2%, Tween-20 10%.
Above specific embodiments of the invention are described. It will be appreciated that, the present invention is not limited toState specific implementations, those skilled in the art can make various distortion or amendment within the scope of the claims,This does not affect flesh and blood of the present invention.
Claims (5)
1. the kit containing eukaryotic expression SLO albumen, it is characterized in that, comprise R1 reagent, R2 reagent and calibration object, described R1 reagent is phosphate buffer, described R2 reagent is the sensitization latex suspension that has been cross-linked SLO antigen, and described calibration object comprises all identical cow's serum matrix of 4 different ASO concentration, solutes content;
The eukaryotic expression of described SLO antigen, specifically comprises the steps:
(1) adopt eukaryotic expression system, SLO Gene cloning, in the MCS of expression vector, is obtained to recombinant vector; Described eukaryotic expression system is the methanol yeast bacterium expression system of high-fidelity;
(2) recombinant vector extracts recombinant plasmid after being transformed into a large amount of amplifications of host cell; Described host cell is competent escherichia coli cell;
(3) recombinant plasmid linearisation; Described linearisation is to carry out restriction enzyme digestion by SalI;
(4) then with electric shock conversion method, linearizing recombinant plasmid is proceeded to saccharomycete;
(5) saccharomycete step (4) being obtained is first grown in glycerinated culture medium, is cultured to after high concentration, taking methyl alcohol as sole carbon source abduction delivering external source SLO albumen;
In described step (1), expression vector comprises pPIC9K, pDBLeu, pEXP-AD502 or pPC-86;
The solvent of the described sensitization latex suspension that has been cross-linked SLO antigen is the phosphate buffer of pH7.0～7.6,40～100mmol/L, suspended particulate and the solute content in described suspension is: composite S LO sensitization particle 0.1～1%, disodium ethylene diamine tetraacetate 5～25mmol/L, skimmed milk power 0.05～1%, NaN30.2～1.5％；
Being prepared as of described composite S LO sensitization particle: the 20mmol/L that contains SLO albumen, the phosphate buffer of pH7.4 and polystyrene latex mix with the mass ratio of 30:100,37 DEG C adsorb 8 hours, the SLO albumen not connecting is removed in dialysis, add confining liquid, seal 2 hours, centrifugally remove supernatant and get final product; The glycine that described confining liquid contains 0.1% skimmed milk power, 0.2mmol/L; The preparation of described composite S LO sensitization particle adopts physical absorption to combine with chemical crosslinking and realizes the crosslinked of streptolysin O antigen and latex particle.
2. the kit containing eukaryotic expression SLO albumen according to claim 1, it is characterized in that, in described phosphate buffer, solvent is pH7.0～7.6,30～80mmol/L phosphate buffer, solute and the content in described phosphate buffer thereof are: Macrogol 200-1000060～120mmol/L, disodium ethylene diamine tetraacetate 10～20mmol/L.
3. the kit containing eukaryotic expression SLO albumen according to claim 1, is characterized in that, described ASO concentration is respectively 75,150,300 and 600U/mL.
4. the kit containing eukaryotic expression SLO albumen according to claim 1, is characterized in that, in described calibration object cow's serum matrix, solute and content thereof are: anticorrisive agent 0.2～2.2%, Tween-20 1～10%, protective agent 1～3%, ASO.
5. the kit containing eukaryotic expression SLO albumen according to claim 4, is characterized in that, described anticorrisive agent comprises Sodium azide; Described protective agent comprises one or more in glycerine, sucrose, BSA, sweet mellow wine, sorbierite.
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| CN104483492A (en) * | 2014-12-22 | 2015-04-01 | 宁波美康生物科技股份有限公司 | Detection kit for anti-streptolysin O antibody |
| CN104865379B (en) * | 2015-05-02 | 2017-06-13 | 王贤俊 | A kind of antistreptolysin O detection method |
| CN104931706B (en) * | 2015-05-02 | 2017-06-13 | 王贤俊 | A kind of antistreptolysin O detection kit and its preparation |
| CN106153887A (en) * | 2016-05-27 | 2016-11-23 | 安徽伊普诺康生物技术股份有限公司 | A kind of kit measuring antistreptolysin O |
| CN107167586B (en) * | 2017-07-06 | 2019-03-26 | 四川新健康成生物股份有限公司 | A kind of antistreptolysin O (ASO) detection kit and preparation method thereof |
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