CN106814063B - 一种稳定的血清铁检测试剂盒 - Google Patents
一种稳定的血清铁检测试剂盒 Download PDFInfo
- Publication number
- CN106814063B CN106814063B CN201510859182.2A CN201510859182A CN106814063B CN 106814063 B CN106814063 B CN 106814063B CN 201510859182 A CN201510859182 A CN 201510859182A CN 106814063 B CN106814063 B CN 106814063B
- Authority
- CN
- China
- Prior art keywords
- reagent
- iron
- serum levels
- detection kit
- kit
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
- 238000001514 detection method Methods 0.000 title claims abstract description 30
- 210000002966 serum Anatomy 0.000 title claims abstract description 30
- XEEYBQQBJWHFJM-UHFFFAOYSA-N Iron Chemical compound [Fe] XEEYBQQBJWHFJM-UHFFFAOYSA-N 0.000 title claims description 119
- 229910052742 iron Inorganic materials 0.000 title claims description 60
- 239000003153 chemical reaction reagent Substances 0.000 claims abstract description 71
- 238000000034 method Methods 0.000 claims abstract description 16
- 239000007983 Tris buffer Substances 0.000 claims abstract description 12
- LENZDBCJOHFCAS-UHFFFAOYSA-N tris Chemical compound OCC(N)(CO)CO LENZDBCJOHFCAS-UHFFFAOYSA-N 0.000 claims abstract description 12
- 239000000872 buffer Substances 0.000 claims abstract description 4
- CWYNVVGOOAEACU-UHFFFAOYSA-N Fe2+ Chemical compound [Fe+2] CWYNVVGOOAEACU-UHFFFAOYSA-N 0.000 claims description 15
- GLUUGHFHXGJENI-UHFFFAOYSA-N diethylenediamine Natural products C1CNCCN1 GLUUGHFHXGJENI-UHFFFAOYSA-N 0.000 claims description 12
- PXIPVTKHYLBLMZ-UHFFFAOYSA-N Sodium azide Chemical compound [Na+].[N-]=[N+]=[N-] PXIPVTKHYLBLMZ-UHFFFAOYSA-N 0.000 claims description 10
- CIWBSHSKHKDKBQ-JLAZNSOCSA-N Ascorbic acid Chemical compound OC[C@H](O)[C@H]1OC(=O)C(O)=C1O CIWBSHSKHKDKBQ-JLAZNSOCSA-N 0.000 claims description 6
- 239000000203 mixture Substances 0.000 claims description 4
- 229960005070 ascorbic acid Drugs 0.000 claims description 3
- 239000002211 L-ascorbic acid Substances 0.000 claims description 2
- 235000000069 L-ascorbic acid Nutrition 0.000 claims description 2
- 239000004094 surface-active agent Substances 0.000 abstract description 6
- 230000005293 ferrimagnetic effect Effects 0.000 abstract description 3
- QMPKHBWKQLDRGR-UHFFFAOYSA-N iron;piperazine Chemical compound [Fe].C1CNCCN1 QMPKHBWKQLDRGR-UHFFFAOYSA-N 0.000 abstract description 3
- 238000012360 testing method Methods 0.000 description 9
- 208000007502 anemia Diseases 0.000 description 6
- 210000004185 liver Anatomy 0.000 description 6
- 102000008857 Ferritin Human genes 0.000 description 5
- 108050000784 Ferritin Proteins 0.000 description 5
- 238000008416 Ferritin Methods 0.000 description 5
- 239000003795 chemical substances by application Substances 0.000 description 5
- VTLYFUHAOXGGBS-UHFFFAOYSA-N Fe3+ Chemical compound [Fe+3] VTLYFUHAOXGGBS-UHFFFAOYSA-N 0.000 description 4
- 238000002835 absorbance Methods 0.000 description 4
- 229910001448 ferrous ion Inorganic materials 0.000 description 4
- 210000002540 macrophage Anatomy 0.000 description 4
- -1 oxygen radical Chemical class 0.000 description 4
- 102000001554 Hemoglobins Human genes 0.000 description 3
- 108010054147 Hemoglobins Proteins 0.000 description 3
- 108010017480 Hemosiderin Proteins 0.000 description 3
- 206010028851 Necrosis Diseases 0.000 description 3
- 108010015865 Transferrins Proteins 0.000 description 3
- 102000002070 Transferrins Human genes 0.000 description 3
- 210000004027 cell Anatomy 0.000 description 3
- 230000008859 change Effects 0.000 description 3
- 238000006243 chemical reaction Methods 0.000 description 3
- 208000019425 cirrhosis of liver Diseases 0.000 description 3
- 238000004737 colorimetric analysis Methods 0.000 description 3
- 201000010099 disease Diseases 0.000 description 3
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 3
- 238000002474 experimental method Methods 0.000 description 3
- 239000007789 gas Substances 0.000 description 3
- JEIPFZHSYJVQDO-UHFFFAOYSA-N iron(III) oxide Inorganic materials O=[Fe]O[Fe]=O JEIPFZHSYJVQDO-UHFFFAOYSA-N 0.000 description 3
- 210000005229 liver cell Anatomy 0.000 description 3
- 238000005259 measurement Methods 0.000 description 3
- 230000017074 necrotic cell death Effects 0.000 description 3
- 230000009972 noncorrosive effect Effects 0.000 description 3
- 210000000056 organ Anatomy 0.000 description 3
- 239000002504 physiological saline solution Substances 0.000 description 3
- 102000004190 Enzymes Human genes 0.000 description 2
- 108090000790 Enzymes Proteins 0.000 description 2
- 206010016654 Fibrosis Diseases 0.000 description 2
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 2
- 208000036142 Viral infection Diseases 0.000 description 2
- 239000002253 acid Substances 0.000 description 2
- 210000004369 blood Anatomy 0.000 description 2
- 239000008280 blood Substances 0.000 description 2
- 230000019522 cellular metabolic process Effects 0.000 description 2
- 230000001684 chronic effect Effects 0.000 description 2
- 230000007882 cirrhosis Effects 0.000 description 2
- 150000001875 compounds Chemical class 0.000 description 2
- 238000003745 diagnosis Methods 0.000 description 2
- 230000029142 excretion Effects 0.000 description 2
- 230000012010 growth Effects 0.000 description 2
- 239000001301 oxygen Substances 0.000 description 2
- 229910052760 oxygen Inorganic materials 0.000 description 2
- 210000000952 spleen Anatomy 0.000 description 2
- 102000007592 Apolipoproteins Human genes 0.000 description 1
- 108010071619 Apolipoproteins Proteins 0.000 description 1
- 208000037157 Azotemia Diseases 0.000 description 1
- 241000894006 Bacteria Species 0.000 description 1
- 230000004543 DNA replication Effects 0.000 description 1
- 206010019837 Hepatocellular injury Diseases 0.000 description 1
- DGAQECJNVWCQMB-PUAWFVPOSA-M Ilexoside XXIX Chemical compound C[C@@H]1CC[C@@]2(CC[C@@]3(C(=CC[C@H]4[C@]3(CC[C@@H]5[C@@]4(CC[C@@H](C5(C)C)OS(=O)(=O)[O-])C)C)[C@@H]2[C@]1(C)O)C)C(=O)O[C@H]6[C@@H]([C@H]([C@@H]([C@H](O6)CO)O)O)O.[Na+] DGAQECJNVWCQMB-PUAWFVPOSA-M 0.000 description 1
- 206010061218 Inflammation Diseases 0.000 description 1
- 206010022971 Iron Deficiencies Diseases 0.000 description 1
- 206010065973 Iron Overload Diseases 0.000 description 1
- 206010022979 Iron excess Diseases 0.000 description 1
- 206010028980 Neoplasm Diseases 0.000 description 1
- 206010029164 Nephrotic syndrome Diseases 0.000 description 1
- TZRXHJWUDPFEEY-UHFFFAOYSA-N Pentaerythritol Tetranitrate Chemical compound [O-][N+](=O)OCC(CO[N+]([O-])=O)(CO[N+]([O-])=O)CO[N+]([O-])=O TZRXHJWUDPFEEY-UHFFFAOYSA-N 0.000 description 1
- 208000037581 Persistent Infection Diseases 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
- 230000003213 activating effect Effects 0.000 description 1
- 230000032683 aging Effects 0.000 description 1
- 210000001188 articular cartilage Anatomy 0.000 description 1
- 235000010323 ascorbic acid Nutrition 0.000 description 1
- 239000011668 ascorbic acid Substances 0.000 description 1
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 210000000988 bone and bone Anatomy 0.000 description 1
- 239000007853 buffer solution Substances 0.000 description 1
- 239000012928 buffer substance Substances 0.000 description 1
- 230000003139 buffering effect Effects 0.000 description 1
- 230000030833 cell death Effects 0.000 description 1
- 210000000170 cell membrane Anatomy 0.000 description 1
- 230000004663 cell proliferation Effects 0.000 description 1
- 238000005336 cracking Methods 0.000 description 1
- 230000001351 cycling effect Effects 0.000 description 1
- 230000006378 damage Effects 0.000 description 1
- 230000037213 diet Effects 0.000 description 1
- 235000005911 diet Nutrition 0.000 description 1
- 230000004069 differentiation Effects 0.000 description 1
- 238000010790 dilution Methods 0.000 description 1
- 239000012895 dilution Substances 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 230000002124 endocrine Effects 0.000 description 1
- 230000012202 endocytosis Effects 0.000 description 1
- 230000002708 enhancing effect Effects 0.000 description 1
- 210000003743 erythrocyte Anatomy 0.000 description 1
- 229910001447 ferric ion Inorganic materials 0.000 description 1
- 238000000705 flame atomic absorption spectrometry Methods 0.000 description 1
- 239000012530 fluid Substances 0.000 description 1
- 208000007475 hemolytic anemia Diseases 0.000 description 1
- 230000002440 hepatic effect Effects 0.000 description 1
- 206010073071 hepatocellular carcinoma Diseases 0.000 description 1
- 231100000844 hepatocellular carcinoma Toxicity 0.000 description 1
- 231100000437 hepatocellular injury Toxicity 0.000 description 1
- 231100000086 high toxicity Toxicity 0.000 description 1
- 108010045676 holotransferrin Proteins 0.000 description 1
- 230000004054 inflammatory process Effects 0.000 description 1
- 230000031891 intestinal absorption Effects 0.000 description 1
- 210000004347 intestinal mucosa Anatomy 0.000 description 1
- 230000003834 intracellular effect Effects 0.000 description 1
- 230000037427 ion transport Effects 0.000 description 1
- 230000010438 iron metabolism Effects 0.000 description 1
- 210000003734 kidney Anatomy 0.000 description 1
- 230000002045 lasting effect Effects 0.000 description 1
- 230000003859 lipid peroxidation Effects 0.000 description 1
- 201000007270 liver cancer Diseases 0.000 description 1
- 208000019423 liver disease Diseases 0.000 description 1
- 208000014018 liver neoplasm Diseases 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 230000004060 metabolic process Effects 0.000 description 1
- 230000003680 myocardial damage Effects 0.000 description 1
- 210000004165 myocardium Anatomy 0.000 description 1
- 102000039446 nucleic acids Human genes 0.000 description 1
- 108020004707 nucleic acids Proteins 0.000 description 1
- 150000007523 nucleic acids Chemical class 0.000 description 1
- 230000004768 organ dysfunction Effects 0.000 description 1
- 210000000496 pancreas Anatomy 0.000 description 1
- 210000004738 parenchymal cell Anatomy 0.000 description 1
- 230000007918 pathogenicity Effects 0.000 description 1
- 230000001766 physiological effect Effects 0.000 description 1
- 230000035935 pregnancy Effects 0.000 description 1
- 230000001737 promoting effect Effects 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 238000003908 quality control method Methods 0.000 description 1
- 230000009467 reduction Effects 0.000 description 1
- 230000035945 sensitivity Effects 0.000 description 1
- 229910052708 sodium Inorganic materials 0.000 description 1
- 239000011734 sodium Substances 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 210000004243 sweat Anatomy 0.000 description 1
- 238000003786 synthesis reaction Methods 0.000 description 1
- 231100000331 toxic Toxicity 0.000 description 1
- 230000002588 toxic effect Effects 0.000 description 1
- 231100000419 toxicity Toxicity 0.000 description 1
- 230000001988 toxicity Effects 0.000 description 1
- 239000011573 trace mineral Substances 0.000 description 1
- 235000013619 trace mineral Nutrition 0.000 description 1
- 230000032258 transport Effects 0.000 description 1
- 208000009852 uremia Diseases 0.000 description 1
- 210000002700 urine Anatomy 0.000 description 1
- 238000010200 validation analysis Methods 0.000 description 1
- 238000012795 verification Methods 0.000 description 1
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/75—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated
- G01N21/77—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated by observing the effect on a chemical indicator
- G01N21/78—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated by observing the effect on a chemical indicator producing a change of colour
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/17—Systems in which incident light is modified in accordance with the properties of the material investigated
- G01N21/25—Colour; Spectral properties, i.e. comparison of effect of material on the light at two or more different wavelengths or wavelength bands
- G01N21/31—Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry
Landscapes
- Physics & Mathematics (AREA)
- Chemical & Material Sciences (AREA)
- General Health & Medical Sciences (AREA)
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- General Physics & Mathematics (AREA)
- Immunology (AREA)
- Pathology (AREA)
- Spectroscopy & Molecular Physics (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Plasma & Fusion (AREA)
- Engineering & Computer Science (AREA)
- Investigating Or Analysing Biological Materials (AREA)
Abstract
本发明公开了一种血清铁亚铁嗪法检测试剂盒，属于临床体外检测试剂技术领域。本发明试剂盒包括试剂R1和试剂R2。通过将试剂R1与试剂R2的缓冲液更换为60mM pH为4.2的Tris缓冲液，同时将试剂R1的表面活性剂更换为TritonX‑100，提高了试剂盒的稳定性，线性相关性较好，试剂的准确度也较好，有利于在市场中进一步的推广使用。
Description
技术领域
本发明涉及临床体外检测试剂技术领域，特别涉及一种血清铁免疫比浊法检测试剂盒。
背景技术
铁是人体内的主要微量元素之一，常以三价铁离子的形式与转铁蛋白结合并进行运输。铁在细胞代谢的过程中具有重要作用，适量的铁在生物机体氧转运、细胞生物氧化、细菌致病性、氧自由基的控制、细胞中DNA复制及细胞繁殖等方面都有重要作用。此外，铁离子还可以参与血红蛋白、肌红蛋白以及人体必需酶的合成，在人体代谢及各种生理活动过程中具有至关重要的作用。
正常情况下，人体每天由尿液、汗液等排出的铁与吸收的铁含量保持平衡。在正常生理状态下，约5%的铁由肠道吸收来满足人体的正常生理需求，约95%的铁来源于巨噬细胞，红细胞衰老裂解后被巨噬细胞吞噬，其解离出的铁离子泵回血液再利用。人体的铁主要分为两大部分，第一部分是功能铁，含量为35~45mg/kg，主要是血红蛋白，还有少量的肌红蛋白及细胞代谢、生长、分化所需的含铁酶。另一部分为储存铁，主要分布在肝脏、脾脏及骨髓的单核巨噬细胞，以铁蛋白及含铁血黄素的形式存在。
人体内储存铁的含量波动较大，为0~20mg/kg，超过此值则为铁过载。过多的铁沉积在肝脏、脾脏、胰腺、骨髓、心肌、肾脏、肠粘膜、关节软骨及内分泌器官等部位，其中肝脏和心肌损伤可能出现严重的临床并发症。人体内铁缺乏可引起缺铁性贫血等疾病，但铁过多也会对人体产生毒害作用，引起肝炎、肝细胞坏死等疾病。肝脏是铁储存的主要器官，尤其是肝实质细胞中含量最多，铁含量过多时肝脏是最主要的受损器官。研究表明，铁超载致使肝细胞损伤加重，同时促进肝细胞癌及肝纤维化的发生。
机体的铁代谢是相对封闭的，主要是循环再利用。人体对铁的排泄是持续的、强制性的、有一定限度的，不会因为铁的超负荷而增加铁的排泄。巨噬细胞释放铁主要是以转铁蛋白的形式，其次还存在有毒性的非转铁蛋白结合铁的形式。细胞外与铁蛋白结合的铁会通过细胞内吞作用进入细胞内的不稳定铁库，此种形式的铁有很高的毒性。细胞内暂时无用的铁与载脂蛋白结合进行储存，这种铁蛋白是没有毒性的。当这种铁蛋白的含量超过不稳定铁库容量时，铁蛋白就会变性，形成含铁血黄素，这种形式的铁毒性更高。不稳定铁库中有二价铁，也有三价铁，二价铁可以与脂质过氧化物反应，产生羟基自由基和脂质过氧化物，损害细胞膜、蛋白质和核酸，进而致使细胞死亡及器官功能障碍。含铁血黄素的慢性沉积，引发肝细胞坏死、肝硬化，少数还会发展为肝癌。
血清铁是临床上常测的微量元素之一，是鉴别诊断贫血，辅助诊断肝病和肿瘤的指标，其测定在临床上愈来愈受到人们的重视。在临床上，血清铁降低见于缺铁饮食、吸收不良、慢性失血、妊娠，或婴幼儿生长发育需铁量增高所致缺铁性贫血及慢性感染、肝硬化、尿毒症、肾病综合症。血清铁升高见于铁剂治疗过量、溶血性贫血、再生障碍性贫血、巨幼细胞贫血、血红蛋白生成障碍性贫血及急性肝炎、肝细胞坏死等。
血清铁的检测方法有：亚铁嗪法比色法、火焰原子吸收光谱法光法、干化学法等，这些方法在灵敏度、特异度、准确性方面都有很大差异。在酸性条件下，血清铁从与其结合的转铁蛋白上游离出来，同时三价铁被还原为亚铁离子（二价铁），亚铁离子与亚铁嗪反应形成紫色复合物。在570 nm处测其吸光度变化，与标本中的铁离子浓度成正比。测定血清中的铁对临床许多疾病的诊断和疗效观察都有非常重要的临床意义。
鉴于此，本发明在亚铁嗪比色法的基础上，优化反应体系，选用Tris缓冲液及表面活性剂TritonX-100，显著改善了试剂的稳定性，发现了一种更稳定的血清铁检测试剂。
发明内容
本发明的目的是提供一种稳定的用于检测血清铁的试剂盒，该试剂盒采用亚铁嗪比色法。该试剂盒与常规的试剂盒相比，稳定性和线性相关性比常规的检测试剂盒要好，有利于试剂在临床上的推广应用。
基本原理：
在酸性条件下，血清铁从与其结合的转铁蛋白上游离出来。同时，三价铁被还原为亚铁离子（二价铁），亚铁离子与亚铁嗪反应形成紫色复合物。在570 nm处测得的吸光度变化，与标本中的铁离子浓度成正比。
本发明是通过以下步骤得到的：
一种血清铁检测试剂盒，它包含试剂R1和试剂R2。其特征在于，试剂组成如下：试剂R1为60mmol/L pH 4.2的Tris缓冲液、100mmol/L抗坏血酸、10mmol/L叠氮钠、1%TritonX-100；试剂R2为60mmol/L pH 4.2的Tris缓冲液、50mmol/L亚铁嗪、10mmol/L叠氮钠。
所述的血清铁检测试剂盒，其特征在于所述表面活性剂为TritonX-100。
所述的血清铁检测试剂来检测血清铁的检测方法，使用全自动生化分析仪利用终点法进行测定，检测主波长为570nm。
所述的血清铁检测试剂，试剂R1和试剂R2在使用时的比例为R1:R2=200: 50。
本发明的试剂盒在具有双试剂功能的全自动生化分析仪上进行，其具体使用方法见附图3：
加入生理盐水、样本或校准品20μl，再加入200μl 的R1试剂预孵育5min后读取吸光度A1，之后再加入100μl的R2试剂反应5min后，读取吸光度A2，并计算ΔA。
本发明的有益效果：
1）试剂R1和试剂R2采用新的缓冲体系，改善了试剂的稳定性；
2）试剂R1采用新的表面活性剂TritonX-100，增强了试剂的稳定性，而且不会对试剂的准确度产生影响；
3）试剂的准确度和稳定性良好，价格便宜，使用方便，有利于该试剂在市场中进一步的推广。
附图说明
图1为两种试剂的相关性曲线图；
图2为两种试剂效期稳定性曲线图；
图3为本发明试剂盒使用方法流程图，
图中R1:试剂1，R2:试剂2，S:校准品，U:样本。
具体实施方式
下面结合具体实施例对本发明进行进一步说明：
实施例1
一种常规的血清铁检测试剂盒，它包括试剂R1和试剂R2。
其中试剂R1组成为：
pH 4.2 Tris缓冲液 60mmol/L
抗坏血酸 100mmol/L
叠氮钠 10mmol/L
表面活性剂TritonX-100 1%
试剂R2组成为：
pH 4.2 Tris缓冲液 60mmol/L
亚铁嗪 50mmol/L
叠氮钠 10mmol/L
本实施例所述试剂R1和试剂R2，配置时需先加入缓冲物质Tris，用盐酸调到适当pH值后，再添加其它物质。本实施例所述试剂盒，在使用时，其测定方法是采用具有双试剂功能的迈瑞800全自动生化分析仪，利用终点法进行测定，操作如下：
加入生理盐水、样本或校准品20μl，再加入200μl 的R1试剂预孵育5min后读取吸光度A1，之后再加入100μl的R2试剂反应5min后，读取吸光度A2，并计算ΔA。
ΔA＝A2- A1
血清铁含量（μmol/L）＝（∆A测定÷∆A标准）×C标准
测定：检测样本 ∆A标准：标准品
实施例2
准确度验证试验：将实施例1的血清铁检测试剂作为实验组，市场上获得认可的一种准确度好、稳定性好的血清铁检测试剂盒作为对照组进行检测，对20个临床血清样本进行检测，检测结果如表1所示。获得了两种试剂的相关性曲线（如图1所示），结果表明，两组试剂盒的相关系数为0.9989，说明两者相关性比较好。证明本发明试剂盒添加及更改的组分对其准确性不会造成影响，试剂盒依然保持较好的准确度。
表1实施例1试剂与市场认可的血清铁检测试剂盒对比检测结果
实施例3
线性相关性验证试验：配制铁含量为180μmol/L的高值样本，用生理盐水进行系列稀释，配制6个不同浓度的样本，浓度依次为180μmol/L、144μmol/L、108μmol/L、72μmol/L、36μmol/L、0μmol/L。分别利用实施例1试剂及对照组试剂进行检测，每个浓度的样本分别测定三次，分别取平均值，检测结果如表2所示。
表2 实施例1试剂线性相关验证实验检测结果
如上表所示，实施例1与对照组试剂检测结果相关系数均大于0.990，且实施例1试剂检测结果的相关系数略大于对照组试剂检测结果的相关系数，这表明本发明试剂具有更好的线性相关性。
实施例4
稳定性验证实验：在2℃～8℃、无腐蚀性气体的避光环境中贮存试剂，检测实施例1及对照组试剂的稳定性。两组试剂每月选取同一样本测定质控品（靶值19.2μmol/L），测定三次取平均值，检测数据如表3所示。
表3 实施例1试剂稳定性验证实验检测结果
实验结果显示，实施例1试剂在2℃～8℃、无腐蚀性气体的避光环境中贮存15个月稳定，而对照组试剂在2℃～8℃、无腐蚀性气体的避光环境中贮存10个月后开始不稳定，说明本发明试剂更换缓冲液及表面活性剂后稳定性增强。
综上所述，本发明提供的血清铁亚铁嗪法检测试剂盒，试剂R1与试剂R2中的缓冲液更换为60 mmol/L pH为4.2的Tris缓冲液中，同时将试剂R1中的表面活性剂更换为TritonX-100 ，提高了试剂盒的稳定性，线性相关性好，试剂的准确度也较好。因此，本发明提供的血清铁亚铁嗪法检测试剂盒有利于在市场中进一步的推广使用。
Claims (4)
1.一种血清铁检测试剂盒，其特征在于包含试剂R1和试剂R2；试剂组成如下：试剂R1组成为：60mmol/LpH 4.2的Tris缓冲液、100mmol/L抗坏血酸、10mmol/L叠氮钠、1％TritonX-100；试剂R2组成为：60mmol/LpH 4.2的Tris缓冲液、50mmol/L亚铁嗪、10mmol/L叠氮钠；
所述试剂R1和试剂R2使用时的比例为R1:R2＝200:50。
2.根据权利要求1所述的血清铁检测试剂盒，其特征在于试剂R1缓冲液为25℃，pH为4.2的Tris缓冲液。
3.根据权利要求1所述的血清铁检测试剂盒，其特征在于试剂R2缓冲液为25℃，pH为4.2的Tris缓冲液。
4.根据权利要求1所述的血清铁检测试剂盒，其特征在于，使用全自动生化分析仪采用终点法进行测定，检测主波长为570nm。
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510859182.2A CN106814063B (zh) | 2015-12-01 | 2015-12-01 | 一种稳定的血清铁检测试剂盒 |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510859182.2A CN106814063B (zh) | 2015-12-01 | 2015-12-01 | 一种稳定的血清铁检测试剂盒 |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN106814063A CN106814063A (zh) | 2017-06-09 |
| CN106814063B true CN106814063B (zh) | 2019-03-12 |
Family
ID=59156374
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201510859182.2A Active CN106814063B (zh) | 2015-12-01 | 2015-12-01 | 一种稳定的血清铁检测试剂盒 |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN106814063B (zh) |
Families Citing this family (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109613280B (zh) * | 2018-12-29 | 2022-01-28 | 中拓生物有限公司 | 一种血清铁测定试剂盒及其制备方法和应用 |
| CN110865075A (zh) * | 2019-12-31 | 2020-03-06 | 南通大学 | 一种常规的尿铁检测试剂盒及其检测方法 |
Family Cites Families (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101629960A (zh) * | 2008-07-18 | 2010-01-20 | 中生北控生物科技股份有限公司 | 铁测定试剂组合、试剂盒及其应用 |
| CN104483494B (zh) * | 2014-12-22 | 2016-09-28 | 宁波美康生物科技股份有限公司 | 一种血清不饱和铁结合力检测试剂盒 |
-
2015
- 2015-12-01 CN CN201510859182.2A patent/CN106814063B/zh active Active
Also Published As
| Publication number | Publication date |
|---|---|
| CN106814063A (zh) | 2017-06-09 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Biswas et al. | Mechanism of erythrocyte death in human population exposed to arsenic through drinking water | |
| CN105264384B (zh) | 参比材料中不稳定分析物的稳定化 | |
| Ōyanagui | Reevaluation of assay methods and establishment of kit for superoxide dismutase activity | |
| Hastka et al. | Washing erythrocytes to remove interferents in measurements of zinc protoporphyrin by front-face hematofluorometry | |
| ES2804205T3 (es) | Método de análisis para componente de muestra biológica diluida | |
| CN103760357B (zh) | 一种缺血修饰白蛋白检测试剂盒 | |
| CN101458257B (zh) | 检测缺血修饰白蛋白的试剂盒及其制备方法 | |
| CN108627510A (zh) | 高密度脂蛋白胆固醇检测试剂盒 | |
| CN107153043A (zh) | 一种液体即用型抗凝血酶ⅲ活性测定试剂 | |
| CN103364519B (zh) | 黄嘌呤氧化酶抑制剂和/或超氧阴离子清除剂的筛选方法 | |
| Dent et al. | Measuring magnesium–Physiological, clinical and analytical perspectives | |
| CN106814063B (zh) | 一种稳定的血清铁检测试剂盒 | |
| CN112501245A (zh) | 一种新型N-乙酰-β-D氨基葡萄糖苷酶检测试剂 | |
| Warner et al. | Prognostic role of antioxidant enzymes in sepsis: preliminary assessment | |
| Fiorito et al. | Assessment of iron absorption in mice by ICP-MS measurements of 57 Fe levels | |
| Akram et al. | Development and validation of an economical uric acid-Fe3+/Fe2+-ferrozine-based colorimetric assay to estimate uric acid level of pure and biological samples | |
| Abusoglu et al. | Assessment of serum ischemia-modified albumin, prolidase and thiol-disulphide levels in subjects with breast cancer | |
| CN106814191A (zh) | 一种视黄醇结合蛋白免疫比浊法检测试剂盒 | |
| Reddy et al. | Gender differences in alcohol-induced oxidative stress and altered membrane properties in erythrocytes of rats | |
| Laursen et al. | The association between content of the elements S, Cl, K, Fe, Cu, Zn and Br in normal and cirrhotic liver tissue from Danes and Greenlandic Inuit examined by dual hierarchical clustering analysis | |
| CN106053851A (zh) | 稳定性高的前白蛋白检测试剂盒 | |
| Medzihradsky et al. | A simple procedure for the separation of viable blood cells, suitable for long-term in vitro experiments | |
| CN103674938A (zh) | 缺血修饰白蛋白组合测定试剂、测定方法及试剂盒 | |
| CN105181692A (zh) | 一种缺血修饰白蛋白比色法检测试剂盒 | |
| CN110865075A (zh) | 一种常规的尿铁检测试剂盒及其检测方法 |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant | ||
| PE01 | Entry into force of the registration of the contract for pledge of patent right | ||
| PE01 | Entry into force of the registration of the contract for pledge of patent right |
Denomination of invention: A Stable Serum Iron Detection Kit Effective date of registration: 20230628 Granted publication date: 20190312 Pledgee: Bank of Communications Ltd. Shandong branch Pledgor: BIOBASE BIODUSTRY (SHANDONG) Co.,Ltd. Registration number: Y2023980045843 |
|
| PC01 | Cancellation of the registration of the contract for pledge of patent right | ||
| PC01 | Cancellation of the registration of the contract for pledge of patent right |
Granted publication date: 20190312 Pledgee: Bank of Communications Ltd. Shandong branch Pledgor: BIOBASE BIODUSTRY (SHANDONG) Co.,Ltd. Registration number: Y2023980045843 |