CN108802375A - 一种抗核抗体谱的检测试剂盒 - Google Patents
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Abstract
本发明公开了一种抗核抗体谱的检测试剂盒,属于检测试剂盒技术领域。包括复合微珠悬浮液,所述复合微珠悬浮液中含有多个微珠,其中一部分微珠上分别包被有不同种类的自身抗原,一部分微珠上包被有从HEp‑2细胞中制备的抗原,还有一部分微珠上包被有一种能在病人样本中检测出非特异性抗体的抗原,剩余的微珠为四种标准对照微珠;荧光标记物,所述荧光标记物为藻红蛋白标记的羊抗‑人IgG(特异性γ链);样本稀释液,洗涤液。能够在同一时间同一孔内对一个病人样本做多个自身抗体的检测,从而对***性红斑狼疮(SLE)、类风湿性关节炎、硬皮症、多肌炎、混合型***疾病(MCTD)、药物诱导的SLE和干燥综合症等多种***性自身免疫疾病的提供辅助诊断。
Description
技术领域
本发明涉及一种抗核抗体谱的检测试剂盒,属于检测试剂盒技术领域。
背景技术
虽然目前自身免疫性疾病的确切病因不详,但在各种自身免疫***病患者中有特异性地自身抗体存在是确定的。对疑似各类***病患者,要联合和经常检测SSA52 kDa, SSA 60 kDa, SSB, Sm, U snRNP B/B’, U1 snRNP 68, U1 snRNP A, U1 snRNPC, Scl-70, Jo-1, Centromere B, PM Scl, Nucleosome, Ribosomal P, PCNA, dsDNA,Histone H, Histone HLY 和 M2等自身抗体,特别是这些IgG抗体。现有技术中,通过试剂盒检测时都是单独检测,效率低下。
发明内容
本发明所要解决的技术问题在于:提供一种抗核抗体谱的检测试剂盒,它解决了现有抗体检测效率低下的问题。
本发明所要解决的技术问题采取以下技术方案来实现:
一种抗核抗体谱的检测试剂盒,所述试剂盒包括,
复合微珠悬浮液,所述复合微珠悬浮液中含有多个微珠,其中一部分微珠上分别包被有不同种类的自身抗原,一部分微珠上包被有从HEp-2细胞中制备的抗原,还有一部分微珠上包被有一种能在病人样本中检测出非特异性抗体的抗原,剩余的微珠为四种标准对照微珠;
荧光标记物,所述荧光标记物为藻红蛋白标记的羊抗人IgG(特异性γ链);
样本稀释液,所述样本稀释液中含有磷酸盐缓冲液;
洗涤液,所述洗涤液中含有磷酸盐缓冲液。
作为优选实例,其中一分部微珠上分别包被有以下自身抗原:SSA 52 kDa, SSA60 kDa, SSB, Sm, U snRNP B/B’, U1 snRNP 68, U1 snRNP A, U1 snRNP C, Scl-70,Jo-1, Centromere B, PM Scl, Nucleosome, Ribosomal P, PCNA, dsDNA, Histone H,Histone HLY 和 M2。
作为优选实例,复合微珠悬浮液的组分以及其浓度为,
作为优选实例,复合微珠悬浮液中的Tween20用0.1-0.5g/L的十二烷基硫酸钠或0.5-1.5ml/L的曲拉通X-100代替。
作为优选实例, 复合微珠悬浮液中的Sodium Azide用0.5g /L-5g/L的硫柳汞代替。
作为优选实例, 藻红蛋白标记的羊抗人IgG(特异性γ链)的初始浓度为1.0 ug/mL,加入复合微珠悬浮液稀释定标至浓度0.6ug/L。
本发明的有益效果是:能够在同一时间同一孔内对一个病人样本做多个自身抗体的检测,从而对***性红斑狼疮(SLE),类风湿性关节炎,硬皮症(和***性硬皮病),多肌炎,混合型***疾病(MCTD),药物诱导的SLE和干燥综合症等多种***性自身免疫疾病的提供辅助诊断。本试剂盒用以辅助诊断多种***性自身免疫疾病,它能检测并区分多种抗核酸抗体和细胞质细胞组分,在一次检测中即可完成多种抗体的检测,极大提高了检测效率。
具体实施方式
为了对本发明的技术手段、创作特征、达成目的与功效易于明白了解,下面结合具体实施例,进一步阐述本发明。
一种抗核抗体谱的检测试剂盒,所述试剂盒包括,
复合微珠悬浮液,所述复合微珠悬浮液中含有多个微珠,其中一部分微珠上分别包被有不同种类的自身抗原,一部分微珠上包被有从HEp-2细胞中制备的抗原,还有一部分微珠上包被有一种能在病人样本中检测出非特异性抗体的抗原,剩余的微珠为四种标准对照微珠;
荧光标记物,所述荧光标记物为藻红蛋白标记的羊抗人IgG(特异性γ链);
样本稀释液,所述样本稀释液中含有磷酸盐缓冲液;
洗涤液,所述样本稀释液中含有磷酸盐缓冲液。
在其中的一种实施例中,其中一分部微珠上分别包被有以下自身抗原:SSA 52kDa, SSA 60 kDa, SSB, Sm, U snRNP B/B’, U1 snRNP 68, U1 snRNP A, U1 snRNP C,Scl-70, Jo-1, Centromere B, PM Scl, Nucleosome, Ribosomal P, PCNA, dsDNA,Histone H, Histone HLY 和 M2。
在其中的一种实施例中,复合微珠悬浮液的组分以及其浓度为,
在其中的一种实施例中,复合微珠悬浮液中的Tween20用0.1-0.5g/L的十二烷基硫酸钠或0.5-1.5ml/L的曲拉通X-100代替。
在其中的一种实施例中, 复合微珠悬浮液中的Sodium Azide用0.5g /L-5g/L的硫柳汞代替。
在其中的一种实施例中, 藻红蛋白标记的羊抗-人IgG(特异性γ链)的初始浓度为1.0 ug/ml,加入复合微珠悬浮液稀释定标至浓度0.6ug/L。
本试剂盒用以辅助诊断多种***性自身免疫疾病,它能检测并区分多种抗核酸抗体和细胞质细胞组分。以下表格列出了常见***性自身免疫疾病中自身抗体和疾病之间的关系。
制备过程:
1、制备复合微珠悬浮液
复合微珠悬浮液, 也用作样品稀释液、洗液。根据以下浓度配比计算需要的物质用量,配制试剂所需要的用量。
2、微珠的活化与包被
(1)取1ml特定编号微珠加入1.5ml离心管,加入5-50ul EDC和5-50ul Sulfo-NHS, 加入10-100ug 特定抗原(标准对照微珠1加入2-5ug/ml的人IgG,标准对照微珠2加入6-10ug/ml的人IgG,标准对照微珠3加入12-18ug/ml的人IgG,标准对照微珠4加入8-15ug/ml的抗人IgG,再采用NSC微珠作为空白对照,其不加入任何抗原),涡旋混匀30秒,超声混匀30秒,2-8°C摇匀过夜。
如果生产抗核抗体谱n项,需要同时放置n+5种抗原抗体(瓶子)的摇床,分别包被n+4种不同编号的微珠。
(2)洗涤:使用抽滤洗涤,经洗涤后的微珠转移至复合微珠悬浮液中。
(3)通过使用质控盘来验证质量。
(4)将多种已包被微珠混合,用少量复合微珠悬浮液洗下瓶壁残留微珠,并混合,最后用复合微珠悬浮液定容至120ml,检验合格后,分装成20瓶,每瓶5.5ml(100人份)。
3、荧光标记物:
藻红蛋白标记的羊抗人IgG(IgG-PE): 初始浓度1ug/ml,用复合微珠悬浮液稀释至工作浓度即可,工作浓度为0.6ug/L。
4、浓缩洗涤液:微珠保存液配方的10倍浓缩版,即本来配成1L的溶液,定容配成100ml。
上述微珠均为5.6微米的聚苯乙烯颗粒。
本发明能够在同一时间同一孔内对一个病人样本做多个自身抗体的检测,从而对***性红斑狼疮(SLE),类风湿性关节炎,硬皮症(和***性硬皮病),多肌炎,混合型***疾病(MCTD),药物诱导的SLE和干燥综合症等多种***性自身免疫疾病的提供辅助诊断。本试剂盒用以辅助诊断多种***性自身免疫疾病,它能检测并区分多种抗核酸抗体和细胞质细胞组分,在一次检测中即可完成多种抗原的检测,极大提高了检测效率。
以上显示和描述了本发明的基本原理和主要特征和本发明的优点。本行业的技术人员应该了解,本发明不受上述实施例的限制,在不脱离本发明精神和范围的前提下,本发明还会有各种变化和改进,这些变化和改进都落入本发明要求保护的范围内。本发明要求保护范围由所附的权利要求书及其等效物界定。
Claims (6)
1.一种抗核抗体谱的检测试剂盒,其特征在于:所述试剂盒包括,
复合微珠悬浮液,所述复合微珠悬浮液中含有多个微珠,其中一部分微珠上分别包被有不同种类的自身抗原,一部分微珠上包被有从HEp-2细胞中制备的抗原,还有一部分微珠上包被有一种能在病人样本中检测出非特异性抗体的抗原,剩余的微珠为四种标准对照微珠;
荧光标记物,所述荧光标记物为藻红蛋白标记的羊抗人IgG(特异性γ链);
样本稀释液,所述样本稀释液中含有磷酸盐缓冲液;
洗涤液,所述样本稀释液中含有磷酸盐缓冲液。
2.根据权利要求1所述的一种抗核抗体谱的检测试剂盒,其特征在于:其中一分部微珠上分别包被有以下自身抗原:SSA 52 kDa, SSA 60 kDa, SSB, Sm, U snRNP B/B’, U1snRNP 68, U1 snRNP A, U1 snRNP C, Scl-70, Jo-1, Centromere B, PM Scl,Nucleosome, Ribosomal P, PCNA, dsDNA, Histone H, Histone HLY 和 M2。
3.根据权利要求1所述的一种抗核抗体谱的检测试剂盒,其特征在于:复合微珠悬浮液的组分以及其浓度为,
。
4.根据权利要求2所述的一种抗核抗体谱的检测试剂盒,其特征在于:复合微珠悬浮液中的Tween20用0.1-0.5g/L的十二烷基硫酸钠或0.5-1.5ml/L的曲拉通X-100代替。
5.根据权利要求2所述的一种抗核抗体谱的检测试剂盒,其特征在于: 复合微珠悬浮液中的Sodium Azide用0.5g /L-5g/L的硫柳汞代替。
6.根据权利要求1所述的一种抗核抗体谱的检测试剂盒,其特征在于: 藻红蛋白标记的羊抗-人IgG(特异性γ链)的初始浓度为1.0 ug/mL,加入复合微珠悬浮液稀释定标至浓度0.6ug/L。
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CN113917161A (zh) * | 2021-10-18 | 2022-01-11 | 北京和杰创新生物医学科技有限公司 | 抗核周因子抗体免疫球蛋白g的检测方法 |
CN114814203A (zh) * | 2022-04-20 | 2022-07-29 | 北京胡曼智造科技有限责任公司 | 一种抗核抗体联合检测试剂盒 |
CN114814203B (zh) * | 2022-04-20 | 2023-09-15 | 北京胡曼智造科技有限责任公司 | 一种抗核抗体联合检测试剂盒 |
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