CN107860930A

CN107860930A - The immunoturbidimetry detection reagent and method of a kind of cardic fatty acid binding protein

Info

Publication number: CN107860930A
Application number: CN201711102671.9A
Authority: CN
Inventors: 刘向晖; 陈胜胜; 陶艳茹
Original assignee: SUZHOU KANGHESHUN MEDICAL TECHNOLOGY Co Ltd
Current assignee: SUZHOU KANGHESHUN MEDICAL TECHNOLOGY Co Ltd
Priority date: 2017-11-10
Filing date: 2017-11-10
Publication date: 2018-03-30

Abstract

The invention discloses a kind of cardic fatty acid binding protein（H‑FABP,Heart‑type fatty acid binding protein）Latex enhancing immune than turbid detection reagent.The reagent is single reagent, and reagent main component is the latex particle of H FABP antibody labelings, while also includes cushioning liquid, surfactant, salt, stabilizer, suspending agent and preservative.The invention also discloses utilize transmission or scattering turbidimetry principle detection blood sample centre type fatty acid binding protein using the reagent（H‑FABP）Concentration method.The present invention uses single agents, simple to operate, without mixing, high sensitivity, the range of linearity is wide, being capable of the various samples such as direct measurement whole blood, serum and blood plasma, it can be widely applied to various transmissions or scattering analysis instrument, including common Biochemical Analyzer, specific protein analyzer etc..

Description

The immunoturbidimetry detection reagent and method of a kind of cardic fatty acid binding protein

Technical field

The invention belongs to medical immunology diagnostic field, and in particular to a kind of cardic fatty acid binding protein（H-FABP）Glue The preparation and its application of breast enhancing immunoturbidimetry detection reagent.

Background technology

Cardic fatty acid binding protein (H-FABP) is a kind of new small cytoplasmic protein being rich in heart.Myocardial ischemia Damage occur after, H-FABP can after episode 1-3 hours be found in blood, 6-8 hours reach peak value and Blood plasma level recovers normal within 24-30 hours.Acute coronary artery syndrome (ACS) be acute events important in coronary heart disease it One, it is divided into acute myocardial infarction AMI (AMI), unstable angina (UA) and sudden cardiac death (SCD), its incidence of disease and case fatality rate It is higher.But if high-risk patient can be distinguished early after morbidity and carry out reperfusion as treatment, case fatality rate and prognosis have substantially Improvement.Wherein, cardiac marker plays the part of important role during ACS diagnosis and prognosis.Conventional cardiac marker has Troponin (cTn, including cTnI and cTnT), myoglobins (MYO) and creatine kinase isozyme (CK-MB) etc..Due to ACS It is anxious with morbidity, endanger the characteristics of big, therefore the high early sign thing of specificity is selected, diagnose ahead of time, treatment is for ACS in time Patient is significant.H-FABP (H-FABP), therefore can be myocardium thin because of its molecular weight (14.9KDa) (1.5h) quick release quickly (24h) can be recovered to normal level, for ACS again into blood through renal metabolism after cellular damage Diagnosis there is jump, while possess the characteristics of high specific, high sensitivity, high coincidence rate to myocardial damage；Grown in ACS In phase prognosis, H-FABP concentration can effectively identify the high-risk patient of the adverse events such as AMI, heart failure and unstable angina；Together When H-FABP and troponin joint-detection can improve diagnostic sensitivity, diagnostic value is had more to ACS.

H-FABP common detection method has colloidal gold method（People's H-FABP colloidal gold tests and preparation method thereof； CN201410163289）, immunochromatographic method（A kind of fluorescence immune chromatography reagent for quantitatively detecting hFABP Box；CN201110453809.6）, latex enhancing immune turbidimetry（Particles Enhanced Turbidity Immuno- Assay - PETIA）.Most advanced with latex enhancing immune turbidimetry in these methods, sensitivity is higher, and the range of linearity is wider, Also relative ease is operated, is widely adopted in clinic.Latex enhancing immune turbidimetry（PETIA）Using absorption in diameter tens The cardic fatty acid binding protein in H-FABP antibody capture test samples on to hundreds of nanometers of present latex particulates（H-FABP）, So as to form the crosslinking between particulate, the nephelometric turbidity unit or transmission absorbance of solution are changed（That is turbidity）.By measuring solution Nephelometric turbidity unit or transmit absorbance change, test sample centre type aliphatic acid can be quantitatively calculated according to calibration curve Protein-bonded concentration.The method of testing can both be used on general large-scale automatic biochemistry analyzer, can also be specific The small-sized POCT such as protein analyzer（Immediately detection）Used in equipment, there is very strong applicability, China can be met well From large-scale Grade A hospital to the use demand of community's basic hospital.

Currently on the market existing H-FABP latex enhancing immunes than turbid reagent with the cardioid aliphatic acid combination egg of RANDOX companies White detection kit (immunoturbidimetry)（State's food medicine prison tool (enters) word 2014 the 2402142nd）To represent, using dilution The double reagent form of liquid and latex solution.In test, test sample first must dilute in dilution, then enter with latex solution Row hybrid reaction, then tested in transmission or scatterometer.The operation of dilution and mixing is in automatic clinical chemistry analyzer（Front three Hospital's common instrument platform）On can be automatically brought into operation by machine, but especially POCT classes are just on small semiautomatic platform Take formula platform（Hospital below three-level and counties and townships' one-level medical services unit are commonly used）The test of upper progress then needs operating personnel Implement manually, bring extra operating procedure, have also been introduced more operating errors, have impact on the precision of test, it is unfavorable In the detection small-middle hospital promotion and implementation.H-FABP latex enhancing immunes on existing market than turbid reagent another Shortcoming is can only to measure the H-FABP contents in serum or plasma sample, must carry out extracting the preceding place of serum to whole blood sample Science and engineering is made, time and effort consuming, poor repeatability, very inconvenient in the actual use of clinic.

The content of the invention

It is an object of the invention to provide it is a kind of it is simple to operate, whole blood, blood plasma can be tested without mix reagent while directly With the single reagent cardic fatty acid binding protein of serum sample（H-FABP）Latex enhancing immune than turbid detection reagent.The reagent Both it can be tested, can also be surveyed in the small-sized POCT equipment such as special protein instrument on large-scale automatic biochemistry analyzer Examination.

To achieve the above object, a kind of technical scheme provided by the invention is a kind of cardioid aliphatic acid combination egg based on single reagent In vain（H-FABP）Latex enhancing immune is than turbid detection kit.The kit includes cardic fatty acid binding protein（H-FABP）Inspection Test agent, Heartfattyacidbindingprotein control product and cardic fatty acid binding protein standard items.

Mentioned in the introduction of background technology, the existing H-FABP latex enhancing immunes detection reagent of in the market is using double examinations The test format of agent（The kit of latex enhancing immune turbidimetry for Determination serum or urine H-FABP in rat； CN201110454515.5）.Wherein dilution is commonly referred to as R1, and latex solution is commonly referred to as R2.Pure antibody or antigen The latex solution of coated microballoon is thermodynamic unstable system, if without special processing, latex can spontaneously aggegation.Cause This, usually, the latex solution in detection reagent（R2）Need to ensure the stabilization of latex solution by optimizing the composition of solution, Prevent latex that spontaneous agglutination occurs in storage.Common optimization means include from different buffer systems, change microballoon table Surface charge, change the means such as solution ion strength, addition stabilizer（J.A. Molina Bolivar et al,Journal of Macromolecular Science Part C - Polymer Reviews,2005, 45:59-98）.But work as glue When milk solution is used to test, the latex agglutination as caused by the specific binding of antigen-antibody should not just be prevented from, and this is required Dilution（R1）The formation of latex agglutination can be beneficial to a certain extent.So general dilution（R1）With latex storing liquid （R2）Made a big difference on formula.Directly R1 and R2 is mixed, it is unstable to generally result in latex.This is also market gluing Newborn reagent typically all uses the main reason for double reagent form.

In the present invention, in addition to the method for the stabilized latex reagent of routine, we also introduce surfactant to optimize Agent prescription stabilized latex solution.Surfactant can provide steric hindrance, and the surfactant of ionic can also provide Extra electrostatic repulsion, so as to prevent the cohesion of latex, the stabilized latex reagent in storage.Meanwhile by optimizing agent prescription, When the sample contact emulsion reagent containing H-FABP, suitable surfactant can promote H-FABP antigens and H-FABP to resist The interaction of body, advantageously form specific latex agglutination.So we are just prepared for a kind of H- of single reagent form FABP emulsion reagents.In storage, the reagent, which can be stablized, to be deposited, and when contacting sample, aggegation can occur rapidly for the reagent Reaction, it is no longer necessary to mix other dilutions to promote agglutinating reaction.And preferable kinds of surfactants and concentration can With haemocyte in effective lysed blood, so that the detection reagent in the present invention can possess with the sample of direct measurement whole blood The ability of whole blood, serum and blood plasma is detected simultaneously.

Cardic fatty acid binding protein of the present invention（H-FABP）The latex that detection reagent includes H-FABP antibody labelings is micro- Ball, cushioning liquid, surfactant, salt, stabilizer, suspending agent and preservative.

Described H-FABP antibody is H-FABP more anti-either a pair or several H-FABP monoclonal antibodies to pairing.

A diameter of 50nm-400nm of described latex microsphere;Its surface modification group be-COOH ,-NH2 ,-SH ,- One kind or physisorption type latex microsphere in OH ,-CHO.

Preferably, the latex microsphere is polystyrene latex microspheres, its a diameter of 150-350nm, and surface modification base Group is-COOH.

Described cushioning liquid is glycine buffer, HEPES buffer solution, MOPSO buffer solutions, Tris buffer solutions, PBS delay One kind in fliud flushing, MES buffer solutions.

Preferably, described cushioning liquid is MES cushioning liquid, pH adjustable ranges between 5.5-8.0, concentration 10- 150mmol/L。

Described surfactant is NP40, triton x-100, polysorbas20, polysorbate40, Tween 80, SDS, span 40, department One kind, two or more combination in disk 60.Under the combination of optimization and concentration, surfactant has haemolysis simultaneously With the double effectses for maintaining stable reagent.

Preferably, the surfactant is triton x-100, NP40, one kind in polysorbas20, concentration 0.1%-1%.

Described salt be sodium chloride, potassium chloride, calcium chloride, sodium sulphate, one kind in potassium sulfate, two or more Combination.

Preferably, described salt is 0.5%-3.5% sodium chloride.

The stabilizer is one kind in bovine serum albumin(BSA), gelatin or casein.

Preferably, described stabilizer is bovine serum albumin, concentration 0.2%-2%

The suspending agent be lactose, sucrose, glucose, maltose, trehalose, one kind in mannitol, two or more Combination.

Preferably, described suspending agent is trehalose, concentration 0.3%-3%.

Described preservative is one kind in sodium azide, ProClin-300, thimerosal.

Preferably, described preservative is ProClin-300, concentration 0.05-0.2%.

The quality-control product of cardic fatty acid binding protein of the present invention is two sets of H-FABP quality-control products of high level and low value, is used In checking reagent performance.

The standard items of cardic fatty acid binding protein of the present invention are a series of H- of low value concentration to high level concentration FABP standard items, concentration range is from 0 to 120ng/mL, the calibration curve required for establishing test.

When detecting sample, the H-FABP antibody in reagent provided by the invention on latex particle is sent out with the H-FABP antigens in sample Raw antibody antigen reaction, so that latex gradually condenses, changes the turbidity of test fluid.The change of turbidity can pass through transmission Instrument or scatterometer determine.The absorbance or scattered light intensity of transmission are with the H-FABP concentration in sample in finite concentration model Enclose interior linear.The calibration curve established by reference to H-FABP standard items, quantitatively can be calculated in sample H-FABP concentration.

Single reagent H-FABP latex enhancing immunes turbidimetric assay kit provided by the invention, be adapted to various transilluminators or Scatterometer.

Beneficial effects of the present invention include：

1. the composition of detection reagent is simple, only single agents, it is not necessary to as the available reagent of market test when carry out R1, R2 married operation, simplifies operation sequence.Especially when using semi-automatic POCT equipment, pole is brought for operating personnel Big facility, while also preferably ensured the accuracy of test.

2. reagent has the function of haemolysis and detection simultaneously, whole blood sample can be directly tested, it is not necessary to carry out serum and carry The work taken, operation is simplified, substantially reduce the whole testing time including preparation of samples.Generally, serum Extraction needs the time of 30 to 120 minutes or so.

3. the reagent reacting is quick, the reaction time only needs 150 to 180 seconds.Compared to existing 10 to 12 points of emulsion reagent The detection time of clock, speed are improved close to 4 times, so as to greatly improve detection efficiency, are particularly advantageous to the quick detection of outpatient service.

4. the reagent range of linearity is broad, dilution sample of the direct measurement at concentrations up to 120ng/mL can not had to.

5. reagent applicability is good, can be used in transmission or scatterometer.Both can be in fully-automatic large-scale biochemical instruments Use, can also be used in small-sized POCT equipment.Can meet simultaneously large hospital clinical laboratory, small hospital's outpatient service and The detection of emergency treatment section office needs.

Brief description of the drawings

Calibration curve of Fig. 1 embodiments 1 in biochemical instruments

Fig. 22 calibration curves in POCT equipment of embodiment

The range of linearity of Fig. 3 embodiments 1

The range of linearity of Fig. 4 embodiments 2

The correlation of Fig. 5 embodiments 2

Embodiment

With reference to specific embodiment, the present invention is further elaborated.Following embodiments is to further illustrate Some currently preferred embodiments of the present invention, and not all embodiments.Those skilled in the art are before no progress creative work The other embodiment based on the present invention made is put, belongs to the scope of the present invention.

Embodiment 1

1）The preparation of emulsion reagent

By particle diameter for 333nm surface carboxyl groups microballoon in MES（50mM, pH=6.5）1% is diluted in cushioning liquid, at room temperature Stirring.EDC solid powders are put into, it is 30 to keep EDC and carboxyl molar ratio:1,15000RPM centrifuges 10 points after stirring 2 hours Clock.Latex MES（50mM, pH=6.5）Cushioning liquid cleaning is resuspended afterwards twice.Add H-FABP antibody, antibody mass and glue Newborn mass ratio is 0.04:1, stirring reaction 2 hours.Then 15000RPM is centrifuged 15 minutes, removes supernatant.Latex is in storing liquid （The trehaloses of 300+0.5% Triton X-100 of 50mM MES pH6.5+1%BSA+0.1%ProClin+2%+ 1.8% sodium chloride）Simultaneously ultrasonic disperse is resuspended, it is standby for 0.15% or so to be diluted to latex concentration.

2）The preparation of standard items

Commercially available H-FABP antigen standards are configured to series of standards product with standard dilutions, concentration is respectively 0ng/ ML, 5ng/mL, 10ng/mL, 30ng/mL, 60ng/mL, 120ng/mL.

3）The preparation of quality-control product

By the H-FABP samples of high concentration（85ng/mL）6.5ng/mL and 40ng/mL are diluted to 50mM PBS, respectively as low It is worth quality-control product and high level quality-control product.

4）Calibration curve is established in biochemical instruments

By 6 H-FABP standard items, concentration is put in biochemical instruments from 0 to 120ng/mL（Auspicious full automatic biochemical apparatus advanced in years is used in this example BS-400）In middle sample disc, the H-FABP emulsion reagents with preparation react respectively, are detected, and record degree of reaction.Detection wavelength For 700nm, using two point rate assay, reaction time 150s, 2 degree of reaction differences are calculated.Can according to concentration and degree of reaction difference To establish calibration curve.Fig. 1 is the calibration curve that emulsion reagent is established in embodiment 1.

Embodiment 2

1）The preparation of emulsion reagent

By particle diameter for 333nm surface carboxyl groups microballoon in MES（50mM, pH=6.5）1% is diluted in cushioning liquid, at room temperature Stirring.EDC solid powders are put into, it is 30 to keep EDC and carboxyl molar ratio:1,10000RPM centrifuges 10 points after stirring 2 hours Clock.Latex MES（50mM, pH=6.5）Cushioning liquid cleaning is resuspended afterwards twice.Add H-FABP antibody, antibody mass and glue Newborn mass ratio is 0.04:1, stirring reaction 2 hours.Then 10000RPM is centrifuged 15 minutes, removes supernatant.Add storing liquid （The trehaloses of 300+0.5% Triton X-100 of 50mM MES pH6.5+1%BSA+0.1%ProClin+2%+ 1.8% sodium chloride）Simultaneously ultrasonic disperse is resuspended, it is standby for 0.03% or so to be diluted to latex concentration.

2）The preparation of standard items

3）The preparation of quality-control product

4）Calibration curve is established in POCT equipment

By 6 H-FABP standard items, concentration is from 0 to 120ng/mL, and the H-FABP emulsion reagents with preparation are in reaction cup respectively Reaction, with POCT equipment（Laura electronic semiautomatic specific protein analyzer EZ-400 is used in this example）Detected, record is anti- Response.Detection wavelength is 650nm, detects scattered light intensity, using two point rate assay, reaction time 150s, calculates 2 points of reactions Spend difference.Calibration curve can be established according to concentration and degree of reaction difference.Fig. 2 is the calibration that emulsion reagent is established in embodiment 2 Curve.

Embodiment 3

The sensitivity for analysis of reagent（Blank limits）

Using the reagent in embodiment 1, tested from value standard product as dummy, in automatic clinical chemistry analyzer（Step Auspicious BS-400）On, 37 DEG C of temperature, 700nm wavelength, under the conditions of 1cm optical paths, test absorbance.Retest 20 times, according to The standard curve established in embodiment 1 calculates the average value of 20 test results（X）And standard deviation（SD）, calculate X+2SD conducts The sensitivity for analysis of reagent.Test result is as shown in table 1, and the sensitivity for analysis for showing the reagent is 0.16ng/mL.Cardioid fat Acid binding protein（H-FABP）Detection clinical reference value it is small in 5ng/mL or so, the remolding sensitivity reference value of reagent of the present invention An order of magnitude, comply fully with the needs used.

The reagent analysis sensitivity test result of the embodiment 1 of table 1.

Embodiment 1
		Test sequence number	Measurement result（ng/mL）
1	0.12
		2	0.10
3	0.06
		4	0.06
5	0.04
		6	0.02
7	0.07
		8	0.11
9	0.02
		10	0.07
11	0.11
		12	0.12
13	0.06
		14	0.05
15	0.12
		16	0.15
17	0.08
		18	0.02
19	0.15
		20	0.03
Average value（X）	0.0780
		SD	0.0423
X+2SD	0.1625

Embodiment 4

The range of linearity of reagent

With null value H-FABP standard items diluted concentrations 120ng/mL high concentration H-FABP standard items, be respectively prepared 120ng/mL, 60ng/mL, 30ng/mL, 10ng/mL, 5ng/mL, 0ng/mL totally 6 different diluted concentration samples.Using in Examples 1 and 2 The reagent of preparation is tested these samples respectively, and each diluted concentration is tested 3 times, obtains each diluted concentration testing result Average.Measurement result is shown in Table 2.Using diluted concentration as independent variable, linear regression side is obtained using testing result average as dependent variable Journey, calculate the coefficient correlation of linear regression（r）.Linear correlation curve is shown in Fig. 3 and Fig. 4.

The range of linearity measurement result of the embodiment 1 of table 2. and embodiment 2

Theoretical concentration（mg/L）	300	200	150	75	30	15
							Embodiment 1	Test value 1	303.59	209.28	152.70	78.99	29.55	15.37
Test value 2	297.96	194.95	153.92	73.97	30.64	14.51
								Test value 3	309.41	190.13	149.14	77.26	29.50	15.76
Test value average	303.65	198.12	151.92	76.74	29.90	15.21
							Embodiment 2	Test value 1	290.48	199.18	146.80	77.82	29.52	16.44
Test value 2	290.48	203.48	144.78	72.51	31.35	14.90
								Test value 3	290.48	200.74	142.99	75.06	29.10	16.40
Test value average	300.80	201.13	144.86	75.13	29.99	15.91

Embodiment 5

The repeatability of reagent

The horizontal control material of normal value for being 2.5ng/mL or so with the reagent test H-FABP concentration prepared in embodiment 2 and The horizontal control material of exceptional value that H-FABP concentration is 15ng/mL or so, each retest 10 times, calculates being averaged for measured value Value（X）And standard deviation（SD）.By formula（1）Calculate the coefficient of variation（CV）.Test result is shown in Table 3.According to measurement result, change is calculated Different coefficient CV=7.29% and 5.09%, meets reagent C V<10% technical requirements.

CV =（SD/X）×100% …………………………(1)

In formula：

CV --- the coefficient of variation；

SD --- standard deviation；

The average value of X --- measured value.

The repetition measurement result of the embodiment 2 of table 3.

Embodiment 2
			Determine sequence number	Measurement result（2.5ng/mL）	Measurement result（15ng/mL）
1	2.23	15.49
			2	2.48	14.96
3	2.45	15.23
			4	2.55	15.40
5	2.22	14.36
			6	2.53	14.95
7	2.38	15.16
			8	2.44	14.52
9	2.49	15.46
			10	2.74	15.33
Average value（X）	2.451	15.086
			SD	0.1525	0.3900
CV	6.22%	2.59%

Embodiment 6

With the correlation analysis of RANDOX reagents

In the present embodiment, we have collected 105 clinical serum samples, be examined using the reagent in embodiment 2 and equipment Survey, testing result and RANDOX reagent testing result have carried out correlation analysis.Analysis result is as shown in Figure 5.Correlation is straight The result of line fitting shows that fit equation is y=1.0022x+0.1545, coefficient R=0.9814.This result shows this hair The reagent correlation of reagent and RANDOX in bright is fine, and accuracy is very consistent.

Embodiment 7

Contrasted with the performance parameter of in the market other reagents

Performance parameter of the reagent of the present invention with the market other H-FABP reagents in POCT equipment contrasts situation as listed by table 4. As can be seen from the table, reagent of the invention mainly detection sample type, need amount of reagent and species, reagent it is linear Scope and the detection speed of reagent etc. are substantially better than the contrast agent of in the market.

The H-FABP reagent performance parameter comparisons of table 4.

Manufacturer	Health and suitable medical treatment	Import lists reagent	Domestic listing reagent
				Method of testing	Scattered light urbidmetry	Turbidimetry	Turbidimetry
Sample type	Fresh serum, blood plasma and whole blood	Fresh serum, blood plasma	Fresh serum
				Types of agents	Liquid, granules of polystyrene	Liquid, granules of polystyrene	Liquid, granules of polystyrene
Amount of reagent	1	2	2
				Reagent type	Single reagent latex	Latex and auxiliary reagent	Latex and auxiliary reagent
The range of linearity	2.5-120ng/mL	0.747-120ng/mL	2.5-160ng/mL
				Detection is time-consuming	About 2.5min	About 11min	About 10min
Minimum detectability	1ng/mL	Do not indicate	Do not indicate
				Precision	≤10%	≤10%	≤10%
The degree of accuracy	≤15%	Do not indicate	≤15%
				Difference between batch	≤10%	Do not indicate	≤10%
Whether need to be equipped with dilution	It is no	It is	It is
				The term of validity	12 months	12 months	12 months

Claims

A kind of 1. cardic fatty acid binding protein（H-FABP）Latex enhancing immune is than turbid reagent, it is characterised in that the H- FABP latex enhancing immunes are single reagent than turbid reagent, and it is molten that the reagent includes the latex particle of H-FABP antibody labelings, buffering Liquid, surfactant, salt, stabilizer, suspending agent and preservative.
2. the H-FABP latex enhancing immunes according to claim 1 are than turbid reagent, it is characterised in that described antibody is How anti-or pairing the monoclonal antibody of cardic fatty acid binding protein.
3. the H-FABP latex enhancing immunes according to claim 1 are than turbid reagent, it is characterised in that the latex particle A diameter of 150-500nm;Its surface modification group is that-COOH ,-NH2 ,-SH ,-OH, one kind in-CHO or physics are inhaled Attached type latex particle；Its surface is by H-FABP antibody labelings；The concentration of latex particle is 0.01%-1%.
4. the H-FABP latex enhancing immunes according to claim 1 are than turbid detection reagent, it is characterised in that the stabilization Agent is one kind in bovine serum albumin(BSA), casein, gelatin；The salt is in sodium chloride, potassium chloride, sodium sulphate, potassium sulfate A kind of, two or more combination；Described surfactant is NP40, triton x-100, polysorbas20, polysorbate40, is told Temperature 80, lauryl sodium sulfate（SDS）, span 40, one kind in sorbester p18, two or more combination；The suspending Agent be sucrose, glucose, maltose, trehalose, mannitol, ethylene glycol, glycerine, one kind in lactose, two kinds or two kinds with On combination；Described preservative is one kind in sodium azide, thimerosal, Proclin-300;The cushioning liquid is sweet Propylhomoserin buffer solution, 4- hydroxyethyl piperazineethanesulfonic acids（HEPES）Buffer solution, 3- (N- morpholines) -2- hydroxy-propanesulfonic acids（MOPSO）Buffering Liquid, trihydroxy aminomethane hydrochloric acid（Tris-HCl）Buffer solution, phosphoric acid physiological saline（PBS）Buffer solution, MES （MES）One kind in buffer solution.
5. the H-FABP latex enhancing immunes according to claim 5 are than turbid detection reagent, it is characterised in that the stabilization Agent is bovine serum albumin(BSA), one kind in casein, gelatin, concentration 0.2%-2%.
6. H-FABP latex enhancing immunes according to claim 5 are than turbid detection reagent, it is characterised in that the suspending agent For one kind in sucrose, glucose, maltose, trehalose, mannitol, ethylene glycol, glycerine, lactose, two or more Combination, concentration 0.2%-5%.
7. the H-FABP latex enhancing immunes according to claim 5 are than turbid detection reagent, it is characterised in that the buffering Solution is glycine buffer, 4- hydroxyethyl piperazineethanesulfonic acids（HEPES）Buffer solution, 3- (N- morpholines) -2- hydroxy-propanesulfonic acids （MOPSO）Buffer solution, trihydroxy aminomethane hydrochloric acid（Tris-HCl）Buffer solution, phosphoric acid physiological saline（PBS）Buffer solution, 2- Quinoline ethyl sulfonic acid（MES）One kind in buffer solution, concentration range 10-200mmol/L, pH scope is between 5.5-9.0.
8. H-FABP latex enhancing immunes according to claim 5 are than turbid detection reagent, it is characterised in that the salt is One kind, two or more combination in sodium chloride, potassium chloride, sodium sulphate, potassium sulfate, concentration 0.5%-3.5%.
9. H-FABP latex enhancing immunes according to claim 5 are than turbid detection reagent, it is characterised in that the anti-corrosion Agent is sodium azide, one kind in thimerosal, Proclin-300, concentration 0.05%-2%.
10. H-FABP latex enhancing immunes according to claim 5 are than turbid detection reagent, it is characterised in that the surface Activating agent is NP40, triton x-100, polysorbas20, polysorbate40, Tween 80, lauryl sodium sulfate（SDS）, span 40, sapn One kind, two or more combination in 60, concentration 0.1%-2%.
11. a kind of latex enhancing immune detects cardic fatty acid binding protein than turbid single reagent（H-FABP）Method, its feature It is, using any H-FABP latex enhancing immunes described in claim 1-10 than turbid reagent, in Biochemical Analyzer Either test sample is treated in the equipment such as specific protein analyzer by detecting the method for transmitted light or scattered light intensity change to determine H-FABP contents in product.
12. detection method according to claim 12, it is characterised in that used detection sample can be whole blood, serum Or blood plasma.