Exam 1 Terminology Flashcards
Virology
The study of viruses and viral diseases
Virologist
Someone who studies viruses
Virion
A complete virus particles that consists of an RNA or DNA core with a protein coat sometimes with external envelopes and that is the extracellular infective form of a virus.
Virus
Is a broad general terminology that is used to describe any aspect of the infectious agent and includes: the infectious (virion) or inactivated virus particles, or viral nucleic acid and protein in the infected cell.
Viroid
An infectious particle smaller than any of the known viruses, an agent of certain plant diseases. The particle consists only of an extremely small circular RNA (ribonucleic acid) molecule, lacking the protein coat of a virus.
Smallest viruses
Porcine circovirus type 1 (17 nm diameter)
Parvoviruses (18 nm diameter)
Largest viruses
Pandoravirus (400 nm diameter)
Poxvirus (200 nm) diameter and 300 nm in length)
Pleomorphism
The ability of some virus to alter their shape or size
Electron Microscopy (EM)
Requires negative staining with electron dense material. Resolution range is usually 50-75 angstroms.
Cryo-electrom Microscopy (Cryo-EM)
Allows the observation of biological specimens in their native environment (not stained or fixed in any way) at cryogenic temperatures in EM.
X-ray Crystallographic Method
The virus is crystallized. Then you bombarded the virus crystal with x-rays. The x-rays will strike and be reflected. Can measure angle of reflection and intensity of reflection. Fit into the computer and with software can reconstruct the virus structure.
Capsid
A protein (coat) shell of a virus that encases/envelopes the viral nucleic acid or genome
Capsomeres
Make up a capsid by non-covalent bonds. It is the basic subunit protein in the capsid of a virus.
Nucleocapsid
Capsid + virus nucleic acid
Triangulation number (T-number)
Described the relation between the number of pentagons and hexagons of the icosahedron. The larger the T-number the more hexagons are present relative to the pentagons.
T-number formula
T = (h^2 + h * k + k^2)
Glycoprotein
Spike embedded on the envelope. Anchored in the lipid bilayer by means of hydrophobic bonds. Transmembrane proteins.
Matrix protein
Between the lipid envelope and the capsid. These proteins link the internal nucleocapsid to the lipid membrane envelope.
External glycoprotein
Anchored in the envelope by a single transmembrane domain, and a short internal tail. Usually major antigens.
Hemagglutinin (HA)
Binding, fusion, antigenic, hemagglutination, clumping RBC
Neuraminidase (NP)
Release progeny virus from host cell, antigenic
Channel proteins
Which are most hydrophobic proteins that form a protein lined channel through the envelope. Alters permeability of the membrane.
Fusion proteins
Fuse to host cytoplasmic membrane. Facilitates virus into the host cell. Host cytoplasmic membrane and virus envelope is fused. Needed to penetrate into the host cell.
Positive sense RNA genome
It is similar to mRNA and thus can be immediately translated by the host cell and can directly synthesize viral proteins.
Negative sense RNA genome
Is complementary to mRNA and must be converted to positive sense RNA by an RNS polymerase before translation.
Antigenic Drift
Caused by point mutations. Mutation is a change of nucleotide of the genome. There is a change of antigenicity of the virus as a result of point mutation.
Antigenic Shift - Recombination
Intramolecular recombination involves the exchange of nucleotide sequences between different nucleotides. Usually by closely related, viruses during replication.
Antigenic Shift - Reassortment
Is the most important mechanism for high genetic diversity in viruses with segmented genome.
Lysins
Hydrolytic enzymes produced by bacteriophages to cleave the host’s cell wall
Retroviral integrase (IN)
Enzyme produced by a retrovirus (such as HIV) that enables its genetic material to be integrated into the DNA of the infected cells
Reverse Transcriptase (RT)
Enzyme used to generate complementary DNA (cDNA) from a RNA template
Nucleic acid polymerase
viral genome replication
Neuraminidases
Enzymes that cleave glycosidic bonds. Allows release of viruses from host cell.
Incomplete virions
Virion without nucleic acid (empty capsid)
Defective virions
a virus that cannot replicate because it lacks a full complement/copy of viral genes.
Defective Interfering Particle (DIP)
When the defective viruses cannot replicate, but can interfere other congeneric mature virion entering the cells, we call them defective interfering particles (DIP)
Pseudovirion
Contains non-viral genome within the viral capsid, such as host nucleic acid instead of viral nucleic acid. Look like ordinary viral particles under EM, but do not replicate.
Pseudotypes
When related viruses infect the same cell, the genome of one virus may be enclosed in the heterologous capsid of the second virus.
The Baltimore Classification System
Is a classification system based on viral genome. Viruses are classified into one of the seven groups depending on combination of their nucleic acid (DNA or RNA), strandness, sense, and method replication.
The International Committee Taxonomy of Viruses (ICTV) Classification System
The ICTV is the only body charged by the international union of microbiological societies with the task of developing, refining, and maintaining a universal virus taxonomy. Considers: nature of virus genome and virus genetic diversity, virus replication strategies, and virus morphology.
Monolayer cultures
When the bottom of the culture vessel is covered with a continuous layer of cells, usually one cell in thickness
Primary Cell Culture
Growth of cells dissociated directly from the parental tissue. Cells have the same chromosomes and same number of chromosomes as the original tissue.
Secondary (Transfer) Cell Culture
When a primary culture is sub-cultured. This is periodically required to provide fresh nutrients and growing space for continuously growing cell lines.
Finite/Diploid Cell Lines
Homogenous population of a single cell type - fewer cell types. Derived mainly from embryos; or from secondary cell culture. Cells retain original morphology and diploid chromosome number. Growth rate is slow.
Continuous Cell Lines
Divide indefinitely. Derived directly from cancer cells; or induce transformation of a primary of diploid cell strain to divide indefinitely. Abnormal morphology and chromosome number. FDA prohibits their use in vaccine production.
Phenol Red pH Indicator
Red is normal color, at a pH of 7-7.5. If there is an accumulation of toxic substances in medium then it will become acidic and it will become yellow or orange in color.
Carbon Dioxide (CO2) Level
Changes in atmospheric CO2 can alter the pH of the medium. Therefore, it is necessary to use exogenous CO2 when using media buffered with a CO2 bicarbonate based buffer.
How will you dissociate/deattach adherent cells?
Use enzyme trypsin which is a proteolytic enzyme and cause lysis of adhesion factors. Or use integrins which rely on calcium
Cytopathogenic Effect (CPE)
Refers to damage or morphological changes to host cells during virus invasion
Shell vial
Small borosilicate glass vial with a coverslip. Grow cells in coverslip or shell vial. Inoculate the virus into the vial. Virus grows in monolayers. Take cover slip out and drip it with fluorescent antibody and look under the microscope.
Cultivation of viruses in eggs
Egg candling and egg inoculation
When can you not use eggs for experiments?
If there is a crack in the eggshell, infertile egg, and blood ring indicated early embryonic death
Routes of egg inoculation
- Chorioallantoic membrane inoculation
- Amniotic inoculation
- Allantoic inoculation
- Yolk Sac inoculation
Yolk Sac Inoculation
Drill hole (use 22 gauge), place the inoculum below the embryo and within the yolk material. Seal the hole in the eggshell with scotch tape or melted wax.
Allantoic Cavity Inoculation
26 gauge, drill hole in eggshell above or over the air sac. Inoculate specimen into the allantoic cavity.
Amniotic Cavity Inoculation
26 gauge, drill a hole in the egg shell over the air sac, inoculate specimen into the amniotic cavity.
Chorioallantoic Membrane Inoculation (CAM)
Drill two holes in the eggshell, one on the side and the other above the air sac. Move air sac to the side of the egg by gentle suction, using a rubber bulb. Inoculate specimen on the dropped CAM.
Pocks on CAM
Most common lesions found in eggs. Assay is known as pocks assay. Pocks formed by vaccine virus on CAM.
Ultracentrifuge
Provide sufficient gravitational force to efficiently sediment even the smallest viruses
Rate-zonal centrifugation
Narrow zone on the top of a density gradient. Under centrifugal force, particles move at different rates depending on their mass.
Buoyant density
If the object has exactly the same density as the fluid, then its buoyancy equals its weight. It will remain submerged in the fluid, but it will neither sink nor float.
Isopycnic Point
The point where the buoyant density of a particle equals that of the surrounding density gradient medium.
Isopycnic Centrifugation
Particles are separated solely on the basis of their buoyant density. Particles will never sediment to the bottom of the tube. Virus particles appear as visible bands.
Virus Purification with Membrane Chromatography
Chromatography membranes are not utilized in purification of viruses and virus like particles and for impurity removal. The macroporous structure of the membrane allows large viruses to enter it and to bind to the inner pore surface easily.
Binding of the viruses on ion exchange membranes depends on charge distribution on the virus. Adsorption and subsequent elation of viruses can be achieved by anion and cation exchange membranes.
Virus Quantification
Counts the number of viruses in a specific volume to determine the virus concentration
Virus Titer
The lowest concentration of virus that still infects cells
Biological - Viral Quantification Tests
Depend on a virus particle initiating a successful replication cycle. Plaque assays, pock assays, various endpoint titration methods.
Physical - Viral Quantification Tests
Do not depend on any biological activity of the virus particle. More related on the counting. EM, Hemagglutination, ELISA, PCR, flow cytometry.
Transmission Electron Microscopy (TEM)
Direct particle counts; the most direct method to determine the concentration of virus particles in a sample.
Virus Counter 2100
The technology behind the virus counter 2100 is based on specialized version of flow cytometry developed specifically for use with nanometer scale particles.
When using a virus counter …
Each sample is stained with two different fluorescent dyes, one specific for nucleic acid, and the other specific for protein, and analyzed as they flow through a laser beam
Hemagglutination Assay
Antigen concentration assessment. RBC in suspension becomes linked (agglutinated) by ND virus receptors. RBC settle to form a mat.
High Performance Liquid Chromatographhy (HPLC)
Antigen concentration assessment. The concentrations of specific viral antigens may be quantified through UV analysis of fractions generated during HPLC.
Single Radial Immunodiffusion (SRID)
Antigen concentration assessment. Radial diffusion of purified viral antigens (standards) and viral particles through agarose gel seeded with polyclonal antisera against a viral antigen.
Monolayer Plaque Assay
A circular zone of white necrotic cells surrounded by viable cells in a monolayer. It is a functional measurement, and has no relation to actual number of viruses.
Plaque Forming Units/ml (PFU)
Measures the number of virus particles capable of forming plaques per unit volume.
Transformation Assay
Quantitative determination of titers of oncogenic viruses. Oncogenic viruses transform cells in culture. Transformed cells lose contact inhibition and become heaped upon one another.
Quantal Assay
Measures the presence of absence of infection. Use for certain viruses that do not form plaques or for determining the virulence of a virus in animals or eggs.
Lethal dose 50
Amount of virus that kills 50% of test subjects
TCID50
A Tissue Culture Infectious Dose which will infect 50% of the cell monolayers challenged with the defined inoculum
Multiplicity of Infection (MOI)
Is the average number of virus particles infecting each cell
TCID50 Formula
(% Infection above 50% - 50%)/ (% Infection above 50% - % Infection below 50%)
Permissive Cell
A cell in which a virus is able to replicate
Non-permissive cell
Cells in which a factor or factors necessary to viral reproduction is not present or one detrimental to viral reproduction is present
One-step Virus Growth Curve
- Viral adsorption and entry
- Viral uncoating and replication
- Viral maturation
- Viral release
Burst Size
Number of infectious virions released per average cell
Adsorption
During this period, virus attached to and enters cells, and the titer of free virus in the medium may actually decline
Eclipse Period
Tie interval between uncoating (“disapperance” of viruses) and appearance, intracellularly, of first infectious progeny virion.
Latent Period
The time before new infectious virus appears in the medium. I.e., from uncoating to just prior to the release of the first extracellular virions.
Steps of Virus Replication
- Attachment (adsoprtion)
- Penetration (Injection)
- Uncoating
- Synthesis of viral components (nucleic acid and protein)
- Assembly and maturation
- Release in large numbers (lysis)
Co-receptor
In some cases, binding to a cellular receptor is not sufficient for infection. An additional cell surface molecule, or co-receptor, is required for entry.
Endocytosis
A process in which a substance gains entry into a cell without passing through the cell membrane. The process involves invagination and pinching off of small regions of the cell membrane, resulting in the non-specific internalization of molecules.
Surface Fusion
Type of virus penetration. Only enveloped viruses. Membrane fusion or fusion proteins facilitates membrane fusion.
pH dependent surface fusion and uncoating
Low pH in endosome promotes fusion of envelope with endosomal membrane; lysis of nucleocapsid by lysosomal proteases, and release of viral genome
pH independent surface fusion and uncoating
The F protein catalyzes membrane fusion at the cell surface at neutral pH. The viral nucleocapsid is then released into the cytoplasm.
Pore-mediated penetration
Penetration of viral genome into host cell (non-enveloped aka naked viruses). Inject their genome into the host cytoplasm through creation of a pore in the host membrane.
Antibody-mediated penetration
Antibody (Ig - Fc) receptor enhances the entry of viruses into host cell
Capping
Addition of 7-methylguanosine of the 5’ end of RNA. Provides stability, binding of mRNA to ribosomes, and mark mRNA as “self”.
3’-Polyadenylation
Viral mRNAs can be polyadenylated by host or viral enzymes. A stretch of adenylated residues are added to the 3’ end.
Splicing
RNA splicing is a process that removes introns and joins exons in a primary transcript
Exon
the portion of a gene that codes of amino acids
Intron
a portion of a gene that does not code for amino acids
Constituitive splicing
Every intron is spliced out; every exon is spliced in
Alternative splicing
All introns spliced out; only selected exons spliced in. Results in mRNAs having different coding information derived from a single gene
Monocistronic
A type of viral mRNA that encodes one polypeptide. Singe RNA that will encode one protein.
Polycistronic
A type of viral mRNA that encodes several polypeptides.
Exocytosis
By budding through the membrane of golgi apparatus or ER. Vesicles containing the virus then migrate to the plasma membrane and are released by exocytosis
Extracellular Spread
Released viruses in extracellular environment. Travel to a new cell. Infect new host cell. Same replication cycle repeated etc.
Intercellular Spread
Spread from cell-to-cell without contact with extracellular environment. Results in rapid virus dissemination, evasion of immune system and persistent infections
Disseminated infection
Infection spreads beyond the primary site of infection
Systemic infection
If a number of organs or tissues are infected
Viremia
The presence of a virus in the blood. Virus may be free in the blood or in a cell.
Passive viremia
Direct inoculation of virus in the blood. Bite of arthropods or contaminated syringe
Primary viremia
Initial entry of virus into the blood after infection