Latex enhanced immunoturbidimetry detection reagent and application thereof

Abstract

The invention discloses a latex enhanced immunoturbidimetry detection reagent, and relates to the technical field of medical detection. Comprising the following components: an inorganic salt, a chelating agent, a coagulant, an excipient, a nonionic surfactant, an antibiotic, a preservative, an antibody stabilizer, a suspending aid, latex microspheres labeled with antibodies, high-purity liquid paraffin, amphoteric organic ionic salt, amino-terminated water-soluble hyperbranched polyamide, 1, 2-propylene glycol, a dispersant, a dispersant, a dispersant, a dispersant, a dispersant and a dispersant. The kit comprises 1, 3-bis ((trihydroxymethyl) methylamino) propane, hexahydro-1, 3-bis ((trihydroxymethyl) methylamino) propane, hexahydro-1, 3-s-triazine and a buffer solution. The latex enhanced immunoturbidimetric detection reagent is good in stability, high in repeatability, good in sensitivity and long in shelf life.

Description

Technical Field

[0001] The invention relates to the technical field of medical detection, in particular to a latex enhanced immunoturbidimetric detection reagent and application thereof. Background Art

[0002] Latex enhanced immunoturbidimetry detection reagent uses specific antibodies to coat latex particles and bind to antigens in the sample. It is a new type of immunodiagnostic reagent that achieves quantitative analysis of the sample by detecting the turbidity of the reaction solution. It has the advantages of high detection sensitivity, strong specificity, fast detection speed, simple operation, low cost, etc., and can be tested on a large scale on various biochemical analysis instruments. It is widely used in the clinical diagnosis of cardiovascular and cerebrovascular diseases, tumors, kidney diseases, liver diseases, diabetes and other diseases, and has become one of the mainstream methods for clinical quantitative detection of serum protein.

[0003] At present, the latex-enhanced immunoturbidimetric detection reagents and kits available on the market are double reagents, consisting of a reaction buffer 1 (R1) and an antibody-labeled latex-enhanced particle reagent 2 (R2). Premixing R1 and R2 together can easily cause the inactivation of the antibody in R2, aggregation of latex particles, and the inability to store the kit for a long time. Especially for the determination of whole blood samples, the currently available kits require two steps of hemolysis and determination, which is time-consuming and has poor repeatability.

[0004] In order to solve the above problems, the Chinese invention patent with the authorization announcement number CN104049085B discloses a CRP latex enhanced immune turbidimetric assay reagent, which includes latex particles, buffer, surfactant, inorganic salt, stabilizer, suspending agent, excipient and preservative labeled with CRP antibody. The invention also provides a single reagent CRP latex enhanced immune turbidimetric assay kit and its application. The invention reagent composition is simple, the operation is simple and convenient, the reaction is fast, the reagent stability is good, the sensitivity is high, the repeatability is good, the instrument is simple to use, the instrument is widely adapted, the cost is low, and it can be widely used in large, medium and small hospitals, and the evaluation of outpatient clinical rapid diagnosis of infection and infection prognosis. However, it only discloses that it is applicable to CRP projects, and there is still a lack of a general latex enhanced immune turbidimetric detection reagent applicable to multiple projects (including CRP) on the market. And the repeatability, sensitivity, stability and shelf life of the reagent detection still have room for improvement.

[0005] It can be seen that the development of a single-reagent latex-enhanced immunoturbidimetric detection reagent with good stability, high repeatability, excellent sensitivity and long shelf life and its application meet market demand, have broad market value and application prospects, and are of great significance to promoting the development of the field of latex-enhanced immunoturbidimetric detection. Summary of the invention

[0006] The purpose of the present invention is to overcome the deficiencies of the prior art and to provide a single-reagent latex-enhanced immunoturbidimetric detection reagent with good stability, high repeatability, good sensitivity and long shelf life and its application.

[0007] To achieve the above object, the technical scheme adopted by the present invention is: a latex enhanced immunoturbidimetric detection reagent, comprising the following components in percentage by weight: 0.5-2wt% of inorganic salt, 0.1-1wt% of chelating agent, 0.4-2.2wt% of coagulant, 0.9-5.2wt% of excipient, 0.5-1wt% of nonionic surfactant, 0.1-0.3wt% of antibiotic, 0.04-0.6wt% of preservative, 0.4wt% of antibody stabilizer -2.2wt%, suspending agent 0.9-5wt%, latex microspheres labeled with antibodies 0.1-0.2wt%, high-purity liquid paraffin 0.1-0.6wt%, amphoteric organic ion salt 0.1-0.3wt%, amino-terminated water-soluble hyperbranched polyamide 0.01-0.12wt%, 1,3-bis((trihydroxymethyl)methylamino)propane 0.03-0.1wt%, hydroxyethylhexahydro-s-triazine 0.01-0.06wt%, and the balance is buffer solution.

[0008] Preferably, the inorganic salt is one or a combination of sodium chloride, potassium chloride, sodium sulfate, potassium sulfate.

[0009] Preferably, the chelating agent is ethylenediaminetetraacetic acid.

[0010] Preferably, the coagulant is at least one of PEG6000 and dextran.

[0011] Preferably, the excipient is at least one of mannitol, glucose, sucrose and maltose.

[0012] Preferably, the nonionic surfactant is at least one of Triton X-100 and Tween 40.

[0013] Preferably, the antibiotic is gentamicin; the preservative is at least one of sodium azide and ProClin300; and the antibody stabilizer is at least one of bovine serum albumin and casein.

[0014] Preferably, the suspending agent is at least one of ethylene glycol and glycerol.

[0015] Preferably, the antibody-labeled latex microspheres are latex microspheres labeled with anti-target detection antibody, which can be prepared according to conventional methods in the art.

[0016] Preferably, the latex microspheres are at least one of polystyrene latex microspheres containing carboxyl groups on the surface or polystyrene latex microspheres containing amino groups on the surface.

[0017] Preferably, the latex microspheres include at least two different sizes of latex microspheres with average particle sizes of 60-150 nm and 300-450 nm.

[0018] Preferably, the surface carboxyl density of the polystyrene latex microspheres containing carboxyl groups on the surface is 0.05-0.35 mmol / g, and the surface amino density of the polystyrene latex microspheres containing amino groups on the surface is 0.1-0.2 mmol / g.

[0019] Preferably, the high-purity liquid paraffin is No. 26 white oil with a purity of ≥99.9%, provided by Jingmen Petrochemical.

[0020] Preferably, the amphoteric organic ion salt is at least one of hydroxypropane sulfonate pyridinium salt and 1-benzylpyridine-3-carboxylate.

[0021] Preferably, the amino-terminated water-soluble hyperbranched polyamide is prepared according to the method of Example 1 of the Chinese invention patent with authorization publication number CN1232567C.

[0022] Preferably, the buffer is any one of 3-[N,N-di(hydroxyethyl)amino]-2-hydroxypropanesulfonic acid-sodium hydroxide buffer (DIPSO-NaOH), 4-(2-hydroxyethyl)-1-piperazinepropanesulfonic acid-sodium hydroxide buffer (HEPPS-NaOH), 3-(N-morpholino)propanesulfonic acid-sodium hydroxide buffer (MOPS-NaOH), N-(2-hydroxyethyl)piperazine-N'-2-ethanesulfonic acid-NaOH buffer (HEPES-NaOH), tris(hydroxymethylaminomethane)-HCl buffer (Tris-HCl), phosphate buffer, glycine-NaOH buffer, and barbiturate buffer. The adjustment ability of the buffer requires a pH range of 7.0-9.0 and a concentration range of 10-100 mmol / L.

[0023] Another object of the present invention is to provide a method for preparing the latex-enhanced immunoturbidimetric detection reagent, comprising the following steps: uniformly mixing the components of the single-reagent latex-enhanced immunoturbidimetric detection reagent by weight percentage to obtain the latex-enhanced immunoturbidimetric detection reagent.

[0024] Another object of the present invention is to provide an application of the latex-enhanced immunoturbidimetric detection reagent in a single-reagent CRP latex-enhanced immunoturbidimetric assay kit.

[0025] Due to the application of the above technical solution, the present invention has the following beneficial effects:

[0026] (1) The preparation method of the latex enhanced immunoturbidimetric detection reagent disclosed in the present invention has simple process, convenient operation and control, high preparation efficiency and finished product qualification rate, is suitable for Lenovo large-scale production, and has high promotion and application value.

[0027] (2) The latex enhanced immunoturbidimetric detection reagent disclosed in the present invention is prepared by including the following components in percentage by weight: 0.5-2wt% of inorganic salt, 0.1-1wt% of chelating agent, 0.4-2.2wt% of coagulant, 0.9-5.2wt% of excipient, 0.5-1wt% of nonionic surfactant, 0.1-0.3wt% of antibiotic, 0.04-0.6wt% of preservative, 0.4-2.2wt% of antibody stabilizer. %, suspending agent 0.9-5wt%, latex microspheres labeled with antibodies 0.1-0.2wt%, high-purity liquid paraffin 0.1-0.6wt%, amphoteric organic ion salt 0.1-0.3wt%, amino-terminated water-soluble hyperbranched polyamide 0.01-0.12wt%, 1,3-bis((trihydroxymethyl)methylamino)propane 0.03-0.1wt%, hydroxyethyl hexahydro-s-triazine 0.01-0.06wt%, and the balance is buffer. Through the mutual coordination and joint action of the components, the prepared latex-enhanced immunoturbidimetric detection reagent has good stability, high repeatability, good sensitivity, and long shelf life.

[0028] (3) The latex-enhanced immunoturbidimetric detection reagent disclosed in the present invention adopts a single reagent, which is simpler to operate than the traditional double reagent and can effectively avoid antibody inactivation and latex particle aggregation; the latex microspheres at least include two different sizes of latex microspheres with average particle sizes of 60-150nm and 300-450nm, which increases the turbidity during antibody reaction, thereby increasing the sensitivity of the test reaction, and at the same time ensuring the linearity of the reagent. The addition of amino-terminated water-soluble hyperbranched polyamide, 1,3-bis((trihydroxymethyl)methylamino)propane, hydroxyethylhexahydro-s-triazine, and amphoteric organic ion salts, and the interaction between other raw material components, can bind to antibodies through weak interactions such as hydrogen bonds, on the one hand, inhibiting the latex agglomeration phenomenon caused by the binding of other non-specific proteins or small molecules to antibody-sensitized latex, and on the other hand, it does not affect the specific binding of antibodies to antigens. Thereby improving the stability and detection accuracy of the reagent and improving the detection range.

[0029] (4) The latex-enhanced immunoturbidimetric detection reagent disclosed in the present invention can prevent the volatilization of the liquid by adding high-purity liquid paraffin, and cooperates with the amphoteric organic ion salt to effectively improve the stability of the reagent. DETAILED DESCRIPTION

[0030] The following description is used to disclose the present invention so that those skilled in the art can implement the present invention. The preferred embodiments described below are only examples, and those skilled in the art may think of other obvious variations.

[0031] Example 1

[0032] A latex enhanced immunoturbidimetric detection reagent is prepared by comprising the following components in percentage by weight: 0.5wt% of inorganic salt, 0.1wt% of chelating agent, 0.4wt% of coagulant, 0.9wt% of excipient, 0.5wt% of nonionic surfactant, 0.1wt% of antibiotic, 0.04wt% of preservative, 0.4wt% of antibody stabilizer, 0.9wt% of suspending agent, 0.1wt% of latex microspheres labeled with antibodies, 0.1wt% of high-purity liquid paraffin, 0.1wt% of amphoteric organic ion salt, 0.01wt% of amino-terminated water-soluble hyperbranched polyamide, 0.03wt% of 1,3-bis((trihydroxymethyl)methylamino)propane, 0.01wt% of hydroxyethylhexahydro-s-triazine, and the balance is buffer solution.

[0033] The inorganic salt is sodium chloride; the chelating agent is ethylenediaminetetraacetic acid; the coagulant is PEG6000; the excipient is mannitol; the nonionic surfactant is Triton X-100; the antibiotic is gentamicin; the preservative is sodium azide; the antibody stabilizer is bovine serum albumin; the suspending agent is ethylene glycol; the antibody-labeled latex microspheres are latex microspheres labeled with anti-target detection antibody, prepared according to conventional methods in the art (refer to the method in CN104049085) ; The latex microspheres are polystyrene latex microspheres containing carboxyl groups on the surface and polystyrene latex microspheres containing amino groups on the surface, which are mixed in a mass ratio of 1:1; the average particle size of the polystyrene latex microspheres containing carboxyl groups on the surface is 60nm, and the average particle size of the polystyrene latex microspheres containing amino groups on the surface is 450nm; the surface carboxyl density of the polystyrene latex microspheres containing carboxyl groups on the surface is 0.05mmol / g, and the surface amino density of the polystyrene latex microspheres containing amino groups on the surface is 0.1mmol / g.

[0034] The high-purity liquid paraffin is No. 26 white oil with a purity of ≥99.9%, provided by Jingmen Petrochemical; the amphoteric organic ion salt is hydroxypropane sulfonic acid pyridinium salt; the amino-terminated water-soluble hyperbranched polyamide is prepared according to the method of Example 1 of the Chinese invention patent with authorization announcement number CN1232567C.

[0035] The buffer is 3-[N,N-di(hydroxyethyl)amino]-2-hydroxypropanesulfonic acid-sodium hydroxide buffer (DIPSO-NaOH), and the adjustment capacity of the buffer requires a pH range of 7.0-9.0 and a concentration range of 10 mmol / L.

[0036] A preparation method of the latex enhanced immunoturbidimetric detection reagent comprises the following steps: uniformly mixing the components of the single-reagent latex enhanced immunoturbidimetric detection reagent according to weight percentage to obtain the latex enhanced immunoturbidimetric detection reagent.

[0037] Example 2

[0038] A latex enhanced immunoturbidimetric detection reagent is prepared by comprising the following components in percentage by weight: 0.9wt% of inorganic salt, 0.3wt% of chelating agent, 1.2wt% of coagulant, 2.2wt% of excipient, 0.6wt% of nonionic surfactant, 0.15wt% of antibiotic, 0.15wt% of preservative, 0.9wt% of antibody stabilizer, 2wt% of suspending agent, 0.13wt% of latex microspheres labeled with antibodies, 0.2wt% of high-purity liquid paraffin, 0.15wt% of amphoteric organic ion salt, 0.05wt% of amino-terminated water-soluble hyperbranched polyamide, 0.06wt% of 1,3-bis((trihydroxymethyl)methylamino)propane, 0.02wt% of hydroxyethylhexahydro-s-triazine, and the balance is buffer solution.

[0039] The inorganic salt is potassium chloride; the chelating agent is ethylenediaminetetraacetic acid; the coagulant is dextran; the excipient is glucose; the nonionic surfactant is Tween 40; the antibiotic is gentamicin; the preservative is ProClin300; the antibody stabilizer is casein; the suspending agent is ethylene glycol; the antibody-labeled latex microspheres are latex microspheres labeled with anti-target detection antibody, which are prepared according to conventional methods in the art (refer to the method in CN104049085); The latex microspheres are polystyrene latex microspheres containing carboxyl groups on the surface and polystyrene latex microspheres containing amino groups on the surface, which are mixed in a mass ratio of 2:3; the average particle size of the polystyrene latex microspheres containing carboxyl groups on the surface is 100 nm, and the average particle size of the polystyrene latex microspheres containing amino groups on the surface is 420 nm; the surface carboxyl density of the polystyrene latex microspheres containing carboxyl groups on the surface is 0.15 mmol / g, and the surface amino density of the polystyrene latex microspheres containing amino groups on the surface is 0.13 mmol / g.

[0040] The high-purity liquid paraffin is No. 26 white oil with a purity of ≥99.9%, provided by Jingmen Petrochemical; the amphoteric organic ion salt is 1-benzylpyridine-3-carboxylate; the amino-terminated water-soluble hyperbranched polyamide is prepared according to the method of Example 1 of the Chinese invention patent with authorization announcement number CN1232567C; the buffer solution is 4-(2-hydroxyethyl)-1-piperazinepropanesulfonic acid-sodium hydroxide buffer solution (HEPPS-NaOH), and the adjustment ability of the buffer solution requires a pH range of 7.0-9.0 and a concentration range of 30mmol / L.

[0041] A preparation method of the latex enhanced immunoturbidimetric detection reagent comprises the following steps: uniformly mixing the components of the single-reagent latex enhanced immunoturbidimetric detection reagent according to weight percentage to obtain the latex enhanced immunoturbidimetric detection reagent.

[0042] Example 3

[0043] A latex enhanced immunoturbidimetric detection reagent is prepared by comprising the following components in percentage by weight: 1.3 wt% of inorganic salt, 0.6 wt% of chelating agent, 1.4 wt% of coagulant, 3.2 wt% of excipient, 0.8 wt% of nonionic surfactant, 0.2 wt% of antibiotic, 0.35 wt% of preservative, 1.4 wt% of antibody stabilizer, 3.5 wt% of suspending agent, 0.15 wt% of latex microspheres labeled with antibodies, 0.4 wt% of high-purity liquid paraffin, 0.2 wt% of amphoteric organic ion salt, 0.08 wt% of amino-terminated water-soluble hyperbranched polyamide, 0.07 wt% of 1,3-bis((trihydroxymethyl)methylamino)propane, 0.035 wt% of hydroxyethylhexahydro-s-triazine, and the balance is buffer solution.

[0044] The inorganic salt is sodium chloride, potassium chloride, sodium sulfate, and potassium sulfate mixed in a mass ratio of 1:2:1:2; the chelating agent is ethylenediaminetetraacetic acid; the coagulant is PEG6000 and dextran mixed in a mass ratio of 1:3; the excipient is mannitol, glucose, sucrose, and maltose mixed in a mass ratio of 1:2:2:3; the non-ionic surfactant is Triton X-100 and Tween 40 mixed in a mass ratio of 3:5; the antibiotic is gentamicin; the preservative is sodium azide and ProClin300 mixed in a mass ratio of 2:3; the antibody stabilizer is bovine serum albumin and casein mixed in a mass ratio of 3:1; the suspending agent is ethylene glycol and glycerol mixed in a mass ratio of 2:3.

[0045] The antibody-labeled latex microspheres are latex microspheres labeled with anti-target detection antibody, which are prepared according to conventional methods in the art (refer to the method in CN104049085); the latex microspheres are polystyrene latex microspheres containing carboxyl groups on the surface and polystyrene latex microspheres containing amino groups on the surface, which are mixed in a mass ratio of 1:3; the average particle size of the polystyrene latex microspheres containing carboxyl groups on the surface is 130nm; the average particle size of the polystyrene latex microspheres containing amino groups on the surface is 350nm; the surface carboxyl density of the polystyrene latex microspheres containing carboxyl groups on the surface is 0.3mmol / g, and the surface amino density of the polystyrene latex microspheres containing amino groups on the surface is 0.18mmol / g.

[0046] The high-purity liquid paraffin is No. 26 white oil with a purity of ≥99.9%, provided by Jingmen Petrochemical; the amphoteric organic ion salt is a mixture of hydroxypropane sulfonic acid pyridinium salt and 1-benzylpyridine-3-carboxylate in a mass ratio of 1:1; the amino-terminated water-soluble hyperbranched polyamide is prepared according to the method of Example 1 of the Chinese invention patent with authorization announcement number CN1232567C; the buffer solution is 3-(N-morpholinyl)propanesulfonic acid-sodium hydroxide buffer solution (MOPS-NaOH), and the adjustment ability of the buffer solution requires a pH range of 7.0-9.0 and a concentration range of 60mmol / L.

[0047] A preparation method of the latex enhanced immunoturbidimetric detection reagent comprises the following steps: uniformly mixing the components of the single-reagent latex enhanced immunoturbidimetric detection reagent according to weight percentage to obtain the latex enhanced immunoturbidimetric detection reagent.

[0048] Example 4

[0049] A latex enhanced immunoturbidimetric detection reagent is prepared by comprising the following components in percentage by weight: 1.8 wt% of inorganic salt, 0.9 wt% of chelating agent, 2 wt% of coagulant, 5 wt% of excipient, 0.9 wt% of nonionic surfactant, 0.25 wt% of antibiotic, 0.5 wt% of preservative, 2 wt% of antibody stabilizer, 4.5 wt% of suspending agent, 0.18 wt% of latex microspheres labeled with antibodies, 0.5 wt% of high-purity liquid paraffin, 0.25 wt% of amphoteric organic ion salt, 0.1 wt% of amino-terminated water-soluble hyperbranched polyamide, 0.09 wt% of 1,3-bis((trihydroxymethyl)methylamino)propane, 0.05 wt% of hydroxyethylhexahydro-s-triazine, and the balance is buffer solution.

[0050] The inorganic salt is potassium sulfate; the chelating agent is ethylenediaminetetraacetic acid; the coagulant is PEG6000; the excipient is maltose; the nonionic surfactant is Triton X-100; the antibiotic is gentamicin; the preservative is sodium azide; the antibody stabilizer is bovine serum albumin; the suspending agent is glycerol; the antibody-labeled latex microspheres are latex microspheres labeled with anti-target detection antibody, which are prepared according to conventional methods in the art (refer to the method in CN104049085).

[0051] The latex microspheres are formed by mixing polystyrene latex microspheres containing carboxyl groups on the surface and polystyrene latex microspheres containing amino groups on the surface in a mass ratio of 2:1; the average particle size of the polystyrene latex microspheres containing carboxyl groups on the surface is 140nm; the average particle size of the polystyrene latex microspheres containing amino groups on the surface is 330nm; the surface carboxyl density of the polystyrene latex microspheres containing carboxyl groups on the surface is 0.33mmol / g, and the surface amino density of the polystyrene latex microspheres containing amino groups on the surface is 0.18mmol / g.

[0052] The high-purity liquid paraffin is No. 26 white oil with a purity of ≥99.9%, provided by Jingmen Petrochemical; the amphoteric organic ion salt is hydroxypropane sulfonic acid pyridinium salt; the amino-terminated water-soluble hyperbranched polyamide is prepared according to the method of Example 1 of the Chinese invention patent with authorization announcement number CN1232567C; the buffer is N-(2-hydroxyethyl)piperazine-N'-2-ethane sulfonic acid-NaOH buffer (HEPES-NaOH), and the adjustment ability of the buffer requires a pH range of 7.0-9.0 and a concentration range of 90mmol / L.

[0053] A preparation method of the latex enhanced immunoturbidimetric detection reagent comprises the following steps: uniformly mixing the components of the single-reagent latex enhanced immunoturbidimetric detection reagent according to weight percentage to obtain the latex enhanced immunoturbidimetric detection reagent.

[0054] Example 5

[0055] A latex enhanced immunoturbidimetric detection reagent is prepared by comprising the following components in percentage by weight: 2wt% of inorganic salt, 1wt% of chelating agent, 2.2wt% of coagulant, 5.2wt% of excipient, 1wt% of nonionic surfactant, 0.3wt% of antibiotic, 0.6wt% of preservative, 2.2wt% of antibody stabilizer, 5wt% of suspending agent, 0.2wt% of latex microspheres labeled with antibodies, 0.6wt% of high-purity liquid paraffin, 0.3wt% of amphoteric organic ion salt, 0.12wt% of amino-terminated water-soluble hyperbranched polyamide, 0.1wt% of 1,3-bis((trihydroxymethyl)methylamino)propane, 0.06wt% of hydroxyethylhexahydro-s-triazine, and the balance is buffer solution.

[0056] The inorganic salt is sodium sulfate; the chelating agent is ethylenediaminetetraacetic acid; the coagulant is PEG6000; the excipient is sucrose; the nonionic surfactant is Triton X-100; the antibiotic is gentamicin; the preservative is sodium azide; the antibody stabilizer is casein; the suspending agent is glycerol; the antibody-labeled latex microspheres are latex microspheres labeled with anti-target detection antibody, which are prepared according to conventional methods in the art (refer to the method in CN104049085); The latex microspheres are polystyrene latex microspheres containing carboxyl groups on the surface and polystyrene latex microspheres containing amino groups on the surface, which are mixed in a mass ratio of 1:2; the average particle size of the polystyrene latex microspheres containing carboxyl groups on the surface is 150nm; the average particle size of the polystyrene latex microspheres containing amino groups on the surface is 300nm; the surface carboxyl density of the polystyrene latex microspheres containing carboxyl groups on the surface is 0.35mmol / g, and the surface amino density of the polystyrene latex microspheres containing amino groups on the surface is 0.2mmol / g.

[0057] The high-purity liquid paraffin is No. 26 white oil with a purity of ≥99.9%, provided by Jingmen Petrochemical; the amphoteric organic ion salt is hydroxypropane sulfonic acid pyridinium salt; the amino-terminated water-soluble hyperbranched polyamide is prepared according to the method of Example 1 of the Chinese invention patent with authorization announcement number CN1232567C; the buffer solution is trishydroxymethylaminomethane-HCl buffer solution (Tris-HCl), and the adjustment ability of the buffer solution requires a pH range between 7.0-9.0 and a concentration range of 100mmol / L.

[0058] A preparation method of the latex enhanced immunoturbidimetric detection reagent comprises the following steps: uniformly mixing the components of the single-reagent latex enhanced immunoturbidimetric detection reagent according to weight percentage to obtain the latex enhanced immunoturbidimetric detection reagent.

[0059] Comparative Example 1

[0060] A latex-enhanced immunoturbidimetric detection reagent and a preparation method thereof are substantially the same as those in Example 1, except that high-purity liquid paraffin and amphoteric organic ion salt are not added.

[0061] Comparative Example 2

[0062] A latex-enhanced immunoturbidimetric detection reagent and a preparation method thereof are substantially the same as those of Example 1, except that amino-terminated water-soluble hyperbranched polyamide, 1,3-bis((trihydroxymethyl)methylamino)propane and hydroxyethylhexahydro-s-triazine are not added.

[0063] In order to further illustrate the beneficial technical effects of the latex-enhanced immunoturbidimetric detection reagents involved in the embodiments of the present invention, the latex-enhanced immunoturbidimetric detection reagents involved in Examples 1-5 and Comparative Examples 1-2 were tested for relevant performances, and the testing methods were as follows: the reagents prepared in the above examples were tested using a Beckman AU480 fully automatic biochemical analyzer, the test wavelength was 600nm, and a CRP calibrator (a commercially available CRP pure product was dissolved in 50mM Tris-HCl, pH 7.5, 0.85% NaCl, 0.5% BSA, 0.1% sodium azide, CRP concentrations of 0mg / L, 5mg / L, 15mg / L, 50mg / L, 100mg / L, 150mg / L) 4uL, then add 300uL of the reagent prepared in each case, keep constant temperature at 37℃ for 3min, read the absorbance A1 after 20s, read the absorbance A2 after incubation at 37℃ for 2 minutes and 30 seconds, then the reaction absorbance ΔA=A2-A1; first use the standard to perform multi-point calibration, and use the spline function to calculate to obtain the calibration curve. The sample can be compared with the standard curve through its absorbance change to obtain the PCT concentration in the sample. The above examples were verified for analytical sensitivity, accuracy, precision and stability.

[0064] Table 1

[0065] project Detection range Accuracy (recovery rate) Precision (CV) stability unit ng / mL ％ ％ Month Example 1 0.01-27.5 99.73 0.97 24 Example 2 0.01-27.7 99.79 0.92 24 Example 3 0.01-27.8 99.83 0.90 24 Example 4 0.01-27.8 99.88 0.86 24 Example 5 0.01-28.0 99.91 0.83 24 Comparative Example 1 0.2-28.2 98.67 1.32 18 Comparative Example 2 0.2-28.0 98.45 1.38 18

[0066] As can be seen from Table 1, the product of the embodiment of the present invention has better sensitivity, accuracy, precision and stability than the comparative example, and the addition of high-purity liquid paraffin, amphoteric organic ion salt, amino-terminated water-soluble hyperbranched polyamide, 1,3-bis((trihydroxymethyl)methylamino)propane and hydroxyethylhexahydro-s-triazine is beneficial to improving the above properties.

[0067] The above embodiments are only for illustrating the technical concept and features of the present invention, and their purpose is to enable people familiar with this technology to understand the contents of the present invention and implement them accordingly. They cannot be used to limit the protection scope of the present invention. All equivalent changes or modifications made according to the spirit of the present invention should be included in the protection scope of the present invention.

Claims

1. A latex enhanced immunoturbidimetric detection reagent, characterized in that: The invention comprises the following components in percentage by weight: 0.5-2wt% of inorganic salt, 0.1-1wt% of chelating agent, 0.4-2.2wt% of coagulant, 0.9-5.2wt% of excipient, 0.5-1wt% of nonionic surfactant, 0.1-0.3wt% of antibiotic, 0.04-0.6wt% of preservative, 0.4-2.2wt% of antibody stabilizer, 0.9-5wt% of suspending agent, 0.1-0.2wt% of latex microspheres labeled with antibodies, 0.1-0.6wt% of high-purity liquid paraffin, 0.1-0.3wt% of amphoteric organic ion salt, 0.01-0.12wt% of amino-terminated water-soluble hyperbranched polyamide, 0.03-0.1wt% of 1,3-bis((trihydroxymethyl)methylamino)propane, 0.01-0.06wt% of hydroxyethyl hexahydro-s-triazine, and the balance is buffer solution.

2. The latex-enhanced immunoturbidimetric detection reagent according to claim 1, characterized in that: The inorganic salt is one or a combination of sodium chloride, potassium chloride, sodium sulfate, potassium sulfate; and the chelating agent is ethylenediaminetetraacetic acid.

3. The latex-enhanced immunoturbidimetric detection reagent according to claim 1, characterized in that: The coagulant is at least one of PEG6000 and dextran; the excipient is at least one of mannitol, glucose, sucrose and maltose.

4. The latex-enhanced immunoturbidimetric detection reagent according to claim 1, characterized in that: The nonionic surfactant is at least one of Triton X-100 and Tween 40; and the antibiotic is gentamicin.

5. The latex-enhanced immunoturbidimetric detection reagent according to claim 1, characterized in that: The preservative is at least one of sodium azide and ProClin300; the antibody stabilizer is at least one of bovine serum albumin and casein; and the suspending agent is at least one of ethylene glycol and glycerol.

6. The latex-enhanced immunoturbidimetric detection reagent according to claim 1, characterized in that: The antibody-labeled latex microspheres are latex microspheres labeled with anti-target detection antibody; the latex microspheres are at least one of polystyrene latex microspheres containing carboxyl groups on the surface or polystyrene latex microspheres containing amino groups on the surface; the latex microspheres include at least two different sizes of latex microspheres with average particle sizes of 60-150nm and 300-450nm; the surface carboxyl density of the polystyrene latex microspheres containing carboxyl groups on the surface is 0.05-0.35mmol / g, and the surface amino density of the polystyrene latex microspheres containing amino groups on the surface is 0.1-0.2mmol / g.

7. The latex-enhanced immunoturbidimetric detection reagent according to claim 1, characterized in that: The high-purity liquid paraffin is No. 26 white oil with a purity of ≥99.9%; the amphoteric organic ion salt is at least one of hydroxypropane sulfonic acid pyridinium salt and 1-benzylpyridine-3-carboxylate.

8. The latex-enhanced immunoturbidimetric detection reagent according to claim 1, characterized in that: The buffer is any one of 3-[N,N-di(hydroxyethyl)amino]-2-hydroxypropanesulfonic acid-sodium hydroxide buffer (DIPSO-NaOH), 4-(2-hydroxyethyl)-1-piperazinepropanesulfonic acid-sodium hydroxide buffer (HEPPS-NaOH), 3-(N-morpholino)propanesulfonic acid-sodium hydroxide buffer (MOPS-NaOH), N-(2-hydroxyethyl)piperazine-N'-2-ethanesulfonic acid-NaOH buffer (HEPES-NaOH), tris(hydroxymethylaminomethane)-HCl buffer (Tris-HCl), phosphate buffer, glycine-NaOH buffer, and barbiturate buffer. The buffer has an adjustment capability of a pH range of 7.0-9.0 and a concentration range of 10-100 mmol / L.

9. A method for preparing the latex enhanced immunoturbidimetric detection reagent according to any one of claims 1 to 8, characterized in that: The method comprises the following steps: uniformly mixing the components of a single-reagent latex-enhanced immunoturbidimetric detection reagent according to weight percentage to obtain the latex-enhanced immunoturbidimetric detection reagent.

10. Use of the latex-enhanced immunoturbidimetric detection reagent according to any one of claims 1 to 8 in a single-reagent CRP latex-enhanced immunoturbidimetric assay kit.