[1]宋姝庭,孙 丽,蒋金凤,等.基于实时荧光定量监测逆转录病毒MuLV/Friend感染方法的建立与评价[J].中国药理学通报,2018,(08):1175-1178.[doi:10.3969/j.issn.1001-1978.2018.08.027]
 SONG Shu-ting,SUN Li,JIANG Jin-feng,et al.Establishment and validation of real-time quantitative reverse transcription PCR assay for detecting replication dynamics of MuLV/Friend in mice[J].Chinese Pharmacological Bulletin,2018,(08):1175-1178.[doi:10.3969/j.issn.1001-1978.2018.08.027]
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基于实时荧光定量监测逆转录病毒MuLV/Friend感染方法的建立与评价()
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《中国药理学通报》[ISSN:/CN:]

卷:
期数:
2018年08期
页码:
1175-1178
栏目:
实验方法学
出版日期:
2018-08-26

文章信息/Info

Title:
Establishment and validation of real-time quantitative reverse transcription PCR assay for detecting replication dynamics of MuLV/Friend in mice
文章编号:
1001-1978(2018)08-1175-04
作者:
宋姝庭12孙 丽2蒋金凤2黄 海1王建华2
1.上海大学生命科学学院,上海 200444; 2.中国科学院上海巴斯德研究所,上海 200031
Author(s):
SONG Shu-ting12SUN Li2JIANG Jin-feng2HUANG Hai1WANG Jian-hua2
1.School of Life Sciences,Shanghai University,Shanghai 200444,China; 2.Institute Pasteur of Shanghai,Chinese Academy of Sciences,Shanghai 200031,China
关键词:
MuLV/Friend病毒 逆转录病毒 急性感染 病毒复制 实时荧光核酸定量 动物模型
Keywords:
MuLV/Friend virus retrovirus acute infection viral replication real-time quantitative reverse transcription PCR assay animal model
分类号:
R-332; R363-332; R373.9; R342.2; R446.9; R512.9; R733.7
DOI:
10.3969/j.issn.1001-1978.2018.08.027
文献标志码:
A
摘要:
目的 建立基于实时荧光定量PCR的病毒复制检测方法,监测MuLV/Friend急性感染小鼠病毒复制动力学。方法 提取MuLV/Friend病毒基因组RNA,逆转录为cDNA,扩增病毒gag片段,连接入PMD-19T载体,构建标准品,并合成特异性荧光探针,建立病毒复制定量的标准曲线; BALB/c小鼠腹腔接种MuLV/Friend病毒,在急性感染不同时期,实时监测小鼠组织内病毒复制动力学。结果 成功构建基于实时荧光定量PCR的MuLV/Friend复制检测方法,可分析小鼠各组织内病毒复制动力学,为利用MuLV/Friend小鼠感染模型研究抗病毒策略或宿主免疫等,提供了一种有效、快速、灵敏的实时监测病毒复制的手段。结论 该实时荧光定量PCR的病毒复制检测方法具有较高的灵敏度,改善了之前利用脾肿大指数检测病毒体内复制的传统方法。
Abstract:
Aim To establish the real-time quantitative reverse transcriptase PCR(qRT-PCR)-based approach for quantifying the replication dynamics of MuLV/Friend in vivo.Methods The genome RNA from MuLV/Friend was extracted and reverse transcribed into cDNA; gag DNA was amplified and then linked into PMD-19T vector to build a standard substance,the specific fluorescent probe against gag was synthesized,and the standard curve of gag DNA was established by qRT-PCR quantification; BALB/c mice were infected via intraperitoneal injection with MuLV/Friend,and the viral replication dynamics in multiple tissues was quantified by qRT-PCR.Results The qRT-PCR approach was established to quantify MuLV/Friend replication in mice.The approach was effective,faster and sensitive to monitor the dynamics of viral replication in multiple tissues of infected mice,and would facilitate the use of the MuLV/Friend-infected mouse model in the studies of host antiviral immune responses and the development of antiviral strategies.Conclusion The established qRT-PCR approach in this study has higher sensitivity for detecting viral particles,which may improve the traditional method for detecting viral replication by calculating the index of splenomegaly.

参考文献/References:

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备注/Memo

备注/Memo:
收稿日期:2018-04-25,修回日期:2018-05-28
基金项目:国家自然科学基金青年基金资助项目(No 81601750)
作者简介:宋姝庭(1991-),女,硕士生,研究方向:病毒免疫学,E-mail:stsong@ips.ac.cn;
王建华(1977-),男,博士,研究员,博士生导师,研究方向:病毒免疫学,通讯作者,Tel:021-54923120,E-mail:jh_wang@sibs.ac.cn
更新日期/Last Update: 2018-07-26