ABSTRACT
In order to establish a rapid and accurate method for the detection of Ebola virus (EBOV), the primers used in SYBR Green I real-time RT-PCR were designed based on the EBOV NP gene sequences published in GenBank. The SYBR Green I real-time RT-PCR was established and optimized for the detection of EBOV. The EBOV RNA that was transcribed in vitro was used as a template. The sensitivity of this method was found to reach 1.0 x 10(2) copies/microL and the detection range was 10(2) - 10(10). No cross reaction with RNA samples from Marburg virus, Dengue virus, Xinjiang hemorrhagic fever virus, Japanese encephalitis virus, Influenza virus (H1N1 and H3N2) and Porcine reproductive and respiratory syndrome virus E genomic RNA was found. The method would be useful for the detection and monitoring of EBOV in China.
Subject(s)
Humans , DNA Primers , Chemistry , Genetics , Ebolavirus , Genetics , Hemorrhagic Fever, Ebola , Virology , Organic Chemicals , Chemistry , Reverse Transcriptase Polymerase Chain Reaction , MethodsABSTRACT
ISG15 is a 15kD ubiquitin-like protein (UBL) induced by interferon (IFN). ISG15 can be covalently attached to proteins, which is called ISGylation process. ISGylation system contains ISG15, UBE1L, UBCH8 and HERC5 proteins, which are all essential for ISGylation. ISG15 and ISGylation system have been found to have anti-viral effects. A better understanding of how ISG15 mediates the anti-viral activity will provide insights for new anti-viral drugs development and new therapeutic strategies. The mechanisms underlying the ISG15 mediated anti-viral response have been explored extensively in recent years. This minireview summarized the research advances of how ISG15 mediated the anti-viral effects against different kinds of viruses.
Subject(s)
Animals , Humans , Cytokines , Physiology , HIV Infections , Allergy and Immunology , Influenza, Human , Allergy and Immunology , Ubiquitins , Physiology , Virus Diseases , Allergy and ImmunologyABSTRACT
Classical swine fever (CSF) is a contagious swine disease charactered by hemorrhagic fever and leukopenia,usually leading to substantial economic losses. To obtain a insight of leucopenia caused by CSFV infection, DNA microarray analyses of peripheral blood leucocytes (PBL) of the infected pigs was performed. Three health pigs were inoculated with a lethal dose of CSFV Shimen strain and their PBLs were isolated when the onset of typical clinical signs and then subjected to total RNA extraction followed by microarray analysis with Affymetrix Porcine Genome Array GeneChips. The results showed that the significant differences were observed in cellular apoptotic genes expression at 7 days post-infection (p. i.). The changes of the genes expression were confirmed by real time RT-PCR of some selected apoptosis-related genes. This study provided a valuable information for further investigating the molecular mechanism of apoptosis caused by CSFV infection.