CN111413500A - 一种新型冠状病毒2019-nCoV抗原胶体金检测技术 - Google Patents
一种新型冠状病毒2019-nCoV抗原胶体金检测技术 Download PDFInfo
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- G—PHYSICS
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- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/58—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving labelled substances
- G01N33/583—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving labelled substances with non-fluorescent dye label
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- 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
- G01N33/558—Immunoassay; Biospecific binding assay; Materials therefor using diffusion or migration of antigen or antibody
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- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
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- 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
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- G01N2469/10—Detection of antigens from microorganism in sample from host
Abstract
本专利属于医疗体外诊断领域，涉及一种新型冠状病毒2019‑nCoV抗原胶体金检测技术，步骤包括样品制备、加样层析、质量控制和结果分析与判定。通过观察质控线C和检测线T的颜色变化判断新型冠状病毒抗原阴性和阳性情况。本专利检测周期短且操作简便，可在新冠肺炎疑似病例确诊、解除隔离和出院标准判断过程中发挥重要作用。
Description
技术领域
本专利属于医疗体外诊断领域，涉及一种新型冠状病毒2019-nCoV抗原胶体金检测技 术，尤其是涉及一种检测周期短且操作简便的新型冠状病毒2019-nCoV抗原胶体金检测技 术。
背景技术
在2019-nCoV的防控过程中，快速检测试剂发挥了非常重要的作用。快速诊断试剂的 灵敏性、特异性和稳定性对于2019-nCoV病例的早期发现、早期隔离和早期治疗具有非常 重要的意义，也是目前控制病毒传染源最可靠的方法。快速诊断分抗原检测和抗体检测两类， 抗原检测方法包括检测病毒表面抗原蛋白和检测特异核酸片段，用于确认体内是否存在病原 体，适用于疾病潜伏期、急性期或病程初期。抗体检测一般用于判断病程进展程度或用于判 断是否发生既往感染。兼具特异性、灵敏度、易操作、时间短等优势的聚合酶链式反应(P CR)、胶体金或荧光层析等技术是临床体外诊断常用的方法。
免疫胶体金检测技术具有更高检测效率，可以在10-15分钟内得到结果。免疫胶体金技 术(Immune colloidal gold technique)是以胶体金作为示踪标志物应用于抗原抗体的一种新 型的免疫标记技术，根据标记的示踪标志物的不同，标记抗原以检测抗体，标记抗体以检测 抗原，可以分别实现对抗体和病原体的检测。该技术可实现抗原的快速即时检验(Point-of- care Testing，POCT)，在采样现场即刻进行分析，省去了样品运输时间和实验室复杂处理 程序，通常无需专业的实验室场地、人员和设备，即便需要特定的试剂和仪器也为便携式的。 同时，具有检测时间短、操作便捷、肉眼即可判读结果等优势，特别适合疫情严重时的现场 快速筛查。一旦该方法可成熟应用于新型冠状病毒的检测，不仅适于医院使用，更可以在基 层医疗机构和家庭使用，具有极高的临床使用价值。除应用于临床病人的筛选和确诊之外， 对于研究新型冠状病毒编码蛋白质的抗原性、抗体产生特点及规律、疫苗的评价等也具有重 要意义。
因此，开发一种检测周期短且操作简便的新型冠状病毒2019-nCoV抗原胶体金检测技 术具有重要意义。
发明内容
本专利的有益之处在于，可以提供一种检测周期短且操作简便的新型冠状病毒2019-n CoV抗原胶体金检测技术。
为实现上述功能，本专利涉及的一种新型冠状病毒2019-nCoV抗原胶体金检测技术， 其特征在于，步骤如下：
1)样品制备
在生物安全柜内，取0.5ml样本稀释液，将咽拭子浸泡到样本稀释液中充分搅拌，并 将咽拭子中的液体在管壁挤干净，盖上管盖，充分混匀，室温放置5min待测；
2)加样层析
取出检测卡，平放于水平桌面上，做好标记，用滴管或移液器从盛有带侧样品的试管 中取2-3滴或100μl样品滴加于样品孔内，于15min内观察测试结果；
3)质量控制
可见质控线C呈显紫红色条带；
4)结果分析与判定
在检测有效的情况下，检测线T呈现紫红色条，可判断为新型冠状病毒抗原阳性；T线不显色，可判断为新型冠状病毒抗原阴性。
其中，样本稀释液为0.02mol/L磷酸氢二钠和磷酸二氢钠盐缓冲液，检测线为预包被 0.2μg鼠抗2019-nCoV，金标记物为金颗粒标记的0.1μg兔抗2019-nCoV，质控线为预包被0.1μg羊抗兔IgG。
具体实施方式
为详细说明本专利的技术内容、所实现目的及效果，以下结合实施例予以说明：
实施例一
取咽拭子样本按步骤检测。
1)样品制备
在生物安全柜内，取0.5ml样本稀释液，将咽拭子浸泡到样本稀释液中充分搅拌，并 将咽拭子中的液体在管壁挤干净，盖上管盖，充分混匀，室温放置5min待测；
2)加样层析
取出检测卡，平放于水平桌面上，做好标记，用滴管或移液器从盛有带侧样品的试管 中取2-3滴或100μl样品滴加于样品孔内，于15min内观察测试结果；
经检测，样本提取液滴加入样品孔内后，质控线C呈显紫红色条带，T线不显色，说明检测有效，且样本为新冠状病毒抗原阴性。
上述实施例仅供说明本专利之用，而并非是对本专利的限制；应当指出的是，对于本 领域的普通技术人员，在不脱离本专利构思范围的情况下，还可以作出各种变化和变型， 这些都属于本专利的保护范围；因此，凡跟本专利权利要求范围所做的均等变化与修饰， 均应属于本专利权利要求的覆盖范围。
Claims (2)
1.一种新型冠状病毒2019-nCoV抗原胶体金检测技术，其特征在于，步骤如下：
1)样品制备
在生物安全柜内，取0.5ml样本稀释液，将咽拭子浸泡到样本稀释液中充分搅拌，并将咽拭子中的液体在管壁挤干净，盖上管盖，充分混匀，室温放置5min待测；
2)加样层析
取出检测卡，平放于水平桌面上，做好标记，用滴管或移液器从盛有带侧样品的试管中取2-3滴或100μl样品滴加于样品孔内，于15min内观察测试结果；
3)质量控制
可见质控线C呈显紫红色条带；
4)结果分析与判定
在检测有效的情况下，检测线T呈现紫红色条，可判断为新型冠状病毒抗原阳性；T线不显色，可判断为新型冠状病毒抗原阴性。
2.根据权利要求1所述的一种新型冠状病毒2019-nCoV抗原胶体金检测技术，其特征在于，样本稀释液为0.02mol/L磷酸氢二钠和磷酸二氢钠盐缓冲液，检测线为预包被0.2μg鼠抗2019-nCoV，金标记物为金颗粒标记的0.1μg兔抗2019-nCoV，质控线为预包被0.1μg羊抗兔IgG。
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114456259A (zh) * | 2020-11-07 | 2022-05-10 | 上海科技大学 | 一种抗新冠病毒的抗体及其制备方法和应用 |
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| CN1453589A (zh) * | 2003-06-18 | 2003-11-05 | 广州万孚生物技术有限公司 | 胶体金层析法检测sars冠状病毒抗体的试剂 |
| CN1570638A (zh) * | 2003-07-22 | 2005-01-26 | 李克生 | Sars病毒抗体检测方法及快速诊断试剂盒和制备方法 |
| KR101782862B1 (ko) * | 2016-04-26 | 2017-09-29 | 원광대학교산학협력단 | 메르스바이러스 진단용 단세포군 항체 및 면역크로마토그래피 진단키트 |
| CN111286558A (zh) * | 2020-02-12 | 2020-06-16 | 谱尼测试集团北京检验认证科学研究院有限公司 | 一种新型冠状病毒2019-nCoV核酸检测技术 |
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- 2020-02-12 CN CN202010096343.8A patent/CN111413500A/zh active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1453589A (zh) * | 2003-06-18 | 2003-11-05 | 广州万孚生物技术有限公司 | 胶体金层析法检测sars冠状病毒抗体的试剂 |
| CN1570638A (zh) * | 2003-07-22 | 2005-01-26 | 李克生 | Sars病毒抗体检测方法及快速诊断试剂盒和制备方法 |
| KR101782862B1 (ko) * | 2016-04-26 | 2017-09-29 | 원광대학교산학협력단 | 메르스바이러스 진단용 단세포군 항체 및 면역크로마토그래피 진단키트 |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN114456259A (zh) * | 2020-11-07 | 2022-05-10 | 上海科技大学 | 一种抗新冠病毒的抗体及其制备方法和应用 |
| CN114456259B (zh) * | 2020-11-07 | 2023-06-02 | 上海科技大学 | 一种抗新冠病毒的抗体及其制备方法和应用 |
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