| There are more than 100.000 kinds of proteins in human body.which assume a different task.In order to reveal the laws of life activities.it is an inevitable direction of development which identify and detect those proteins and their physiological functions.They come from rhagiocrine cell,and play an important role in sustaining the metergasis of human bodies.Those proteins which were called Specific protein play an important pathological significance in a state of disease.And the body's immune system also exist in a particular type of protein,they aer Closely related with the occurrence of disease of immune system,development.turnover and prognosis.including immunoglobulin A,immunoglobulin G and immunoglobulin M. as well as complement 3,complement 4.The body's immune system is a Self-defense defense system.which can maintain human health,and prevent the occurrence of various kinds of diseases,it is gradual establishment and perfection in the human evolutionary process,and in fighting against the natural environment,is a well-organized,the sophisticated of division of labour,When the disease,resistant to ensure that the unity of the human body environment,stability and balance.The human body immune dysfunction will cause cancer,chronic kidney disease,prostate. chronic hepatitis,chronic gastropathy,diabetes,lupus erythematosus,rheumatic heart disease,coronary heart disease,hypertension,sclerosis,myasthenia gravis. rheumatoid,senile diseases such as autoimmune diseases,which seriously threat to human health.With the development of society and the continuous advancement of medical technology,more and more people immune system disease provoke people's attention.At present,the specific protein commonly used detection methods, including Single Radial Immunodiffusion(SRID),Immunoelectrophoresis, Enzyme-linked Immunosorbent Assay(ELISA).Chemiluminescence Immunoassay (CLIA),Time-resolved Immunoassay(TRFIA),Transmission Turbidity te al.but these methods which are some drawbacks,such as operation cumbersome, time-consuming and difficult to automate,so when testing which will not only increase the financial burden,but also spend more of time,energy and reagents.Along with the development of experimental technique Special protein analysis technology which from the early test-tube precipitation reactions and agar gel diffusion test.develope to today's immunoassay technology,the special protein immunological analysis technique gradual perfect,sensitivity gradually increased. detection levels by micrograms(μg) to the development of Tenneco(ng),even Pique (pg) level.Special protein analyzer is often purchased in the domestic,mainly for the clinical detection of special test equipment.Specific protein analysis useing principle which is that antigen - antibodies response under certain conditions to form immune complexes.Immunoturbidimetry with the rapid detection of biochemical and immune analysis of the specificity,is the modern optical measuring instruments and automation inspection system of combining the methods used in reaction sediments may be all kinds of liquids in micro - Antigen,antibody and small molecule drugs and semi-quantitative determination of antigen material.And other immune analysis method,in addition to immune turbidity technology is simple,flexible,and testing a wide range of automation and so easy striking feature of its most prominent advantages to overcome the antigen is also caused by excessive high-dose HOOK effect.The earliest dating back to 1967,Ritchie and other scattered reports of the turbidity(nephelometry),its principle is the detection of light through the solution, which was contained in the antigen-antibody complex refraction and some deflection, a scattering light.According to Raleigh scattering formula,under certain conditions,transmission and inversely proportional to the concentration of particulate,the scattered lightisproportional to the concentration of particles.Scattering turbidity can be divided into terminal and the rate of scattering of Nephelometry timing and dynamic scattering turbidity.Immune scattering turbidimetry method,with a wide range of linear,the determination of speed,precision,specificity,repeatable,stable and reliable results,and the advantages of bulk,is a trace,rapid,automated testing of body fluids Specific protein components of the immune chemical analysis technique.In recent years,the domestic clinical testing with the increased demand. immunization turbidity determination of the application of more and more common. corresponding to the use of reagents were also broader trends,but because the domestic use of the basic equipment dependent on imports,which led to the A serious problem,most of these machines can only use their own original reagents and buffer, they are most expensive,unsuitable for long-term use of large quantities to the clinical use of limitations.If the equipment can be developed with the support of reagents and buffer,clinical testing will solve a major problem.At present the domestic companies have this kind of research,but the results are not ideal and remain in the initial stage.Therefore supporting the development of China-made reagents and related buffer has an enormous market prospects.China will not only fill the vacancy reagents,and can promote domestic clinical testing technology development and progress.Given the clinical detection of specific protein and the need for the limitations of current detection methods,our purpose is to develop immunity to be applied scattering turbidity technology related to the domestic detection reagents and buffer to meet domestic market demand and Majority of patients with clinical bring the gospel.The laboratory is used regularly dynamic scattering turbid metric method,the principle of this method:As immunoprecipitation in the antigen-antibody reaction is to begin immediately after the encounter,in a very short time in the reaction medium scattering signal changing a lot at this time of peak signal will produce a certain error. Therefore.in response to antigen-antibody reaction time left PWC,scattering light signal at the beginning of the first reading of 7.5 s～2min reaction,in most cases 2 min after the second reading test.and the second reading from the first reading deducted and finally converted to analyte concentration of antigen.Heidel Berger found 1 antigen antibody dose - response curve.At Antibody excessive stage,the content of sediments with the antigen concentration ratio increased,during the increase in concentration when antigen is no longer to enhance the signal,they reached a point of balance.Antigen concentration will lead to further increase of precipitation reduced the total.When the antigen concentration far exceeds all antibodies binding capacity,it will produce soluble material,and then each other cross linking formation of sediments can not be detected.Excessive antigen in the region,all have greater concentration of pathology in human proteins,are subject to the impact of this phenomenon.According to the shape of the response curve,Green and the others proposed a "hook effect(hook effect)" name.At present,the hook effect mainly described antigen surplus,we use the special protein-BN Prospec has advantages as follow:First.the timing dynamic scattering turbidity,the former scattering of light to the whole collection and latex reagents to increase ten-fold increase in sensitivity.Second,the surplus equipment also has automatic dilution antigen detection,indirect excessive antigen detection,to avoid the effect of high-dose Hook false negative results of the detection of a broader scope.In the experiment we antigen-antibody reaction principle of dynamic timing analysis technology scattering turbidity monitoring serum immunoglobulin in the system of the associated protein content,in order to evaluate the occurrence of clinical disease, development,as well as the vesting of the prognosis.Evaluation of the methods of precision,sensitivity,stability and normal reference values,and other indicators,and with companies with similar Dabe Behring reagents for the detection of serum samples related to the outcome of the comparison.Measurement results of the relevant comparison.The results showed:(1) the reaction of imports and dilution buffer and the reaction solution developed by the laboratory and dilution under the same conditions were normal in 30 specimens were compared results of a good correlation,r = 0.991,P＜0.01.line of the equation Y(self-made reagent) =- 0.11 +1.012 X(Germany Ling reagent),as a result,significant statistical significance, repeatability,stability,and 37℃for next year are kept stable effective.(2)IgA,IgG, IgM reagents:the minimum detection limit for 0.028 g·L-1,0.72 g·L-1,0.022 g·L-1;approved the precision of 2.1%- 4.9%,1.6%to 3.5%,3.5%to 5.0%,the precision of 2.0%to 3.1%.3.0%to 4.5%,2.6%to 4.1%;IgA samples of free hemoglobin,bilirubin,triglyceride concentration of interference 10 g·L-1,600mg·L-1,9.9 g·L-1,the test results had no significant effects of IgG in the sample of free hemoglobin,bilirubin,triglyceride concentration interference,respectively 10 g·L-1, 600mg·L-1,19 g·L-1 test results when there was no significant impact,IgM samples of free hemoglobin,bilirubin,triglyceride concentration interference, respectively 10 g·L-1,600mg·L-1,4.6 g·L-1,the test results had no significant impact;Detection of 54 serum samples,and imports of similar reagents,the results show a good correlation between the two reagents,as a result,significant statistical significance:IgA linear equation Y(self-made reagent) = 1.101 +0.881 X(Germany Ling reagent),a correlation coefficient of 0.980;IgG linear equation Y(self-made reagent) = 0.148 +0.901 X(Germany Ling reagent),the correlation coefficient was 0.979:IgM linear equation Y(self-made reagent) =1.045 -0.075X(Germany Ling reagent ),the correlation coefficient was 0.898.Normal reference values of IgA,IgG, IgM were 0.59 to 4.31 g·L-1,6.9 to 17.92 g·L-1.0.35 to 2.23 g·L-1.(3) C3 and C4:the minimum detection limit for 0.022 g·L-1,0.0014 g·L-1:approved the precision of 2.0%- 4.9%,4.5%to 5.0%.the precision of 5.4%- 6.8%.3.8%to 5.1% C3 samples of free hemoglobin,bilirubin,triglyceride concentration interference, respectively 10 g·L-1,600mg·L-1.5.7 g·L-1 on the test results had no significant impact,C4 samples of free hemoglobin,bilirubin,interference triglyceride concentrations were 10 g·L-1.600mg·L-1.2.4 g·L-1 when test results without significant impact on the detection of 54 serum samples,and imports of similar reagents,the results show a good correlation between the two reagents,as a result, significant statistical significance:C3 linear equation Y(self-made reagent) =- 5.71 -5E +1.006 X(Germany Ling reagent),the correlation coefficient was 0.974;C4 linear equation Y(self-made reagent) = 1.101 +0.993 X(Germany Ling reagent) with a correlation coefficient of 0.961.Normal reference range C3,C4 were 0.90 - 1.68 g·L-1,0.110 - 0.408g·L- 1.(4)Stability:immunoglobulins immune antibody reagents and turbidity standard at 37℃for seven days,scattering light Value standard value has dropped but a good standard curve,no change detection range,three positive Detection of serum and placed before the close,and within the approved grant of precision CV remains below 10%.Our results showed that the timing scattering turbidity analysis of immunoglobulin system IgA,IgG.IgM,as well as C3,C4 good repeatability,high accuracy,high sensitivity,and the Dade Behringd compared with the results of similar reagents,a good correlation,and simple,short reaction time,as well as a large quantity of samples can achieve the advantages of automation,as a supplementary means of diagnosis has great potential for clinical application.The clinical application for the further development of special protein detection reagents laid a solid foundation. |
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