CN107727866A - 一种n‑端脑利钠肽前体检测试剂盒的制备方法 - Google Patents
一种n‑端脑利钠肽前体检测试剂盒的制备方法 Download PDFInfo
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Abstract
本发明公开了一种N‑端脑利钠肽前体检测试剂盒的制备方法，包括试剂R1的制备和试剂R2的制备；其中，试剂R1的制备包括：用牛血清白蛋白、缓冲液中、PEG‑6000、蔗糖、海藻糖、防腐剂制得试剂R1；试剂R2的制备包括：用牛血清白蛋白、缓冲液、表面活性剂、防腐剂、胶乳微球、聚乙烯吡咯烷酮、N‑端脑利钠肽前体抗体，制得试剂R2。本发明的优点在于：该方法简单、快速、成本低、特异性强、稳定性好、重复性好。
Description
技术领域
本发明涉及医学检验技术领域，更具体涉及一种N-端脑利钠肽前体检测试剂盒的制备方法。
背景技术
心血管疾病是目前危害人类健康的重要疾病之一，中国每年因该疾病入院病人在三千多万，死于该疾病的有三百多万人，而且每年呈上升趋势。所以，利用合适的医疗设备对心血管疾病的诊断、治疗和康复已成为一个引起全球性重视的话题。N-端脑利钠肽前体(NT-ProBNP)是B型利钠肽激素原分裂后无活性的N端碎片，是典型的心脏标志物，主要在心肌细胞所受容量负荷增高时由左心室分泌。N-端脑利钠肽前体的应用在于协助诊断充血性心力衰竭，判断病情的严重程度和预后，以及指导治疗已经取得了医学界的广泛关注。
迄今，已有多篇相关的研究论文发表于新英格兰医学杂志、循环医学、美国心脏病学院杂志等权威杂志上。这些论文探讨了诸如NT-ProBNP在心衰急诊诊断中的作用、快速NT-ProBNP检测用于区别充血性心衰或肺部疾患导致的呼吸困难的病人等问题，为心衰的诊断和评估开辟了一个崭新的领域。
目前市面上检测NT-ProBNP的试剂盒主要是以荧光免疫分析法、酶联免疫分析法的方式进行检测，这些方法成本高、准确度低。还需要配备特定的仪器进行配合检测，操作比较繁琐。并且试剂盒的制备工艺复杂。
发明内容
本发明所要解决的技术问题在于提供了一种制备工艺简单的一种N-端脑利钠肽前体检测试剂盒的制备方法。
本发明是通过以下技术方案解决上述技术问题的：
一种N-端脑利钠肽前体检测试剂盒的制备方法，包括试剂R1的制备和试剂R2的制备；
其中，
试剂R1中各组分的含量为：
所述试剂R1的制备包括以下步骤：
(a1)将牛血清白蛋白溶于缓冲液中，搅拌均匀，待充分溶解，制得分散液；
(a2)将PEG-6000、蔗糖、海藻糖、防腐剂溶于步骤(a1)所制得的分散液中，搅拌均匀，待充分溶解，制得试剂R1。
试剂R2中各组分的含量为：
所述试剂R2的制备包括如下步骤：
(b1)将牛血清白蛋白溶于缓冲液中，搅拌均匀，待充分溶解，制得分散液；
(b2)将表面活性剂、防腐剂、胶乳微球、聚乙烯吡咯烷酮溶于步骤(b1)所制得的分散液中，搅拌均匀，制得乳胶微球溶解试剂；
(b3)采用化学偶联法将N-端脑利钠肽前体抗体包被到乳胶微球中，制得抗体胶乳；
(b4)将步骤(b3)制得的抗体胶乳离心去上清液，然后用步骤(b2)制得的乳胶微球溶解试剂溶解沉淀物，使抗体胶乳的终浓度为1.0％，超声分散，制得试剂R2。
优选地，所述缓冲液选自HEPES缓冲液或硼酸缓冲液。
优选地，所述防腐剂选自HY-500防腐剂或叠氮钠。
优选地，所述表面活性剂选自S9或曲拉通-100。
优选地，所述胶乳微球的粒径为80nm。
本发明相比现有技术具有以下优点：该方法简单、快速、成本低、特异性强、稳定性好、重复性好。
具体实施方式
下面对本发明的实施例作详细说明，本实施例在以本发明技术方案为前提下进行实施，给出了详细的实施方式和具体的操作过程，但本发明的保护范围不限于下述的实施例。
所有原料均为市购原料；所有配比中，对于固体，％表示g/100mL，对于液体，％表示mL/100mL。
实施例1
一种N-端脑利钠肽前体检测试剂盒的制备方法，包括试剂R1的制备和试剂R2的制备。
所有配比中，对于固体，％表示g/100mL，对于液体，％表示mL/100mL。
试剂R1的组分及含量为：
试剂R1的制备包括以下步骤：
(a1)将牛血清白蛋白溶于HEPES缓冲液中，搅拌均匀，待充分溶解，制得分散液；
(a2)将PEG-6000、蔗糖、海藻糖、HY-500防腐剂溶于步骤(a1)所制得的分散液中，搅拌均匀，待充分溶解，制得试剂R1。
试剂R2的组分及含量为：
试剂R2的制备包括如下步骤：
(b1)将牛血清白蛋白溶于HEPES缓冲液中，搅拌均匀，待充分溶解，制得分散液；
(b2)将表面活性剂S9、HY-500防腐剂、粒径为80nm的胶乳微球、聚乙烯吡咯烷酮溶于步骤(b1)所制得的分散液中，搅拌均匀，制得乳胶微球溶解试剂；
(b3)采用化学偶联法将N-端脑利钠肽前体抗体包被到乳胶微球中，制得抗体胶乳；
(b4)将步骤(b3)制得的抗体胶乳离心去上清液，然后用步骤(b2)制得的乳胶微球溶解试剂溶解沉淀物，使抗体胶乳的终浓度为1.0％，超声分散，制得试剂R2。
实施例2
一种N-端脑利钠肽前体检测试剂盒的制备方法，包括试剂R1的制备和试剂R2的制备。
试剂R1的制备包括以下步骤：
(a1)将牛血清白蛋白溶于硼酸缓冲液中，搅拌均匀，待充分溶解，制得分散液；
(a2)将PEG-6000、蔗糖、海藻糖、防腐剂叠氮钠溶于步骤(a1)所制得的分散液中，搅拌均匀，待充分溶解，制得试剂R1。
所述试剂R2的组分及含量为：
试剂R2的制备包括如下步骤：
(b1)将牛血清白蛋白溶于硼酸缓冲液中，搅拌均匀，待充分溶解，制得分散液；
(b2)将表面活性剂曲拉通-100、防腐剂叠氮钠、粒径为80nm的胶乳微球、聚乙烯吡咯烷酮溶于步骤(b1)所制得的分散液中，搅拌均匀，制得乳胶微球溶解试剂；
(b3)采用化学偶联法将N-端脑利钠肽前体抗体包被到乳胶微球中，制得抗体胶乳；
(b4)将步骤(b3)制得的抗体胶乳离心去上清液，然后用步骤(b2)制得的乳胶微球溶解试剂溶解沉淀物，使抗体胶乳的终浓度为1.0％，超声分散，制得试剂R2。
实施例3
一种N-端脑利钠肽前体检测试剂盒的制备方法，包括试剂R1的制备和试剂R2的制备。
试剂R1的组分及含量为：
试剂R1的制备包括以下步骤：
(a1)将牛血清白蛋白溶于HEPES缓冲液中，搅拌均匀，待充分溶解，制得分散液；
(a2)将PEG-6000、蔗糖、海藻糖、防腐剂叠氮钠溶于步骤(a1)所制得的分散液中，搅拌均匀，待充分溶解，制得试剂R1。
试剂R2的组分及含量为：
试剂R2的制备包括如下步骤：
(b1)将牛血清白蛋白溶于HEPES缓冲液中，搅拌均匀，待充分溶解，制得分散液；
(b2)将表面活性剂S9、防腐剂叠氮钠、粒径为80nm的胶乳微球、聚乙烯吡咯烷酮溶于步骤(b1)所制得的分散液中，搅拌均匀，制得乳胶微球溶解试剂；
(b3)采用化学偶联法将N-端脑利钠肽前体抗体包被到乳胶微球中，制得抗体胶乳；
(b4)将步骤(b3)制得的抗体胶乳离心去上清液，然后用步骤(b2)制得的乳胶微球溶解试剂溶解沉淀物，使抗体胶乳的终浓度为1.0％，超声分散，制得试剂R2。
该方法简单、快速、成本低、特异性强、稳定性好、重复性好。
以上所述仅为本发明的较佳实施例而已，并不用以限制本发明，凡在本发明的精神和原则之内所作的任何修改、等同替换和改进等，均应包含在本发明的保护范围之内。
Claims (5)
1.一种N-端脑利钠肽前体检测试剂盒的制备方法，其特征在于，包括试剂R1的制备和试剂R2的制备；
其中，试剂R1中各组分的含量为：
所述试剂R1的制备包括以下步骤：
(a1)将牛血清白蛋白溶于缓冲液中，搅拌均匀，待充分溶解，制得分散液；
(a2)将PEG-6000、蔗糖、海藻糖、防腐剂溶于步骤(a1)所制得的分散液中，搅拌均匀，待充分溶解，制得试剂R1；
试剂R2中各组分的含量为：
所述试剂R2的制备包括如下步骤：
(b1)将牛血清白蛋白溶于缓冲液中，搅拌均匀，待充分溶解，制得分散液；
(b2)将表面活性剂、防腐剂、胶乳微球、聚乙烯吡咯烷酮溶于步骤(b1)所制得的分散液中，搅拌均匀，制得乳胶微球溶解试剂；
(b3)采用化学偶联法将N-端脑利钠肽前体抗体包被到乳胶微球中，制得抗体胶乳；
(b4)将步骤(b3)制得的抗体胶乳离心去上清液，然后用步骤(b2)制得的乳胶微球溶解试剂溶解沉淀物，使抗体胶乳的终浓度为1.0％，超声分散，制得试剂R2。
2.根据权利要求1所述的N-端脑利钠肽前体检测试剂盒的制备方法，其特征在于，所述缓冲液选自HEPES缓冲液或硼酸缓冲液。
3.根据权利要求1所述的N-端脑利钠肽前体检测试剂盒的制备方法，其特征在于，所述防腐剂选自HY-500防腐剂或叠氮钠。
4.根据权利要求1所述的N-端脑利钠肽前体检测试剂盒的制备方法，其特征在于，所述表面活性剂选自S9或曲拉通-100。
5.根据权利要求1所述的N-端脑利钠肽前体检测试剂盒的制备方法，其特征在于，所述胶乳微球的粒径为80nm。
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| CN105431728A (zh) * | 2013-05-31 | 2016-03-23 | 积水医疗株式会社 | 凝集免疫测定法 |
| CN106093407A (zh) * | 2016-05-31 | 2016-11-09 | 安徽伊普诺康生物技术股份有限公司 | 一种测定脂蛋白(a)的试剂盒及其制备方法 |
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| CN105431728A (zh) * | 2013-05-31 | 2016-03-23 | 积水医疗株式会社 | 凝集免疫测定法 |
| CN106093407A (zh) * | 2016-05-31 | 2016-11-09 | 安徽伊普诺康生物技术股份有限公司 | 一种测定脂蛋白(a)的试剂盒及其制备方法 |
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