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Iranian Biomedical Journal، جلد ۲۲، شماره ۶، صفحات ۳۹۴-۴۰۰
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چکیده فارسی مقاله
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عنوان انگلیسی Preparation and Evaluation of Ribonuclease-Resistant Viral HIV RNA Standards Based on Armored RNA Technology
چکیده انگلیسی مقاله Background: The human immunodeficiency virus type 1 (HIV-1) is an infectious viral agent that gradually extinguishes the immune system, resulting in acquired immune deficiency syndrome (AIDS). The aim of this study was to construct an RNA-positive control based on armored (AR) RNA technology, using HIV-1 RNA as a model. Methods: The MS2 maturase, a coat protein gene (at positions 1765 to 1787) and HIV-1 pol gene were cloned into pET-32a plasmid. The prepared plasmid was transformed into Escherichia coli strain BL2 (DE3), and the expression of the construct was induced by 1 mM of isopropyl-L-thio-D-galactopyranoside (IPTG) at 37 °C for 16 h to obtain the fabricated AR RNA. The AR RNA was precipitated and purified using polyethylene glycol and Sephacryl S-200 chromatography. Results: The stability of AR RNA was evaluated by treatment with DNase I and RNase A and confirmed by transmission electron microscopy and gel agarose electrophoresis. Tenfold serial dilution of AR RNA from 101 to 105 was prepared; real-time PCR assays had a range of detection between 101 and 105, R2 value was 0.998, and the slope of the standard curve was -3.33. Conclusions: Prepared AR RNA, as a positive control, could be used as a basis for launching an in-house HIV-1 virus assay and other infectious agents. It can be readily available to laboratories and HIV research centers. The AR RNA is non-infectious and highly resistant to ribonuclease enzyme and can reduce the risk of infection in the clinical laboratory.
کلیدواژه‌های انگلیسی مقاله HIV-1, Real-time PCR, Virus-like particle
نویسندگان مقاله | Mohammad Gholami
Department of Medical Virology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran


| Mehrdad Ravanshad
Department of Medical Virology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran


| Kazem Baesi
Hepatitis andAIDS Department, Pasture Institute of Iran, Tehran, Iran


| Siamak M. Samiee
Food and Drug Laboratory Research Center, Ministry of Health and Medical Education, Tehran, Iran


| Negin Hosseini Rozbahani
Department of Immunology, Faculty of Medicine, Tehran University of Medical Sciences, Tehran, Iran


| Minoo Mohraz
Iranian Research Center for HIV AIDS (IRCHA), Iranian Institute for Reduction of High-Risk Behaviors, Tehran University of Medical Science, Tehran, Iran
نشانی اینترنتی http://ibj.pasteur.ac.ir/browse.php?a_code=A-10-1-730&slc_lang=en&sid=1
فایل مقاله اشکال در دسترسی به فایل - ./files/site1/rds_journals/125/article-125-931973.pdf
کد مقاله (doi)
زبان مقاله منتشر شده en
موضوعات مقاله منتشر شده Molecular Microbiology
نوع مقاله منتشر شده مقاله کامل
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