CN108330184A - 一种快速筛选特应性皮炎样本的方法 - Google Patents

一种快速筛选特应性皮炎样本的方法 Download PDF

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CN108330184A
CN108330184A CN201710038447.1A CN201710038447A CN108330184A CN 108330184 A CN108330184 A CN 108330184A CN 201710038447 A CN201710038447 A CN 201710038447A CN 108330184 A CN108330184 A CN 108330184A
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sample
atopic dermatitis
mirna
quick screening
standard items
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陈小帆
曹恒昌
邹彦芬
杨旸
张李娟
于波
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SHENZHEN PKU-HKUST MEDICAL CENTER
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SHENZHEN PKU-HKUST MEDICAL CENTER
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    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
    • C12Q1/6844—Nucleic acid amplification reactions
    • C12Q1/686—Polymerase chain reaction [PCR]

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Abstract

本发明公开了一种快速筛选特应性皮炎样本的方法。本发明方法是通过使用一个标准品miRNA‑151a，与血清样本中提取的RNA一同进行特异性的逆转录，得到的逆转录产物在进行荧光定量PCR以检测样品和标准品中miRNA‑151a浓度的差异，从而确定样本是否为特应性皮炎样本，此方法较传统方法快速准确，能高效、大通量的分辨出特应性皮炎样本。

Description

一种快速筛选特应性皮炎样本的方法
技术领域
本发明涉及分子生物学领域，具体涉及一种快速筛选特应性皮炎样本的方法。
背景技术
特应性皮炎（atopic dermatitis，AD）是一种慢性炎症性皮肤病，在全世界范围内儿童发病率高达15%-30%，成人发病率约为2%-10%。特应性皮炎呈现一系列广谱的临床特征，包括初期出现红斑、干燥发痒乃至后期出现红皮性牛皮癣等严重的症状。部分特应性皮炎患者可并发其他过敏性疾病，同时因为病情反复、睡眠缺失、饮食限制以及其他社会因素使患者的生活质量下降。
实验室对特应性皮炎的研究经常需要用到特应性皮炎患者的血液，但收集的患者血液经常会与荨麻疹或其他皮肤病患者的血液混在一起，因而需要区分出特应性皮炎患者的血液，以进行下一步的研究。但样本患者的信息通常并不齐全。现有实验室的技术，仅能通过IgE浓度用间接手段粗略筛选特应性皮炎患者，准确度不高，且耗时较长，急需一种高效精准的筛选特应性皮炎患者血液的方法。
发明内容
本发明的目的在于提供一种快速筛选特应性皮炎样本的方法。
本发明上述目的通过以下技术方案予以实现：
通过提取患者血液中的RNA，进行特异性的逆转录，而后进行荧光定量即可筛选出特应性皮炎患者。
与现有技术相比，本发明具有如下有益效果：
本发明将使得实验室特应性皮炎患者血液的筛选变得精准快速。
附图说明
图1为实时荧光定量PCR检测样本及标准品的miRNA-151a的吸光检测图；
图2为特应性皮炎患者血清样品、标准品和正常人血清样品用实时荧光定量PCR检测miRNA-151a结果的散点图。
具体实施方式
以下结合实施例来进一步解释本发明，但实施例并不对本发明做任何形式的限定。
1. 取200ul样品血清，使用Trizol提取RNA。
2. 取合成miRNA-151a样品，配置成1pg/mL的样品，取200ul作为标准品。
3. 使用miRNA-151a特异性逆转录引物（SEQ ID NO:1）逆转录miRNA-151a。
4. 使用PCR引物扩增miRNA-151a（SEQ ID NO:2、SEQ ID NO:3）。
5. 扩增结果如图1所示，取CT值做散点图得图2，CT值小于标准品则为特应性皮炎患者血清样本，CT值大于标准品则为正常人血清样本。
申请人：深圳北京大学香港科技大学医学中心
发明名称：一种快速筛选特应性皮炎样本的方法
SEQ ID NO:1
名称：miRNA-151a-3p-RT
序列：GTCGTATCCAGTGCGTGTCGTGGAGTCGGCAATTGCACTGGATACGACCCTCAAG
SEQ ID NO:2
名称：miR151a-3p-F
序列：GCCACTAGACTGAAGCTC
SEQ ID NO:3
名称：mirR
序列：GTGCGTGTCGTGGAGTC

Claims (1)

1.一种快速筛选特应性皮炎样本的方法，其特征在于使用一个合成miRNA-151a作为标准品，将其与血清样本提出的RNA一起进行特异性逆转录，而后在通过荧光定量与之相比较，得出结论样本是否为特应性皮炎样本。
CN201710038447.1A 2017-01-19 2017-01-19 一种快速筛选特应性皮炎样本的方法 Pending CN108330184A (zh)

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Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN112094895A (zh) * 2020-08-18 2020-12-18 河北仁博科技有限公司 特应性皮炎诊断和治疗的miRNA
CN113604569A (zh) * 2021-08-04 2021-11-05 广州医科大学 miR-6883-3p在制备用于抗肝癌或肝癌预后评估产品中的应用

Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN103173448A (zh) * 2013-02-07 2013-06-26 上海市胸科医院 早期肺腺癌miRNA特异表达谱及其反转录引物和用途
WO2014075822A1 (en) * 2012-11-16 2014-05-22 Siemens Aktiengesellschaft New diagnostic mirna markers for parkinson disease
CN104181307A (zh) * 2014-08-21 2014-12-03 深圳北京大学香港科技大学医学中心 一种通过ctack/ccl27评估特异性皮炎病情的诊断方法
CN105039530A (zh) * 2015-07-03 2015-11-11 南京医科大学 线粒体相关精浆微小核糖核酸作为人类严重弱精子症发生的标志物及其应用

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2014075822A1 (en) * 2012-11-16 2014-05-22 Siemens Aktiengesellschaft New diagnostic mirna markers for parkinson disease
CN103173448A (zh) * 2013-02-07 2013-06-26 上海市胸科医院 早期肺腺癌miRNA特异表达谱及其反转录引物和用途
CN104181307A (zh) * 2014-08-21 2014-12-03 深圳北京大学香港科技大学医学中心 一种通过ctack/ccl27评估特异性皮炎病情的诊断方法
CN105039530A (zh) * 2015-07-03 2015-11-11 南京医科大学 线粒体相关精浆微小核糖核酸作为人类严重弱精子症发生的标志物及其应用

Non-Patent Citations (4)

* Cited by examiner, † Cited by third party
Title
XIAOFAN CHEN等: ""MiR‐151a is involved in the pathogenesis of atopic dermatitis by regulating interleukin‐12 receptor β2"", 《EXPERIMENTAL DERMATOLOGY》 *
于建华等: "《实用临床基础检验》", 31 August 2014, 西安交通大学出版社 *
张李娟等: ""特应性皮炎患者血清microRNA的差异性表达"", 《中华实用诊断与治疗杂志》 *
陈超等: "《生物技术检验检疫实践教程》", 31 March 2016, 华南理工大学出版社 *

Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN112094895A (zh) * 2020-08-18 2020-12-18 河北仁博科技有限公司 特应性皮炎诊断和治疗的miRNA
CN112094895B (zh) * 2020-08-18 2021-08-13 河北仁博科技有限公司 特应性皮炎诊断和治疗的miRNA
CN113604569A (zh) * 2021-08-04 2021-11-05 广州医科大学 miR-6883-3p在制备用于抗肝癌或肝癌预后评估产品中的应用
CN113604569B (zh) * 2021-08-04 2023-11-17 广州医科大学 miR-6883-3p在制备用于抗肝癌或肝癌预后评估产品中的应用

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