CN110308282A - A kind of stable homocysteine Enzymatic cycling detection kit - Google Patents
A kind of stable homocysteine Enzymatic cycling detection kit Download PDFInfo
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Abstract
The present invention provides a kind of stable homocysteine Enzymatic cycling detection kit; the kit includes reagent R1 and R2; wherein reagent R1 is by buffer; surfactant, protective agent, antioxidant; disodium ethylene diamine tetraacetate; three (2- carbonylethyl) microcosmic salt hydrochlorates, S-adenosylmethionine, α-ketoglutaric acid; reduced coenzyme and preservative composition; reagent R2 is by buffer, surfactant, protective agent; Adenosylhomocysteinase EC3.3.1.1; Hcy transmethylase, adenosine deaminase, glutamte dehydrogenase and preservative composition.Synergistic effect of the kit provided by the invention due to surfactant and protective agent, antioxidant; it is set to compare commercially available HCY kit; have many advantages, such as that good precision, high sensitivity, the range of linearity are wide and stability is good, the auxiliary diagnosis for clinic HCY related disease provides important evidence.
Description
Technical field
The present invention relates to biochemical diagnosis reagent preparation fields, follow specifically, being related to a kind of stable homocysteine
Ring enzyme process detection kit.
Background technique
Homocysteine (HCY) is also known as homocysteine, is a kind of amino acid containing sulfydryl, belongs to methionine and half Guang
The important intermediate generated in propylhomoserin metabolic process.HCY exists in two forms in human plasma, half Guang of oxidized form homotype
Propylhomoserin and reduced form homocysteine.The sulfydryl to dissociate in reduced form homocysteine has very high activity, is easy
It is oxidized and forms disulfide form；Oxidized form is that homocysteine is primarily present form in blood plasma, with curing
Object form exists with protein bound form.Under normal circumstances, homocysteine can be decomposed metabolism in vivo in blood,
Concentration maintains reduced levels.But under the influence of primary and secondary disease, homocysteine concentration is caused to increase,
The HCY of high concentration can be such that oxyradical largely generates and cause endothelial cell damage, make the low close of lipid peroxidation and oxidized form
Degree increases, to cause the damage and destruction of blood vessel, damaged vascular wall, which increases, causes patch, causes angiocarpy, artery athero-
The diseases such as hardening and apoplexy.A large number of studies show that hyperhomocysteinemiainjury atherosclerosis and thrombotic diseases such as
It plays an important role in the pathogenesis such as cranial vascular disease, coronary heart disease and phlebothrombosis.
The measuring method of homocysteine mainly has high performance liquid chromatography (HPLC method), gas chromatography-mass spectrography
(GC-MS method), ImBaunofenzymatic technique (ELA method), fluorescence polarization immunoassay method (FPLA method) and Enzymatic cycling.Chromatography has
Very high accuracy and stability, however, it is more demanding to the handling and operation personnel of sample, plant maintenance, usually as same
The reference method of type cysteine, and cannot be used for the detection of conventional sample；The high sensitivity of enzyme-linked immunization, high specificity,
But it is cumbersome, precision is poor, and consuming time is long, is not suitable for the detection of great amount of samples；Fluorescence polarization immunoassay method, needs
It is equipped with dedicated detecting instrument, single testing cost is higher；Enzymatic cycling can be used for large-scale Biochemical Analyzer, easy to operate, essence
Density is good, is suitble to clinical detection at present.
Enzymatic cycling includes two methods of hydrolysis enzyme process and cystathionie method at present.Cystathionie method is because that cannot eliminate eubolism
β-cystathionie, extra high β-cystathionie can be generated particularly with renal disease patient, so as to cause the inaccurate of testing result
Really.Enzyme process is hydrolyzed, specificity is high, strong antijamming capability, and it is more accurate to eliminate metabolism β-cystathionie, testing result in vivo.City
It is mostly offshore company's monopolization that enzymatic reagent is hydrolyzed on field, causes high testing cost, therefore, needs to develop a kind of anti-interference ability
By force, accuracy is high, low-cost HCY detection reagent.
Summary of the invention
The object of the present invention is to provide a kind of stable homocysteine Enzymatic cycling detection kits.
In order to achieve the object of the present invention, the present invention provides the detection examination of stable homocysteine (double reagent) Enzymatic cycling
Agent box, the kit include reagent R1 and R2, for the liquid double reagent used on Biochemical Analyzer；
The formula of the reagent R1 are as follows: the buffer of 20-200mM, pH7.5~9.5, surfactant 0.5-5g/L are protected
Protect agent 0.5-20g/L, antioxidant 0.05-50mM, disodium ethylene diamine tetraacetate (EDTA-2Na) 0.2-20mM, three (2- carbonyls
Ethyl) microcosmic salt hydrochlorate (TCEP) 0.2-10mM, S-adenosylmethionine (SAM) 0.05-5mM, α-ketoglutaric acid (α-KG) 1-
20mM, NADH (reduced Coenzyme I) 0.1-10mM, preservative 0.5-2g/L；
The formula of the reagent R2 are as follows: the buffer of 20-200mM, pH7.5~9.5, surfactant 0.5-20g/L are protected
Protect agent 0.5-5g/L, Adenosylhomocysteinase EC3.3.1.1 (SAHase) 1-10KU/L, Hcy transmethylase (HMTase) 2-
20KU/L, adenosine deaminase (ADA) 1-10KU/L, glutamte dehydrogenase (GLDH) 1-20KU/L, preservative 0.5-2g/L；
Wherein, the buffer be selected from borate buffer, Triethanolamine buffer, Tris buffer, MES buffer,
At least one of HEPES buffer solution, PIPES buffer, CAPSO-Na buffer, PBS buffer solution etc..
The surfactant is selected from Tween-20, Triton X-100, TX-10, ON-870, F68, Brij35,16
At least one of alkyl ammomium chloride etc..
The protective agent is selected from least one of BSA, sucrose, lactose, trehalose, D-sorbite, mannitol etc..
The antioxidant is selected from ascorbic acid, tert-butyl hydroquinone, thio-2 acid, butylated hydroxy anisole, two
At least one of butylated hydroxytoluene etc..
The preservative is NaN3 and/or Proclin-300.
Detection kit of the invention further includes HCY calibration object solution, and the concentration of the HCY calibration object solution is respectively 2 μ
M,8μM,15μM,30μM,60μM；Reagent for preparing the HCY calibration object solution is BSA containing 10g/L and 1g/L
The 50mM PBS buffer solution of Proclin-300.
In the specific embodiment of the present invention, the detection kit is by reagent R1, R2 and HCY calibration object solution
Composition；
The formula of the reagent R1 are as follows: the Tris buffer of 50mM, pH8.5, Triton X-100 1g/L, mannitol 5g/
L, tert-butyl hydroquinone 1mM, disodium ethylene diamine tetraacetate 0.5mM, three (2- carbonylethyl) microcosmic salt hydrochlorate 2mM, S- adenosine first
Methyllanthionine 1mM, α-ketoglutaric acid 5mM, NADH 0.4mM, NaN31g/L；
The formula of the reagent R2 are as follows: the HEPES buffer solution of 50mM, pH9.0, Triton X-100 1g/L, mannitol
5g/L, Adenosylhomocysteinase EC3.3.1.1 3KU/L, Hcy transmethylase 5KU/L, adenosine deaminase 2KU/L, glutamic acid
Dehydrogenase 10 KU/L, NaN31g/L；
Concentration is respectively 2 μM, 8 μM, 15 μM, 30 μM, 60 μM of HCY calibration object solution.
Detection kit of the invention can be prepared as follows to obtain:
Reagent preparation R1: weighing each component in proportion, first prepares 20-200mM buffer, and stirring and dissolving adjusts pH value of solution
Value is 7.5~9.5, sequentially adds surfactant, protective agent, antioxidant, disodium ethylene diamine tetraacetate, three (2- carbonyls
Ethyl) microcosmic salt hydrochlorate, S-adenosylmethionine, α-ketoglutaric acid, reduced Coenzyme I and preservative, it mixes, after stirring and dissolving,
It is settled to preformulation volume with purified water, with 0.45 μm of membrane filtration, after mixing, is stored in 2 DEG C~8 DEG C；
Reagent preparation R2: weighing each component in proportion, first prepares 20-200mM buffer, and stirring and dissolving adjusts pH value of solution
Value is 7.5~9.5, sequentially adds surfactant, protective agent, Adenosylhomocysteinase EC3.3.1.1, the transfer of Hcy methyl
Enzyme, adenosine deaminase, glutamte dehydrogenase and preservative mix, and after stirring and dissolving, are settled to preformulation volume with purified water,
With 0.45 μm of membrane filtration, after mixing, stored in 2 DEG C~8 DEG C；
It prepares calibration object solution: first preparing the 50mM PBS buffer solution of BSA containing 10g/L and 1g/L Proclin-300, with
It for preparation of reagents concentration is respectively 2 μM, 8 μM, 15 μM, 30 μM, 60 μM of HCY calibration object solution.
By the above-mentioned reagent prepared, dispensed according to 60mL/ bottles and 20mL/ bottles of reagent R2 of ratio of R1；HCY
Calibration object solution is dispensed according to 1mL/ branch, then by after packing reagent and calibration object label mounted box, as half Guang ammonia of homotype
Sour Enzymatic cycling detection kit.
The testing principle of detection kit of the present invention is as follows: work of the sample in three (2- carbonylethyl) microcosmic salt hydrochlorates (TCEP)
Under, oxidized form homocysteine is converted into sequestered Hcy, and sequestered Hcy and S-adenosylmethionine (SAM) are in Hcy
Reaction generates methionine and AdoHcy (SAH) under the catalysis of transmethylase (HMTase).SAH is by S- adenosine
Homocysteine hydrolase (SAHase) is hydrolyzed into adenosine and Hcy, and the Hcy of formation can recycle addition reaction (half Guang of homotype
The amount of propylhomoserin is amplified by circulation), to be exaggerated detection signal.It is further coupled the dehydrogenation reaction of NADH simultaneously.Product adenosine
It is soon hydrolyzed to hypoxanthine and ammonia, ammonia makes NADH be converted into NAD+, sample under the action of glutamte dehydrogenase (GLDH)
In Hcy content it is directly proportional to the variation of NADH.Under 340nm wavelength, by being compareed with the calibration object equally handled, determine
The content of HCY in sample.
By above-mentioned technical proposal, the present invention at least have following advantages and the utility model has the advantages that
(1) homocysteine detection kit provided by the invention, detection is quickly, the range of linearity is wide, accuracy is high,
Sensitivity is good, low in cost.For Monitoring lower-cut up to 2 μm of ol/L, the range of linearity is 2~60 μm of ol/L, for clinical Accurate Diagnosis
It provides safeguard.
(2) detection kit provided by the invention is made prepared due to the synergistic effect of protective agent and antioxidant
Kit has preferable stability: 2~8 DEG C of kits save 12 months, and accuracy in detection is still good；Reagent is uncapped stationary phase
Up to 14 days, the extension for stability of uncapping saved the time and efforts that clinical examination operator frequently replaces reagent.
(3) content of HCY in sample can be directly measured using detection kit of the invention, it is anti-interference compared to indirect method
Ability is strong, and the interference vulnerable to β-cystathionie of eubolism and other detection reagents, measurement result be not more accurate for measurement result.
Detailed description of the invention
Fig. 1 is the linear detection range test result of homocysteine detection kit in the embodiment of the present invention 3.
Specific embodiment
The following examples are used to illustrate the present invention, but are not intended to limit the scope of the present invention..Unless otherwise specified, embodiment
Used in the conventional means that are well known to those skilled in the art of technological means, raw materials used is commercial goods.
The preparation of 1 homocysteine of embodiment (double reagent) Enzymatic cycling detection kit
Homocysteine (double reagent) Enzymatic cycling detection kit provided in this embodiment, the kit include detection
Reagent R1, R2 and HCY calibration object solution.
The formula of reagent R1 are as follows: the Tris buffer of 50mM, pH8.5, Triton X-100 1g/L, mannitol 5g/L, uncle
Butylhydroquinone 1mM, disodium ethylene diamine tetraacetate 0.5mM, three (2- carbonylethyl) microcosmic salt hydrochlorate 2mM, S- adenosine first sulphur ammonia
Sour 1mM, α-ketoglutaric acid 5mM, NADH 0.4mM, NaN3 1g/L。
The formula of reagent R2 are as follows: the HEPES buffer solution of 50mM, pH9.0, Triton X-100 1g/L, mannitol 5g/L,
Adenosylhomocysteinase EC3.3.1.1 3KU/L, Hcy transmethylase 5KU/L, adenosine deaminase 2KU/L, glutamte dehydrogenase
10KU/L, NaN3 1g/L。
Concentration is respectively 2 μM, 8 μM, 15 μM, 30 μM, 60 μM of HCY calibration object solution.
Kit the preparation method is as follows:
Reagent preparation R1: weighing each component in proportion, first prepares the Tris buffer of 50mM, and stirring and dissolving adjusts solution
PH value is 8.5, sequentially adds Triton X-100, mannitol, tert-butyl hydroquinone, disodium ethylene diamine tetraacetate, three (2-
Carbonylethyl) microcosmic salt hydrochlorate, S-adenosylmethionine, α-ketoglutaric acid, NADH and NaN3, mix, after stirring and dissolving, with purifying
Water is settled to preformulation volume, with 0.45 μm of membrane filtration, after mixing, stores in 2 DEG C~8 DEG C.
Reagent preparation R2: weighing each component in proportion, first prepares the HEPES buffer solution of 50mM, and stirring and dissolving adjusts solution
PH value 9.0, sequentially adds Triton X-100, mannitol, Adenosylhomocysteinase EC3.3.1.1, Hcy transmethylase,
Adenosine deaminase, glutamte dehydrogenase and NaN3, mix, after stirring and dissolving, preformulation volume be settled to purified water, with 0.45
μm membrane filtration after mixing, is stored in 2 DEG C~8 DEG C.
It prepares calibration object solution: first preparing the 50mM PBS buffer solution of BSA containing 10g/L and 1g/L Proclin-300, with
It for preparation of reagents concentration is respectively 2 μM, 8 μM, 15 μM, 30 μM, 60 μM of HCY calibration object solution.
By the above-mentioned reagent prepared, dispensed according to 60mL/ bottles and 20mL/ bottles of reagent R2 of ratio of R1；HCY
Calibration object solution is dispensed according to 1mL/ branch, then by after packing reagent and calibration object label mounted box, as half Guang ammonia of homotype
Sour Enzymatic cycling detection kit.
The application of 2 homocysteine detection kit of embodiment
1, instrument: HITACHI 7180, OLYMPUS AU400 and TOSHIBA 40FR automatic clinical chemistry analyzer.
2, sample to be tested: the blood plasma or serum that clinical criteria method is collected, 2 DEG C~8 DEG C are 2 days storable, and -25 DEG C~-15
DEG C can be reserved for 1 month.
3, the homocysteine detection kit prepared using embodiment 1, specific detecting step are shown in Table 1.
Table 1 detects operating procedure
The concentration of calibration object is as follows: HCY concentration is 2 μm of ol/L, 8 μm of ol/L, 15 μm of ol/L, 30 μm of ol/L, 60 μm of ol/L
Calibration object solution.
4, calculated result: concentration (μm ol/L)=(Δ A of HCY in sampleSample/min-ΔABlank/min)/(ΔACalibration/min-
ΔABlank/ min) × calibration solution concentration
In formula: Δ ASample/ min refers to sample cell absorbance change rate per minute.
ΔABlank/ min refers to blank tube absorbance change rate per minute.
ΔACalibration/ min refers to that absorbance change rate per minute is managed in calibration.
5, accuracy: measurement SRM 1955, relative deviation should be no more than ± 10%.
6, sensitivity for analysis: the absolute value of the absorbance change rate (Δ A/min) of 10 μm of ol/L answers >=0.01A.
7, precision: CV≤3% in batch；CV≤5% between batch.
8, stability: 2~8 DEG C sealing stored protected from light 18 months.
The linear detection range of 3 homocysteine detection kit of embodiment is tested
The HCY sample comparison that concentration is 60 μm of ol/L is diluted to different gradient concentrations, the reagent prepared with embodiment 1
Box is averaged each concentration replication 3 times, calculates linearly dependent coefficient with theoretical value, and testing result is as shown in table 2,
Linear dependence is as shown in Figure 1.
2 HCY gradient concentration testing result of table
The corresponding linearly related equation of Fig. 1 are as follows: y=0.967x+0.7348, coefficient R2=0.9988, the results showed that
This kit linear detection range is 2~60 μm of ol/L, and Monitoring lower-cut is up to 2 μm of ol/L.
The accuracy of 4 homocysteine detection kit of embodiment is tested
The kit replication HCY international reference materials SRM 1955 prepared with embodiment 1, each concentration level repeat
Detection three times, is averaged, by formula: relative deviation=(measurement mean value-sign value)/sign value × 100% calculates opposite
Deviation, according to the accuracy for judging kit with standard substance relative deviation.
3 HCY accuracy testing result of table
| Detectable concentration | LevelⅠ | LevelⅡ | LevelⅢ |
| Repeat 1 (μm ol/L) | 4.16 | 8.75 | 17.05 |
| Repeat 2 (μm ol/L) | 4.25 | 9.13 | 16.52 |
| Repeat 3 (μm ol/L) | 3.99 | 8.98 | 16.87 |
| It measures mean value (μm ol/L) | 4.13 | 8.95 | 16.81 |
| Sign value (μm ol/L) | 3.98 | 8.85 | 17.70 |
| Relative deviation | 3.85% | 1.17% | - 5.01% |
Table 3 the result shows that, HCY reagent measurement mark 1,955 3 horizontal departures of object SRM be respectively less than 10%, illustrate this reagent
Box accuracy in detection is preferable.
The test of 5 homocysteine detection kit line withinrun precision of embodiment
The kit retest high concentration quality-control product (20 μm of ol/L) and low concentration quality-control product (10 μ prepared with embodiment 1
Mol/L sample) each 10 times, the mean value of measured value is calculated separatelyWith standard deviation (s).By formula:It calculates the coefficient of variation (CV), judges the precision of kit.
4 HCY precision testing result of table
Table 4 the result shows that, HCY reagent measurement high concentration quality-control product and the coefficient of variation CV of low concentration quality-control product are respectively less than
3%, illustrate that this kit detection precision is preferable.
The test of 6 serum sample of embodiment
The HCY reagent of kit and DIAZYME company prepared by embodiment 1 carries out sample contrastive test, the measurement knot of table 5
Fruit correlation coefficient r2It is 0.9926, correlation is good, shows that this kit can be used for clinical diagnosis.
5 HCY clinical sample comparison result of table
The constituent optimization of 7 detection reagent R1 of embodiment is tested
HCY detection reagent is prepared according to the method for embodiment 1, to constituent part (Tris buffer, Triton in reagent R1
X-100, mannitol, tert-butyl hydroquinone) content optimize, 8 group of formula (table 6) is obtained, by this 8 group of formula prepare
R1 and R2 form HCY reagent, uncap and be placed in 7180 Biochemical Analyzer of Hitachi, operated according to kit specification, detection is same
One sample, observation different formulations reagent are uncapped 1 day, 3 days, 5 days, 7 days, 10 days, 14 days steadiness in Biochemical Analyzer,
The results are shown in Table 7.Analysis data obtain optimal protective agent component.
The optimization experiment of 6 reagent R1 constituent part of table
| Component number | Tris(mM) | Triton X-100(g/L) | Mannitol (g/L) | Tert-butyl hydroquinone (mM) |
| 1 | 50 | 0.5 | 1 | 2 |
| 2 | 50 | 0.5 | 5 | 1 |
| 3 | 50 | 1 | 1 | 2 |
| 4 | 50 | 1 | 5 | 1 |
| 5 | 100 | 0.5 | 1 | 2 |
| 6 | 100 | 0.5 | 5 | 1 |
| 7 | 100 | 1 | 1 | 2 |
| 8 | 100 | 1 | 5 | 1 |
7 reagent of table is uncapped stability result
Table 7 the result shows that, the HCY reagent that different component formula is prepared is uncapped under preservation condition, and different time is uncapped reagent
It is smaller to the variation of same sample measured value compared with preparing same day reagent, it has good stability, wherein the measured value in 7 days of component 4 changes
It is relatively small, as optimization formula.
Although above the present invention is described in detail with a general description of the specific embodiments,
On the basis of the present invention, it can be modified or is improved, this will be apparent to those skilled in the art.Cause
This, these modifications or improvements, fall within the scope of the claimed invention without departing from theon the basis of the spirit of the present invention.
Claims (7)
1. a kind of stable homocysteine Enzymatic cycling detection kit, which is characterized in that the kit includes reagent
R1 and R2；
The formula of the reagent R1 are as follows: the buffer of 20-200mM, pH7.5~9.5, surfactant 0.5-5g/L, protective agent
0.5-20g/L, antioxidant 0.05-50mM, disodium ethylene diamine tetraacetate 0.2-20mM, three (2- carbonylethyl) microcosmic salt hydrochlorates
0.2-10mM, S-adenosylmethionine 0.05-5mM, α-ketoglutaric acid 1-20mM, NADH 0.1-10mM, preservative 0.5-2g/
L；
The formula of the reagent R2 are as follows: the buffer of 20-200mM, pH7.5~9.5, surfactant 0.5-20g/L, protective agent
0.5-5g/L, Adenosylhomocysteinase EC3.3.1.1 1-10KU/L, Hcy transmethylase 2-20KU/L, adenosine deaminase 1-
10KU/L, glutamte dehydrogenase 1-20KU/L, preservative 0.5-2g/L；
Wherein, it is slow to be selected from borate buffer, Triethanolamine buffer, Tris buffer, MES buffer, HEPES for the buffer
At least one of fliud flushing, PIPES buffer, CAPSO-Na buffer, PBS buffer solution.
2. detection kit according to claim 1, which is characterized in that the surfactant be selected from Tween-20,
At least one of Triton X-100, TX-10, ON-870, F68, Brij35, cetyl chloride ammonium.
3. detection kit according to claim 1, which is characterized in that the protective agent is selected from BSA, sucrose, lactose, sea
At least one of algae sugar, D-sorbite, mannitol.
4. detection kit according to claim 1, the antioxidant be selected from ascorbic acid, tert-butyl hydroquinone,
At least one of thio-2 acid, butylated hydroxy anisole, dibutyl hydroxy toluene.
5. detection kit according to claim 1, which is characterized in that the preservative is NaN3 and/or Proclin-
300。
6. detection kit according to claim 1-5, which is characterized in that the detection kit further includes
HCY calibration object solution, the concentration of the HCY calibration object solution are respectively 2 μM, 8 μM, 15 μM, 30 μM, 60 μM；For preparing
The reagent for stating HCY calibration object solution is the 50mM PBS buffer solution of BSA containing 10g/L and 1g/L Proclin-300.
7. detection kit according to claim 6, which is characterized in that the detection kit is by reagent R1, R2 and HCY
Calibration object solution composition；
The formula of the reagent R1 are as follows: the Tris buffer of 50mM, pH8.5, Triton X-100 1g/L, mannitol 5g/L, uncle
Butylhydroquinone 1mM, disodium ethylene diamine tetraacetate 0.5mM, three (2- carbonylethyl) microcosmic salt hydrochlorate 2mM, S- adenosine first sulphur ammonia
Sour 1mM, α-ketoglutaric acid 5mM, NADH0.4mM, NaN31g/L；
The formula of the reagent R2 are as follows: the HEPES buffer solution of 50mM, pH9.0, Triton X-100 1g/L, mannitol 5g/L,
Adenosylhomocysteinase EC3.3.1.1 3KU/L, Hcy transmethylase 5KU/L, adenosine deaminase 2KU/L, glutamte dehydrogenase
10KU/L, NaN31g/L；
Concentration is respectively 2 μM, 8 μM, 15 μM, 30 μM, 60 μM of HCY calibration object solution.
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| CN111304283A (en) * | 2020-03-05 | 2020-06-19 | 安徽大千生物工程有限公司 | Kit for determining HCY based on cyclic enzyme rate method and preparation and use methods thereof |
| CN113075139A (en) * | 2021-03-29 | 2021-07-06 | 迪瑞医疗科技股份有限公司 | Stable double-reagent blood ammonia determination kit |
| CN114686559A (en) * | 2020-12-31 | 2022-07-01 | 苏州博源医疗科技有限公司 | Kit for detecting homocysteine content in biological sample and preparation and use methods thereof |
| CN115655848A (en) * | 2022-12-26 | 2023-01-31 | 河北盛华尔生物医疗科技有限公司 | Stable glutamic-oxaloacetic transaminase determination kit |
| CN116165309A (en) * | 2022-06-16 | 2023-05-26 | 苏州帕诺米克生物科技有限公司 | Dilution solvent, homocysteine quality control product and application |
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| CN118130782A (en) * | 2024-05-07 | 2024-06-04 | 山东康华生物医疗科技股份有限公司 | Dry immunofluorescence quantitative detection kit for homocysteine |
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