技术领域technical field
本发明涉及体外诊断试剂技术领域，特别涉及一种脂蛋白(a)的检测试剂及方法。The invention relates to the technical field of in vitro diagnostic reagents, in particular to a detection reagent and method for lipoprotein (a).
背景技术Background technique
脂蛋白(a)是一种特殊的血浆脂蛋白。在1975年Dalhen等研究认为Lpa是动脉粥样硬化的危险因子。1978年，Mclean等研究脂蛋白(a)中的Apo(a)与纤溶酶原(PLG)具有高度同源性，从而认为脂蛋白(a)不仅是动脉粥样硬化的危险因素，而且可能与纤溶系统有关。脂蛋白(a)在预测动脉粥样硬化性心血管疾病的发生、冠状动脉粥样硬化性心脏病(冠心病)的再发、静脉血栓事件的发生以及肾脏移植预后等方面均有重要的临床价值。Lipoprotein(a) is a special type of plasma lipoprotein. In 1975, Dalhen et al. considered that Lpa was a risk factor for atherosclerosis. In 1978, Mclean et al. studied that Apo(a) in lipoprotein(a) has a high degree of homology with plasminogen (PLG), so that lipoprotein(a) is not only a risk factor for atherosclerosis, but may also be a risk factor for atherosclerosis. related to the fibrinolytic system. Lipoprotein (a) has important clinical significance in predicting the occurrence of atherosclerotic cardiovascular disease, the recurrence of coronary atherosclerotic heart disease (CHD), the occurrence of venous thrombotic events, and the prognosis of kidney transplantation. value.
早期检测脂蛋白(a)所用的电泳法灵敏度低，多用于定性检测。随后研发出了多种能直接测定脂蛋白(a)的免疫化学检测法，如辐射状免疫扩散法(RID)、电免疫扩散法(EID)、放射免疫测定法(RIA)、酶联免疫吸附试验(ELISA)、免疫浊度法[包括免疫散射比浊法(INA)和免疫透射比浊法(ITA)]、解离增强配体荧光免疫测定法(DELFIA)等。RID法与EID法因操纵简便，不需特殊设备，仍有一些基层单位实验室采用，但缺点是灵敏度低。RIA法的缺点是操纵复杂，有放射性核素污染。DELFIA法因需特殊仪器，国内也较少应用。其中免疫比浊法，快速简便、精密度高、易于自动化、适于大批量标本的同时检测，便于推广，适合临床检测。但需要的抗体用量大，对抗体要求高(应具有高特异性、高滴度和高亲和力)，颗粒大小不同的Lp(a)会产生不一致的光散射与光吸收，而且受标本中的基质的影响较明显，容易受到类风湿因子的干扰，所以研发一种成本低、特异性好、偏差小的检测方法是目前的市场需求。The electrophoresis method used for early detection of lipoprotein (a) has low sensitivity and is mostly used for qualitative detection. Subsequently, a variety of immunochemical assays that can directly measure lipoprotein(a) have been developed, such as radial immunodiffusion (RID), electric immunodiffusion (EID), radioimmunoassay (RIA), enzyme-linked immunosorbent assay Assay (ELISA), immunoturbidimetric assay [including immune nephelometry (INA) and immunotransmission turbidimetry (ITA)], dissociation-enhancing ligand fluorescence immunoassay (DELFIA), etc. RID method and EID method are still used in some basic unit laboratories because of their simple operation and no need for special equipment, but the disadvantage is that the sensitivity is low. The disadvantage of the RIA method is that it is complicated to operate and contaminated with radionuclides. Because DELFIA method requires special equipment, it is rarely used in China. Among them, the immunoturbidimetric method is quick and simple, has high precision, is easy to automate, is suitable for simultaneous detection of a large number of specimens, is easy to popularize, and is suitable for clinical detection. However, the amount of antibody required is large, and the antibody requirements are high (should have high specificity, high titer and high affinity). Lp(a) with different particle sizes will produce inconsistent light scattering and light absorption, and it is affected by the matrix in the specimen. The influence of rheumatoid factor is obvious, and it is easy to be interfered by rheumatoid factors. Therefore, it is the current market demand to develop a detection method with low cost, good specificity and small deviation.
公开号为CN103604930A中公开了一种脂蛋白(a)的检测试剂盒，组成包括：Tris-HCl缓冲液，脂蛋白(a)100mg/dL，氯化钠150mM，蔗糖5.0％，BSA1.0％，EDTA10mM，叠氮化钠0.1％。但该试剂盒在抗干扰能力和精密度方面还存在一定的欠缺。Publication No. CN103604930A discloses a detection kit for lipoprotein (a), the composition includes: Tris-HCl buffer, lipoprotein (a) 100mg/dL, sodium chloride 150mM, sucrose 5.0%, BSA 1.0% , EDTA 10mM, sodium azide 0.1%. However, the kit still has some deficiencies in anti-interference ability and precision.
发明内容SUMMARY OF THE INVENTION
有鉴于此，本发明提供了一种脂蛋白(a)的检测试剂及方法。该检测试剂及方法抗干扰能力强、特异性高、准确度高、稳定性好。In view of this, the present invention provides a detection reagent and method for lipoprotein (a). The detection reagent and method have strong anti-interference ability, high specificity, high accuracy and good stability.
为了实现上述发明目的，本发明提供以下技术方案：In order to achieve the above-mentioned purpose of the invention, the present invention provides the following technical solutions:
本发明提供了一种脂蛋白(a)的检测试剂，该检测试剂由试剂1和试剂2组成；The present invention provides a detection reagent for lipoprotein (a), the detection reagent is composed of reagent 1 and reagent 2;
其中，试剂1包括MOPS缓冲液、Tween80、防腐剂和水；Wherein, reagent 1 includes MOPS buffer, Tween80, preservative and water;
试剂2包括包被有脂蛋白(a)单克隆抗体的胶乳微球、甘氨酸缓冲液、Tween80、防腐剂、甘油和水。Reagent 2 includes latex microspheres coated with lipoprotein(a) monoclonal antibody, glycine buffer, Tween 80, preservative, glycerol and water.
作为优选，试剂1中各组分的用量为：Preferably, the consumption of each component in Reagent 1 is:
MOPS缓冲液：50～200mmol/L；MOPS buffer: 50～200mmol/L;
Tween80：0.5～5vt％；Tween80: 0.5～5vt%;
防腐剂：0.05～0.2wt％；Preservative: 0.05～0.2wt%;
优选地，试剂1中各组分的用量为：Preferably, the dosage of each component in Reagent 1 is:
MOPS缓冲液：50～100mmol/L；MOPS buffer: 50～100mmol/L;
Tween80：1～3vt％；Tween80: 1～3vt%;
防腐剂：0.05wt％。Preservative: 0.05 wt%.
优选地，试剂1中Tween80的浓度为1～2vt％。Preferably, the concentration of Tween80 in Reagent 1 is 1-2 vt%.
在本发明提供的实施例中，试剂1中各组分的用量为：In the embodiment provided by the present invention, the consumption of each component in Reagent 1 is:
MOPS缓冲液：100mmol/L；MOPS buffer: 100mmol/L;
Tween80：1.5vt％；Tween80: 1.5vt%;
防腐剂：0.05wt％。Preservative: 0.05 wt%.
作为优选，试剂1的pH值为6～8。Preferably, the pH value of reagent 1 is 6-8.
优选地，试剂1的pH值为6.5～7.5。Preferably, the pH value of reagent 1 is 6.5-7.5.
在本发明提供的实施例中，试剂1的pH值为7.0。In the examples provided by the present invention, the pH value of reagent 1 is 7.0.
作为优选，试剂2中各组分的用量为：Preferably, the consumption of each component in Reagent 2 is:
包被有脂蛋白(a)单克隆抗体的胶乳微球：0.001～0.02g/L；Latex microspheres coated with lipoprotein (a) monoclonal antibody: 0.001～0.02g/L;
甘氨酸缓冲液：10～100mmol/L；Glycine buffer: 10～100mmol/L;
Tween80：0.5～5vt％；Tween80: 0.5～5vt%;
防腐剂：0.05～0.2wt％；Preservative: 0.05～0.2wt%;
甘油：2～10vt％；Glycerol: 2～10vt%;
优选地，试剂2中各组分的用量为：Preferably, the consumption of each component in Reagent 2 is:
包被有脂蛋白(a)单克隆抗体的胶乳微球：0.001～0.01g/L；Latex microspheres coated with lipoprotein (a) monoclonal antibody: 0.001～0.01g/L;
甘氨酸缓冲液：10～50mmol/L；Glycine buffer: 10～50mmol/L;
Tween80：1～5vt％；Tween80: 1～5vt%;
防腐剂：0.1wt％；Preservative: 0.1wt%;
甘油：2～10vt％。Glycerol: 2～10vt%.
优选地，试剂2中Tween80的浓度为2～5vt％。Preferably, the concentration of Tween80 in Reagent 2 is 2-5 vt%.
在本发明提供的实施例中，试剂2中各组分的用量为：In the embodiment provided by the invention, the consumption of each component in reagent 2 is:
包被有脂蛋白(a)单克隆抗体的胶乳微球：0.001g/L；Latex microspheres coated with lipoprotein (a) monoclonal antibody: 0.001g/L;
甘氨酸缓冲液：50mmol/L；Glycine buffer: 50mmol/L;
Tween80：2vt％；Tween80: 2vt%;
防腐剂：0.1wt％；Preservative: 0.1wt%;
甘油：3vt％。Glycerol: 3vt%.
作为优选，试剂2的pH值为7～9。Preferably, the pH value of the reagent 2 is 7-9.
优选地，试剂2的pH值为7～8。Preferably, the pH value of reagent 2 is 7-8.
在本发明提供的实施例中，试剂2的pH值为7.5。In the examples provided by the present invention, the pH value of reagent 2 is 7.5.
在本发明中，适宜pH的选择提高了试剂的贮存稳定性。In the present invention, the selection of a suitable pH improves the storage stability of the reagent.
作为优选，防腐剂为叠氮钠。Preferably, the preservative is sodium azide.
在本发明提供的实施例中，包被有脂蛋白(a)单克隆抗体的胶乳微球是由脂蛋白(a)单克隆抗体和胶乳微球采用化学交联法得到。In the examples provided by the present invention, the latex microspheres coated with the lipoprotein (a) monoclonal antibody are obtained by chemical cross-linking between the lipoprotein (a) monoclonal antibody and the latex microspheres.
在本发明提供的实施例中，试剂2中包被有脂蛋白(a)单克隆抗体的胶乳微球的制备方法为：将脂蛋白(a)单克隆抗体与经过MES活化的胶乳微球混合，30℃交联得到。In the examples provided by the present invention, the preparation method of the latex microspheres coated with the lipoprotein (a) monoclonal antibody in the reagent 2 is: mixing the lipoprotein (a) monoclonal antibody and the latex microspheres activated by MES , obtained by crosslinking at 30°C.
作为优选，脂蛋白(a)单克隆抗体为鼠抗人载脂蛋白(a)单克隆抗体。Preferably, the lipoprotein (a) monoclonal antibody is a mouse anti-human apolipoprotein (a) monoclonal antibody.
作为优选，胶乳微球为疏水微球。胶乳微球的选择提高了化学交联的效率，减少了抗体的损失，降低了成本，提高了灵敏度和精密度。Preferably, the latex microspheres are hydrophobic microspheres. The choice of latex microspheres increases the efficiency of chemical cross-linking, reduces antibody loss, reduces cost, and improves sensitivity and precision.
作为优选，疏水微球为带磺酸基基团、羧基基团或醛基基团的一种或两种以上的胶乳微球。Preferably, the hydrophobic microspheres are one or more latex microspheres with sulfonic acid group, carboxyl group or aldehyde group.
优选地，疏水微球为带磺酸基基团和/或羧基基团的胶乳微球。Preferably, the hydrophobic microspheres are latex microspheres with sulfonic acid groups and/or carboxyl groups.
作为优选，胶乳微球的平均粒径为100～300nm。Preferably, the average particle size of the latex microspheres is 100-300 nm.
优选地，胶乳微球的平均粒径为200～300nm。Preferably, the average particle size of the latex microspheres is 200-300 nm.
作为优选，脂蛋白(a)单克隆抗体与胶乳微球的质量比为(0.5～1)：100。Preferably, the mass ratio of the lipoprotein (a) monoclonal antibody to the latex microspheres is (0.5-1):100.
优选地，脂蛋白(a)单克隆抗体与胶乳微球的质量比为0.8：100。Preferably, the mass ratio of lipoprotein (a) monoclonal antibody to latex microspheres is 0.8:100.
本发明还提供了一种脂蛋白(a)含量的检测方法，包括：待测样本加入试剂1中，孵育5～10min后再加入试剂2形成凝集，在660nm波长下测定吸光度，通过脂蛋白(a)的标准曲线得到待测样本中脂蛋白(a)的含量；The present invention also provides a method for detecting the content of lipoprotein (a), comprising: adding a sample to be tested into reagent 1, incubating for 5-10 minutes and then adding reagent 2 to form agglutination, measuring absorbance at a wavelength of 660 nm, and using lipoprotein ( The standard curve of a) obtains the content of lipoprotein (a) in the sample to be tested;
其中，试剂1包括MOPS缓冲液、Tween80、防腐剂和水；Wherein, reagent 1 includes MOPS buffer, Tween80, preservative and water;
试剂2包括包被有脂蛋白(a)单克隆抗体的胶乳微球、甘氨酸缓冲液、Tween80、防腐剂、甘油和水。Reagent 2 includes latex microspheres coated with lipoprotein(a) monoclonal antibody, glycine buffer, Tween 80, preservative, glycerol and water.
作为优选，孵育的温度为37℃。Preferably, the incubation temperature is 37°C.
作为优选，在检测过程中试剂1与试剂2的体积比为3:1。Preferably, in the detection process, the volume ratio of reagent 1 to reagent 2 is 3:1.
本发明提供了一种脂蛋白(a)的检测试剂及方法。该检测试剂由试剂1和试剂2组成；其中，试剂1包括MOPS缓冲液、Tween80、防腐剂和水；试剂2包括包被有脂蛋白(a)单克隆抗体的胶乳微球、甘氨酸缓冲液、Tween80、防腐剂、甘油和水。本发明至少具有如下优势之一：The present invention provides a detection reagent and method for lipoprotein (a). The detection reagent is composed of reagent 1 and reagent 2; wherein, reagent 1 includes MOPS buffer, Tween80, preservative and water; reagent 2 includes latex microspheres coated with lipoprotein (a) monoclonal antibody, glycine buffer, Tween80, preservatives, glycerin and water. The present invention has at least one of the following advantages:
(1)本发明检测脂蛋白(a)的试剂及方法检测结果准确性高、灵敏度好、精密度好、线性范围宽，且稳定性好，保存期长，便于推广使用，成本低，可广泛应用于各级医院、卫生预防部门和医学生物科研单位测定人血清中脂蛋白(a)的含量。(1) The reagent and method for detecting lipoprotein (a) of the present invention have high detection results, high accuracy, good sensitivity, good precision, wide linear range, good stability, long shelf life, easy popularization and use, low cost, and can be widely used. It is used in the determination of lipoprotein (a) content in human serum in hospitals at all levels, health prevention departments and medical and biological scientific research units.
(2)本发明检测试剂抗干扰能力强、特异性高。表面活性剂的选择，可以减少样本的浊度对实验结果的影响；抗体的选择特异性更好，可以减少非特异性干扰和类风湿因子的干扰。(2) The detection reagent of the present invention has strong anti-interference ability and high specificity. The choice of surfactant can reduce the influence of turbidity of the sample on the experimental results; the choice of antibody is more specific, which can reduce the interference of non-specific interference and rheumatoid factor.
(3)试剂制备过程中简化了处理胶乳微球和标记抗体的时间，简化了工艺流程，使操作更简便。(3) In the process of reagent preparation, the time for processing latex microspheres and labeled antibodies is simplified, the process flow is simplified, and the operation is easier.
具体实施方式Detailed ways
本发明公开了一种脂蛋白(a)的检测试剂及方法，本领域技术人员可以借鉴本文内容，适当改进工艺参数实现。特别需要指出的是，所有类似的替换和改动对本领域技术人员来说是显而易见的，它们都被视为包括在本发明。本发明的方法及应用已经通过较佳实施例进行了描述，相关人员明显能在不脱离本发明内容、精神和范围内对本文的方法和应用进行改动或适当变更与组合，来实现和应用本发明技术。The invention discloses a detection reagent and method for lipoprotein (a), and those skilled in the art can learn from the content of this article and appropriately improve the process parameters to achieve. It should be particularly pointed out that all similar substitutions and modifications are obvious to those skilled in the art, and they are deemed to be included in the present invention. The method and application of the present invention have been described through the preferred embodiments, and it is obvious that relevant persons can make changes or appropriate changes and combinations of the methods and applications herein without departing from the content, spirit and scope of the present invention, so as to realize and apply the present invention. Invention technology.
本发明提供的脂蛋白(a)的检测试剂及方法中所用试剂或仪器均可由市场购得。The detection reagents for lipoprotein (a) provided by the present invention and the reagents or instruments used in the method can be purchased from the market.
以下实施例中市售国产脂蛋白(a)试剂盒(胶乳增强免疫比浊法)的配方如下：The formula of commercially available domestic lipoprotein (a) kit (latex-enhanced immune turbidimetry) in the following examples is as follows:
试剂1：40mmol/L含1.0％NaN3的甘氨酸缓冲液；Reagent 1: 40mmol/L glycine buffer containing 1.0% NaN3;
试剂2：抗人脂蛋白(a)-IgG的致敏乳胶颗粒悬液。Reagent 2: Sensitized latex particle suspension of anti-human lipoprotein(a)-IgG.
下面结合实施例，进一步阐述本发明：Below in conjunction with embodiment, the present invention is further elaborated:
实施例1：本发明检测脂蛋白(a)的试剂及方法Example 1: Reagent and method for detecting lipoprotein (a) of the present invention
包被有鼠抗人脂蛋白(a)单克隆抗体的胶乳颗粒的制备：Preparation of latex particles coated with mouse anti-human lipoprotein(a) monoclonal antibody:
用等量的MES溶液活化0.01％羧基基团胶乳微球，然后把5mg/mL鼠抗人脂蛋白(a)单克隆抗体，与经过活化的胶乳微球混合，30℃交联得到。0.01% carboxyl group latex microspheres were activated with an equal amount of MES solution, and then 5 mg/mL mouse anti-human lipoprotein (a) monoclonal antibody was mixed with the activated latex microspheres and cross-linked at 30°C.
本实施例试剂包括：The reagents in this example include:
试剂1：100mmol/L MOPS缓冲液，Tween80 1.2％，防腐剂叠氮钠0.05％，pH 7.0；Reagent 1: 100mmol/L MOPS buffer, Tween80 1.2%, preservative sodium azide 0.05%, pH 7.0;
试剂2：包被有鼠抗人脂蛋白(a)单克隆抗体的胶乳颗粒0.001g/L，粒径为200～300nm，50mol/L甘氨酸缓冲液，0.1％叠氮钠，2％Tween80，3％甘油，pH值7.5。Reagent 2: latex particles coated with mouse anti-human lipoprotein (a) monoclonal antibody 0.001g/L, particle size 200-300nm, 50mol/L glycine buffer, 0.1% sodium azide, 2% Tween80, 3 % Glycerol, pH 7.5.
检测方法：待测样本血清加入试剂1中，孵育5min中再加入试剂2形成凝集，试剂1与试剂2的体积比为3:1，在660nm波长下测定吸光度，通过脂蛋白(a)的标准曲线计算样品中脂蛋白(a)的含量。Detection method: The serum of the sample to be tested is added to Reagent 1, and after incubation for 5 minutes, Reagent 2 is added to form agglutination. The volume ratio of Reagent 1 and Reagent 2 is 3:1, and the absorbance is measured at a wavelength of 660 nm, through the standard of lipoprotein (a). The curve calculates the content of lipoprotein (a) in the sample.
实施例2：本发明检测脂蛋白(a)的试剂及方法Example 2: Reagent and method for detecting lipoprotein (a) of the present invention
包被有鼠抗人脂蛋白(a)单克隆抗体的胶乳颗粒的制备：Preparation of latex particles coated with mouse anti-human lipoprotein(a) monoclonal antibody:
用等量的MES溶液活化0.01％磺酸基基团胶乳微球，然后把5mg/mL鼠抗人脂蛋白(a)单克隆抗体，与经过活化的胶乳微球混合，30℃交联得到。0.01% sulfonic acid group latex microspheres were activated with an equal amount of MES solution, and then 5 mg/mL mouse anti-human lipoprotein (a) monoclonal antibody was mixed with the activated latex microspheres and cross-linked at 30°C.
本实施例试剂包括：The reagents in this example include:
试剂1：100mmol/L MOPS缓冲液，Tween80 1.2％，防腐剂叠氮钠0.05％，参考pH7.0；Reagent 1: 100mmol/L MOPS buffer, Tween80 1.2%, preservative sodium azide 0.05%, reference pH 7.0;
试剂2：包被有鼠抗人脂蛋白(a)单克隆抗体的胶乳颗粒0.001g/L，粒径为200～300nm，50mol/L甘氨酸缓冲液，0.1％叠氮钠，2％Tween80，3％甘油，pH值7.5。Reagent 2: latex particles coated with mouse anti-human lipoprotein (a) monoclonal antibody 0.001g/L, particle size 200-300nm, 50mol/L glycine buffer, 0.1% sodium azide, 2% Tween80, 3 % Glycerol, pH 7.5.
检测方法：待测样本血清加入试剂1中，孵育5min中再加入试剂2形成凝集，试剂1与试剂2的体积比为3:1，在660nm波长下测定吸光度，通过脂蛋白(a)的标准曲线计算样品中脂蛋白(a)的含量。Detection method: The serum of the sample to be tested is added to Reagent 1, and after incubation for 5 minutes, Reagent 2 is added to form agglutination. The volume ratio of Reagent 1 and Reagent 2 is 3:1. The absorbance is measured at a wavelength of 660 nm, and the standard of lipoprotein (a) is passed. The curve calculates the content of lipoprotein (a) in the sample.
实施例3：本发明检测脂蛋白(a)的试剂及方法Embodiment 3: The reagent and method of the present invention to detect lipoprotein (a)
包被有鼠抗人脂蛋白(a)单克隆抗体的胶乳颗粒的制备：Preparation of latex particles coated with mouse anti-human lipoprotein(a) monoclonal antibody:
用等量的MES溶液活化0.01％羧基和磺酸基基团胶乳微球，，然后把5mg/mL鼠抗人脂蛋白(a)单克隆抗体，与经过活化的胶乳微球混合，30℃交联得到。Activated 0.01% carboxyl and sulfonic acid group latex microspheres with an equal amount of MES solution, then mixed 5 mg/mL mouse anti-human lipoprotein (a) monoclonal antibody with the activated latex microspheres, and exchanged them at 30°C. Link to get.
本实施例试剂包括：The reagents in this example include:
试剂1：100mmol/L MOPS缓冲液，Tween80 1.2％，防腐剂叠氮钠0.05％，参考pH7.0；Reagent 1: 100mmol/L MOPS buffer, Tween80 1.2%, preservative sodium azide 0.05%, reference pH 7.0;
试剂2：包被有鼠抗人脂蛋白(a)单克隆抗体的胶乳颗粒0.001g/L，粒径为200～300nm，50mol/L甘氨酸缓冲液，0.1％叠氮钠，2％Tween80，3％甘油，pH值7.5。Reagent 2: latex particles coated with mouse anti-human lipoprotein (a) monoclonal antibody 0.001g/L, particle size 200-300nm, 50mol/L glycine buffer, 0.1% sodium azide, 2% Tween80, 3 % Glycerol, pH 7.5.
检测方法：待测样本血清加入试剂1中，孵育5min中再加入试剂2形成凝集，试剂1与试剂2的体积比为3:1，在660nm波长下测定吸光度，通过脂蛋白(a)的标准曲线计算样品中脂蛋白(a)的含量。Detection method: The serum of the sample to be tested is added to Reagent 1, and after incubation for 5 minutes, Reagent 2 is added to form agglutination. The volume ratio of Reagent 1 and Reagent 2 is 3:1. The absorbance is measured at a wavelength of 660 nm, and the standard of lipoprotein (a) is passed. The curve calculates the amount of lipoprotein (a) in the sample.
实施例4：本发明检测方法的准确度分析Embodiment 4: the accuracy analysis of the detection method of the present invention
试验仪器：日立7080全自动生化分析仪；Test instrument: Hitachi 7080 automatic biochemical analyzer;
检测样品：20名体检者血清样本；Test samples: Serum samples from 20 medical examiners;
对比试剂1：市售国产脂蛋白(a)试剂盒(胶乳增强免疫比浊法)。Comparative reagent 1: commercially available domestic lipoprotein (a) kit (latex-enhanced immune turbidimetry).
用实施例1至实施例3的检测试剂和对比试剂分别测定20例样品，检测结果见表1-1～1-3。Twenty samples were measured with the detection reagents and comparative reagents of Examples 1 to 3, respectively, and the detection results are shown in Tables 1-1 to 1-3.
表1-1实施例1试剂的准确度分析结果Table 1-1 Accuracy analysis results of reagents in Example 1
表1-2实施例2试剂的准确度分析结果Table 1-2 Accuracy analysis results of reagents in Example 2
表1-3实施例3试剂的准确度分析结果Table 1-3 Accuracy analysis results of reagents in Example 3
结果显示，根据检测结果计算出的实施例1试剂检测方法的R2值为0.995，实施例2、3检测方法的R2值同样大于0.95，表明本发明方法检测结果与对比试剂盒检测结果无明显差异，具有较高准确度(符合度)。The results showed that the R2 value of the reagent detection method in Example 1 calculated according to the detection results was 0.995, and the R2 values of the detection methods in Examples 2 and 3 were also greater than 0.95, indicating that the detection results of the method of the present invention were not consistent with the detection results of the comparative kits. Significant difference, with high accuracy (conformity).
实施例5：本发明检测方法的精密度分析Example 5: Precision analysis of the detection method of the present invention
试验仪器：日立7080全自动生化分析仪；Test instrument: Hitachi 7080 automatic biochemical analyzer;
检测样品：低值、中值、高值血清样品各一份；中值血清样本一份；Test samples: one serum sample of low value, median value and high value; one serum sample of median value;
采用实施例1至实施例3的试剂及检测方法对同一待测样品重复检测10次，其检测结果见表2。The same sample to be tested was repeatedly detected 10 times using the reagents and detection methods of Examples 1 to 3, and the detection results are shown in Table 2.
表2检测样品脂蛋白(a)浓度(10次)及标准差率(变异系数)Table 2 Detection of sample lipoprotein (a) concentration (10 times) and standard deviation rate (coefficient of variation)
结果显示，实例1、2、3低值，中值，高值样本标准差率分别为2.72％、2.07％、2.27％；1.45％、1.07％、1.12％；1.19％、0.99％、1.12％，均小于3％，且小于对比试剂的标准差率，表明本发明方法具有高精密度。The results show that the sample standard deviation rates of low, median and high value samples in Examples 1, 2, and 3 are 2.72%, 2.07%, 2.27%; 1.45%, 1.07%, 1.12%; 1.19%, 0.99%, 1.12%, respectively. All are less than 3%, and less than the standard deviation rate of the comparative reagent, indicating that the method of the present invention has high precision.
实施例6：本发明方法的抗干扰性能分析Example 6: Anti-interference performance analysis of the method of the present invention
试验仪器：日立7080全自动生化分析仪；Test instrument: Hitachi 7080 automatic biochemical analyzer;
检测样品：任意一血清样品：Test sample: any serum sample:
在同一份血清样本中加入不同浓度的类风湿因子(50，150，250，500、800IU/mL)，采用实施例1至实施例3的检测方法对同一待测样品重复检测3次，与未加入类风湿因子的血清做对比，计算偏差，其检测结果见表3。在本实施例中，选取市售国产脂蛋白(a)试剂盒(胶乳增强免疫比浊法)作为对比试剂，进行了比较。Different concentrations of rheumatoid factor (50, 150, 250, 500, 800 IU/mL) were added to the same serum sample, and the same sample to be tested was tested 3 times using the detection methods of Example 1 to Example 3. The serum added with rheumatoid factor was compared, and the deviation was calculated. The test results are shown in Table 3. In this example, a commercially available domestic lipoprotein (a) kit (latex-enhanced immune turbidimetric method) was selected as a comparative reagent for comparison.
表3检测样品脂蛋白(a)浓度的平均值和偏差绝对值Table 3 The mean value and the absolute value of deviation of the lipoprotein (a) concentration of the detection samples
结果显示，本发明的试剂类风湿因子≤800IU/mL对测值没有影响，而对比试剂盒1仅在类风湿因子<300IU/mL对测值没有影响。The results show that the reagent rheumatoid factor of the present invention ≤ 800 IU/mL has no effect on the measured value, while the comparative kit 1 has no effect on the measured value only when the rheumatoid factor is less than 300 IU/mL.
以上实例中市售国产脂蛋白(a)试剂盒(胶乳增强免疫比浊法)的配方如下：The formulation of the commercially available domestic lipoprotein (a) kit (latex-enhanced immune turbidimetry) in the above example is as follows:
试剂1：40mmol/L含1.0％NaN3的甘氨酸缓冲液Reagent 1: 40mmol/L glycine buffer containing 1.0% NaN3
试剂2：抗人脂蛋白(a)-IgG的致敏乳胶颗粒悬液Reagent 2: Sensitized latex particle suspension of anti-human lipoprotein(a)-IgG
以上仅是本发明的优选实施方式，应当指出，对于本技术领域的普通技术人员来说，在不脱离本发明原理的前提下，还可以做出若干改进和润饰，这些改进和润饰也应视为本发明的保护范围。The above are only the preferred embodiments of the present invention. It should be pointed out that for those skilled in the art, some improvements and modifications can be made without departing from the principles of the present invention, and these improvements and modifications should also be regarded as It is the protection scope of the present invention.
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710399760.8ACN106990234B (en) | 2017-05-31 | 2017-05-31 | A kind of detection reagent and method of lipoprotein (a) |
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710399760.8ACN106990234B (en) | 2017-05-31 | 2017-05-31 | A kind of detection reagent and method of lipoprotein (a) |
| Publication Number | Publication Date |
|---|---|
| CN106990234A CN106990234A (en) | 2017-07-28 |
| CN106990234Btrue CN106990234B (en) | 2019-03-26 |
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201710399760.8AExpired - Fee RelatedCN106990234B (en) | 2017-05-31 | 2017-05-31 | A kind of detection reagent and method of lipoprotein (a) |
| Country | Link |
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| CN (1) | CN106990234B (en) |
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| CN103149370A (en)* | 2013-02-27 | 2013-06-12 | 宁波美康生物科技股份有限公司 | Lipoprotein (a) detection kit |
| CN103454193A (en)* | 2013-09-05 | 2013-12-18 | 苏州照康生物技术有限公司 | Immunoturbidimetric kit for detecting lipoprotein (a) and preparation method thereof |
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