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
- Publication number
- CN107727866A CN107727866A CN201710914326.9A CN201710914326A CN107727866A CN 107727866 A CN107727866 A CN 107727866A CN 201710914326 A CN201710914326 A CN 201710914326A CN 107727866 A CN107727866 A CN 107727866A
- Authority
- CN
- China
- Prior art keywords
- reagent
- preparation
- latex
- detection kit
- dispersion liquid
- 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.)
- Withdrawn
Links
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
- G01N33/6893—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids related to diseases not provided for elsewhere
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/543—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals
- G01N33/54313—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals the carrier being characterised by its particulate form
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/543—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals
- G01N33/544—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals the carrier being organic
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
- G01N33/6887—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids from muscle, cartilage or connective tissue
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
- G01N2333/435—Assays involving biological materials from specific organisms or of a specific nature from animals; from humans
- G01N2333/46—Assays involving biological materials from specific organisms or of a specific nature from animals; from humans from vertebrates
- G01N2333/47—Assays involving proteins of known structure or function as defined in the subgroups
- G01N2333/4701—Details
- G01N2333/4712—Muscle proteins, e.g. myosin, actin, protein
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2800/00—Detection or diagnosis of diseases
- G01N2800/32—Cardiovascular disorders
- G01N2800/325—Heart failure or cardiac arrest, e.g. cardiomyopathy, congestive heart failure
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。
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN201710914326.9A CN107727866A (zh) | 2017-09-30 | 2017-09-30 | 一种n‑端脑利钠肽前体检测试剂盒的制备方法 |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN201710914326.9A CN107727866A (zh) | 2017-09-30 | 2017-09-30 | 一种n‑端脑利钠肽前体检测试剂盒的制备方法 |
Publications (1)
Publication Number | Publication Date |
---|---|
CN107727866A true CN107727866A (zh) | 2018-02-23 |
Family
ID=61209473
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN201710914326.9A Withdrawn CN107727866A (zh) | 2017-09-30 | 2017-09-30 | 一种n‑端脑利钠肽前体检测试剂盒的制备方法 |
Country Status (1)
Country | Link |
---|---|
CN (1) | CN107727866A (zh) |
Cited By (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN111141914A (zh) * | 2020-01-02 | 2020-05-12 | 四川纳海川生物科技有限公司 | 一种n-端脑利肽前体检测试剂盒及其制备方法 |
Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN102161716A (zh) * | 2010-12-30 | 2011-08-24 | 北京九强生物技术股份有限公司 | 胶乳致敏的方法及试剂 |
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 |
-
2017
- 2017-09-30 CN CN201710914326.9A patent/CN107727866A/zh not_active Withdrawn
Patent Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN102161716A (zh) * | 2010-12-30 | 2011-08-24 | 北京九强生物技术股份有限公司 | 胶乳致敏的方法及试剂 |
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 |
Cited By (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN111141914A (zh) * | 2020-01-02 | 2020-05-12 | 四川纳海川生物科技有限公司 | 一种n-端脑利肽前体检测试剂盒及其制备方法 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
CN105486859B (zh) | 一种新型改进免疫层析试纸条及其制备和应用 | |
WO2018120620A1 (zh) | 荧光免疫层析检测卡及其制备方法和应用 | |
CN104697988B (zh) | 检测丙型肝炎病毒抗体的试剂盒及其检测方法和应用 | |
WO2016127318A1 (zh) | 心肌肌钙蛋白i超敏检测试剂盒及其超敏检测方法 | |
Chandler et al. | A new enzyme immunoassay system suitable for field use and its application in a snake venom detection kit | |
CN101988926A (zh) | 一种高尔基蛋白gp73的夹心elisa定量检测方法及其检测试剂盒 | |
CN101661040A (zh) | 梅毒螺旋体抗体唾液快速检测试纸条 | |
JP4790818B2 (ja) | 上昇した量のd−ダイマーを含む保存安定性細胞性全血組成物 | |
CN111868528A (zh) | 侧向流动免疫测定条状装置 | |
CN106546750A (zh) | 一种微量白蛋白快速定量检测试纸条的制备及检测方法 | |
JP4976068B2 (ja) | 簡易イムノアッセイ用検体浮遊液およびアッセイ方法 | |
CN114814214A (zh) | 一种胶体金、乳胶微球标记联合的星状病毒免疫层析检测试剂盒及制备方法 | |
CN107727866A (zh) | 一种n‑端脑利钠肽前体检测试剂盒的制备方法 | |
Villalta et al. | Autoantibodies to asialoglycoprotein receptor (ASGPR) in patients with autoimmune liver diseases | |
CN107655879A (zh) | 用于检测性腺系列的磁微粒的微流控化学发光检测系统 | |
CN110702926A (zh) | 一种胃泌素g17检测试剂盒及其制备方法 | |
CN202141725U (zh) | 一种定性/定量检测乙型肝炎病毒表面抗原的磁珠免疫层析试剂盒 | |
CN103645321A (zh) | 一种筛查降钙素原的试纸及其制备方法 | |
CN107748254A (zh) | 一种n‑端脑利钠肽前体检测试剂盒及其应用 | |
CN109030816A (zh) | 用于诊断肝素诱导血小板减少症的活化试验 | |
CN202837303U (zh) | Ngal荧光免疫层析定量检测试纸条 | |
Bosch et al. | Analysis of urinary human growth hormone (hGH) using hydrogel nanoparticles and isoform differential immunoassays after short recombinant hGH treatment: preliminary results | |
CN107741492A (zh) | 一种包被有n‑端脑利钠肽前体抗体的胶乳微球的制备方法 | |
CN106932593B (zh) | 以ncam‑1为检测指标的双抗体夹心酶联免疫吸附检测试剂盒及其应用 | |
JP2006118936A (ja) | メンブランエンザイムイムノアッセイ法 |
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 | ||
WW01 | Invention patent application withdrawn after publication | ||
WW01 | Invention patent application withdrawn after publication |
Application publication date: 20180223 |