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Methods for the Study of HIV-1 Assembly

Virus assembly constitutes a key phase of the HIV-1 replication cycle. The assembly process is initiated by the synthesis of the Gag precursor protein, Pr55Gag, in the cytosol of the infected cell. After its synthesis, Pr55Gag is rapidly transported in most cell types to the plasma membrane (PM) wh ...

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Assembly of Immature HIV-1 Capsids Using a Cell-Free System

For many years it has been known that viral capsid proteins are capable of self-assembly, but increasing evidence over the past decade indicates that in cells HIV-1 capsid assembly occurs via a complex but transient series of steps requiring multiple viral–host interactions. To better unde ...

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Wild Bird Surveillance for the Avian Influenza Virus

Avian influenza (AI) viruses have been isolated from a wide diversity of free-living avian species representing several orders. Isolations are most frequently reported from aquatic birds in the Orders Anseriformes and Charadriiformes, which are believed to be the reservoirs for all AI ...

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Immunohistochemical Staining for the Detection of the Avian Influenza Virus in Tissues

Immunohistochemical methods are commonly used for studying the pathogenesis of the avian influenza (AI) virus by allowing the identification of sites of replication of the virus in infected tissues and the correlation with the histopathological changes observed. In this chapter, t ...

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Reverse Genetics of the Avian Influenza Virus

Reverse genetics is the creation of a virus from a full-length cDNA copy of the viral genome, referred to as an infectious clone, and is the most powerful genetic tool in modern virology. The generation of influenza A viruses by reverse genetics has lagged mainly due to the inherent technical difficul ...

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Evaluating the Cell-Mediated Immune Response of Avian Species to Avian Influenza Viruses

The measurement of avian cellular immunity is critical to understanding the role and regulation of avian lymphocytes following avian influenza (AI) virus infection. Although the ability to measure avian T cell responses has steadily increased over the last few years, few studies have ex ...

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Measurement of Avian Cytokines with Real-Time RT-PCR Following Infection with the Avian Influenza Virus

Functional and molecular techniques have both been employed to measure the production of cytokines following influenza infection. Historically, the use of functional or antibody-based techniques was employed in mammalian immunology. In avian immunology, only a few commercial ...

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A Brief Introduction to the Avian Influenza Virus

The avian influenza (AI) virus is type A influenza isolated from and adapted to an avian host. Type A influenza belongs to the orthomyxovirdae virus family, is enveloped, and is pleiomorphic with a size ranging from 80–120 nm (reviewed in ). Type A influenza strains are classified by the serological su ...

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Avian Influenza Virus RNA Extraction from Tissue and Swab Material

The efficient extraction and purification of viral RNA are critical for downstream molecular applications, whether it is the sensitive and specific detection of virus in clinical samples, virus gene cloning and expression, or quantification of the avian influenza (AI) virus by molecu ...

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Avian Influenza Virus Sample Types, Collection, and Handling

Successful detection of the avian influenza (AI) virus, viral antigen, nucleic acid, or antibody is dependent upon the collection of the appropriate sample type, the quality of the sample, and the proper storage and handling of the sample. The diagnostic tests to be performed should be consider ...

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Type A Influenza Virus Detection and Quantitation by Real-Time RT-PCR

Real-time RT-PCR (rRT-PCR) is a relatively new technology that has been used for avian influenza (AI) virus detection since the early 2000s for routine surveillance, during outbreaks, and for research. Some of the advantages of rRT-PCR are high sensitivity, high specificity, rapid time-to- ...

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Detection and Identification of the H5 Hemagglutinin Subtype by Real-Time RT-PCR

Serological methods, gene sequencing, and RT-PCR-based methods have all been used for the identification of influenza virus hemagglutinin (HA) subtypes. Compared to serological methods and gene sequencing, RT-PCR is fast, sensitive, and relatively inexpensive. However, since RT ...

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Avian Influenza Virus Isolation and Propagation in Chicken Eggs

The avian influenza (AI) virus is usually isolated and propagated by inoculating either swab or tissue samples from infected birds into the chorioallantoic sac of embryonating chicken eggs. This is the accepted method, but occasionally an isolation may only be successful when inoculat ...

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Hemagglutination Assay for the Avian Influenza Virus

The hemagglutination (HA) assay is a tool used to screen cell culture or amnioallantoic fluid harvested from embryonating chicken eggs for hemagglutinating agents, such as type A influenza. The HA assay is not an identification assay, as other agents also have hemagglutinating properti ...

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Hemagglutination-Inhibition Test for Avian Influenza Virus Subtype Identification and the Detection and Quantitation of Serum Antibodies to the Avian

The hemagglutination-inhibition (HI) assay is a classical laboratory procedure for the classification or subtyping of hemagglutinating viruses. For the avian influenza (AI) virus, the HI assay is used to identify the hemagglutinin (H) subtype of an unknown AI virus isolate or the HA subty ...

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Neuraminidase-Inhibition Assay for the Identification of Influenza A Virus Neuraminidase Subtype or Neuraminidase Antibody Specificity

The neuraminidase-inhibition (NI) assay is a laboratory procedure for the identification of the neuraminidase (NA) glycoprotein subtype in influenza viruses or the NA subtype specificity of antibodies to influenza virus. A serological procedure for subtyping the NA glycoprote ...

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Detection and Quantification of Serum or Plasma HCV RNA: Mini Review of Commercially Available Assays

The treatment schedule (combination of compounds, doses, and duration) and the virological follow-up for management of antiviral treatment in patients chronically infected by HCV is now well standardized, but to ensure good monitoring of the treated patients, physicians need rapid, ...

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Simultaneous Detection of Anti-HCV Antibody and HCV Core Antigen

HCV infection is usually diagnosed by means of an enzyme immune assay for the detection of antibody against HCV. The window period between infection and seroconversion remains a dramatic problem in the transfusional and diagnostic setting. In this chapter, we report (i) procedures for assa ...

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Immunohistochemical Detection of HCV Proteins in Liver Tissue

Detection and localization of HCV in liver tissue are vital for diagnostic purposes and clinical management of HCV-infected patients, as well as for the elucidation of viropathological mechanisms. The fragility of HCV RNA and the low levels of viral expression in infected tissues are a cons ...

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Nomenclature and Numbering of the Hepatitis C Virus

International standardization and coordination of the nomenclature of variants of hepatitis C virus (HCV) is increasingly needed as more is discovered about the scale of HCV-related liver disease and important biological and antigenic differences that exist between variants. C ...

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