(RT)-qPCR用于hiv -1慢病毒载体RNA和DNA的检测和分化。

Q1 Immunology and Microbiology

Human Gene Therapy Methods Pub Date : 2017-07-27 DOI:10.1089/hgtb.2017.081

M. Pavlovic, Nina Koehler, M. Anton, Anna Dinkelmeier, Maren Haase, Thorsten Stellberger, U. Busch, A. Baiker

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引用次数: 2

摘要

所述方法的目的是检测和区分细胞培养上清液和拭子样本中hiv衍生慢病毒载体(LV)的RNA和DNA。对于基因工程操作的分析监测，需要检测基于HIV-1的LV世代的方法。此外，在研究问题上，证明LV颗粒的不存在对于降低实验环境的生物安全水平非常重要。本文描述了一种针对LTR U5亚基和封装信号ψ起始序列的qPCR方法。为了监控技术过程，包括了许多控制措施。

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(RT)-qPCR for detection of and differentiation between RNA and DNA of HIV-1-based lentiviral vectors.

The purpose of the described method is the detection of and differentiation between RNA and DNA of HIV-derived lentiviral vectors (LV) in cell culture supernatants and swab samples. For the analytical surveillance of genetic engineering operations methods for the detection of the HIV-1 based LV generations are required. Furthermore, for research issues, it is important to prove the absence of LV particles for downgrading experimental settings in terms of the biosafety level. Here a qPCR method targeting the LTR U5 subunit and the start sequence of the packaging signal ψ is described. Numerous controls are included in order to monitor the technical procedure.

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来源期刊

Human Gene Therapy Methods BIOTECHNOLOGY & APPLIED MICROBIOLOGY-GENETICS & HEREDITY

CiteScore

5.80

自引率

0.00%

发文量

审稿时长

>12 weeks

期刊介绍： Human Gene Therapy is the premier, multidisciplinary journal covering all aspects of gene therapy. The Journal publishes in-depth coverage of DNA, RNA, and cell therapies by delivering the latest breakthroughs in research and technologies. Human Gene Therapy provides a central forum for scientific and clinical information, including ethical, legal, regulatory, social, and commercial issues, which enables the advancement and progress of therapeutic procedures leading to improved patient outcomes, and ultimately, to curing diseases. The Journal is divided into three parts. Human Gene Therapy, the flagship, is published 12 times per year. HGT Methods, a bimonthly journal, focuses on the applications of gene therapy to product testing and development. HGT Clinical Development, a quarterly journal, serves as a venue for publishing data relevant to the regulatory review and commercial development of cell and gene therapy products.