Which Of The Following Is Not Used To Culture Viruses

chapter 13 Flashcards – Easy Notecards

1How do all viruses and bacteria vary from one another? Viruses are not made up of cells like other organisms. Identify which of the following assertions gives the most compelling evidence in favor of the notion that viruses are nonliving substances. They are unable to replicate themselves unless they are in the presence of a host. Identify which of the following assertions regarding viral spikes is FALSE. They can only be discovered on viruses that are not enclosed. In the classification of viruses, which of the following is NOT utilized as a criteria is not used?

In which of the following processes do Bacteriophages and animal viruses not differ significantly from one another?

10The structures depicted in Figure 13.1 are made up of the following components: 11A clean region that protects against a confluent lawn of germs is referred to as a 12An important distinction between continuous cell lines and primary cell lines is that continuous cell lines may be maintained for an endless number of generations, but primary cell lines cannot.

1414) A persistent infection is one in which the illness process progresses slowly over an extended period of time.

  • The prophage protects the host cell from infection by other phages by making it immune to them.
  • What would be the initial stage in the biosynthesis of a virus with a (minus) strand of RNA if it were one of the following?
  • 19An envelope is produced during which of the following steps?
  • It results in the lysis of host cells.
  • 22The capacity of a virus to infect a cell in an animal is mostly dependent on the availability of receptor sites on the cell’s membrane.
  • This virus has a single RNA strand that is positive in the sense direction, as well as an envelope.
  • 24 The process by which an enveloped virus escapes from a host cell is referred to as 25 Cancer that develops after the injection of cell-free filtrates provides the most solid proof that viruses are responsible for cancer.

EXCEPT27 It is only during bacteriophage reproduction that naked nucleic acid is introduced into the host cell, which distinguishes it from viral replication in other organisms.

Which of the following correctly arranges these components in the proper sequence for DNA virus replication?

The process of maturation 2.

Transcription is another option.

Interpretation30 In the world of viruses, a viral species is a group of viruses that share genetic information and an ecological habitat.

Hepadnaviridae and Retroviridae are two families of viruses.

Viruses make use of catabolic enzymes that they produce on their own.

Which of the following would be the most accurate description of what is taking place within the patient?

35Some viruses, such as the human herpesvirus 1, can infect a cell without creating any symptoms in the recipient.

As a 60-year-old woman suffering from shingles (human herpesvirus 3), which line on the graph in Figure 13.2 would represent the number of viruses present in this individual?

In which period of time on the graph in Figure 13.2 would the patient exhibit the signs and symptoms of the disease?

Which of the following is the third step?

Which of the following is the fourth step?

42 Which of the following is most likely to be a product of an early gene in the human population?

Which of the following enzymes is found in the majority of RNA viruses? 44The following stages take place during the biosynthesis of an RNA virus with a positive strand. What is the third phase in this process? 45What factors have a role in the antigenic change of influenza viruses?

Culturing Viruses

Bacteriophage cultures necessitate the use of host cells in which the virus or phage may reproduce.

Learning Objectives

Explain why and how to batch culture bacteriophages, as well as the benefits of doing so.

Key Takeaways

  • A bacteriophage is a kind of virus that infects bacteria. It accomplishes this by injecting genetic material – either DNA or RNA – into the cell, which it transports in an outer protein capsid. Bacteriophages attach themselves to certain receptors on the surface of bacteria, such as lipopolysaccharides, teichoic acids, proteins, or even flagella, in order to gain entry into the host cell. The movement of phage virions in solution is dependent on chance contacts with the appropriate receptors (blood, lymphatic circulation, irrigation, soil water, and so on)
  • When in solution, they must rely on random interactions with the appropriate receptors.

Key Terms

  • Bacteriophage: A virus that infects bacteria alone
  • Also known as bacteriophage.

Strategies of Replication

In order for virus or phage cultures to proliferate, they must be housed in host cells. Bacteriophage cultures are created by infecting bacterial cells with the bacteriophage. The phage may then be separated from the plaques that form in a lawn of bacteria on a plate by using a phage isolation kit. Bacteriophages infecting a bacterium are classified as follows: In order for virus or phage cultures to proliferate, they must be housed in host cells. Bacteriophage cultures are created by infecting bacterial cells with the bacteriophage.

  • A bacteriophage is any one of a variety of viruses that infect bacteria, and there are many different types.
  • The genetic material can be ssRNA, dsRNA, ssDNA, or dsDNA (the prefix’ss-‘ or ‘ds-‘ implies single-stranded or double-stranded DNA, respectively), and it can be arranged in either circular or linear fashion.
  • Bacteriophages attach themselves to certain receptors on the surface of bacteria, such as lipopolysaccharides, teichoic acids, proteins, or even flagella, in order to gain entry into the host cell.
  • The capacity of the phage to adhere to and infiltrate the host is also influenced by the growing circumstances of the host.
  • Phages can be discharged in a variety of ways, including cell lysis, extrusion, and, in a few instances, budding.
  • The filamentous phages, which are a completely distinct form of phage, cause the host cell to release additional virus particles on a continuous basis.
  • Several Mycoplasma phages are connected with the process of budding.

Tissue Culture of Animal Viruses

Instead of growing viruses in ordinary microbiological broth or on agar plates, viruses must be cultivated within host cells that are compatible with their growth.

Learning Objectives

Learn about the benefits of cultivating animal viruses in host cells as well as the reasons for doing so.

Key Takeaways

  • In order to cultivate clinical samples that are suspected of carrying a virus, tissue culture is a valuable technique. This approach is useful in the laboratory for the detection, identification, and characterisation of viruses
  • It is also useful in the field. Growing animal cells in flasks using different broth medium and subsequently infecting these cells with virus is the process of tissue culture of animal viruses. Calcium phosphate, electroporation, or combining a cationic lipid with the material to generate liposomes, which fuse with the cell membrane and deposit their payload within, are all methods for carrying out transfection. When viruses infect cells, they induce non-lytic damage, which is known as cytotoxicity. These present themselves in a variety of ways and have a variety of negative consequences.

Key Terms

  • In cell culture, a sophisticated process through which cells are cultivated under controlled circumstances, usually outside of their natural habitat, is called cell culture. The term “cytopathic effect” refers to the degenerative changes that occur in cells, particularly in tissue culture, and is sometimes connected with the replication of some viruses.

Generally speaking, cell culture is a complicated process in which cells are cultivated under controlled circumstances, usually outside of their native habitat. In practice, the word “cell culture” now refers to the cultivation of cells originating from multicellular eukaryotes, particularly animal cells, rather than the cultivation of single cells. There are, however, cultures of plants, fungi, and microorganisms, including viruses, bacteria, and protists, that have been created. It is important to note that the historical history and current methods of cell culture are intimately tied to those of tissue and organ culture.

  1. Viruses are intracellular parasites that require the presence of live cells in order to reproduce and spread.
  2. Despite the fact that embroyonated eggs and laboratory animals are extremely beneficial for the isolation of specific viruses, cell cultures are the only approach used in the majority of laboratories for viral isolation.
  3. In order to produce cell cultures, tissue pieces must first be dissociated, which is commonly accomplished with the use of trypsin or collagenase enzymes.
  4. Examples include Eagle’s and an animal serum.
  5. The attachment of normal cells to a stable support is required for their growth to be successful.
  6. The sensitivity of various cell cultures to different viruses varies significantly.
  7. As soon as feasible after collection, samples for cell culture should be delivered to the laboratory for further processing.
  8. Fluids and tissues from the body should be collected and stored in a sterile container.
  9. Change the maintenance media after one hour or the next morning, whichever comes first.
  10. Rotation is the most effective method of isolating respiratory viruses, and it also results in the manifestation of cytopathic effects (CPE) for many viruses occurring early in the isolation process.

If stationary tubes are utilized, it is vital that the culture tubes are positioned in such a way that the cell monolayer is bathed in nutritious media. If this is not done, the cells will die.

Inoculation of Live Animals

Cultivation of cells is a complicated process in which cells are cultivated under controlled circumstances, usually outside of their native habitat. It is now well understood that the word “cell culture” is referring to the process of growing and cultivating cells that have been obtained from multicellular eukaryotes, particularly animal cells. There are, however, cultures of plants, fungi, and microorganisms, including viruses, bacteria, and protists, that may be found in the laboratory. It is important to note that the historical history and current methods of cell culture are intimately linked to those of tissue and organ culture.

  1. Viruses are intracellular parasites that require the presence of live cells in order to reproduce successfully.
  2. Cell cultures are the only technique used in most laboratories for viral isolation, despite the fact that embroyonated eggs and laboratory animals are quite beneficial for specific viruses.
  3. Typically, trypsin or collagenase are used to separate tissue pieces prior to culturing them in cell culture.
  4. Example: an animal serum such as Eagle’s and a fish serum It is possible that the cells may connect and spread on the bottom of the container after a variable lag time and then begin to divide, resulting in the development of a primary culture.
  5. Effect on the cytotoxic immune system (cytotoxicity): Herpes simplex virus has a cytotoxic effect on cells.
  6. If a suspected virus is detected in a cell culture, it is critical that the most sensitive cell culture be employed.
  7. A vial containing viral transport medium should be placed in which the swabs will be placed.
  8. When the material is received, it is injected into a variety of various types of cell culture, which are selected based on its nature and clinical presentation.
  9. Incubate the infected tubes at 35-37 degrees Celsius in a revolving drum for at least 24 hours.
  10. It also results in the manifestation of cytopathic effects (CPE) for many viruses occurring earlier than would otherwise be the case.

For cell monolayers to be bathed in nutritional medium, it is vital that the culture tubes are positioned so that the cell monolayer is immersed in the nutrient media.

Learning Objectives

Describe the process of innoculation of live animals.

Key Takeaways

  • It was on human volunteers that live inoculation of yellow fever virus was first utilized for research purposes
  • The most commonly employed animals in viral cultivation include primates such as primates and monkeys, rabbits, guinea pigs, rats, hamsters, and mice
  • Live animal inoculation necessitates the use of skilled workers who are familiar with the different injection procedures.
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Key Terms

  • The term latent refers to something that is existing or exists but is hidden or inactive. The process of tumor creation and development (oncogenesis). The introduction of an antigenic material or vaccine into the body in order to create immunity to a certain illness is known as vaccination.

In this micrograph of the yellow fever virus, you can see the virus’s shape. Viruses are obligatory intracellular parasites that cannot survive in the absence of an active host. The virus requires living cells in order to reproduce, which can be given through injection in live animals, among other means of virus cultivation (cell culture or inoculation of embryonated eggs). When it came to virus culture and studying viral illness, the only way that was known was to infect healthy volunteers with viruses.

  1. Human volunteers are only recruited when there is no alternative technique available and the virus is reasonably safe, due to the high level of danger involved.
  2. It was also the first disease for which a vaccine was developed.
  3. It has only been a few decades since animal inoculation has been used to isolate viruses from their hosts.
  4. This is because the kind of virus to be isolated has a significant influence on the choice of animals and method of injection (intracerebral, intraperitoneal, subcutaneous, intradermal, or intraocular).
  5. Furthermore, animal inoculation may be utilized to study pathogenesis, immune response, epidemiology and oncogenesis in addition to viral isolation and isolation studies.
  6. It is possible that many passages into animals will be necessary to produce observable signs of viral growth in some cases.

Viral Identification

There is a significant difference in the genetic material contained within viral particles between various kinds of viruses.

Learning Objectives

Examine the differences between the replication of DNA, RNA, and reverse transcribing viruses.

Key Takeaways

  • The nucleus of the cell is where the majority of DNA viruses replicate their genomes. RNA viruses may be classified into four kinds based on how they replicate
  • The first two are the most common. Retroviruses, Metaviridae, and Pseudoviridae are reverse transcribing viruses that include ssRNA (Retroviridae, Metaviridae, Pseudoviridae) or dsDNA (Caulimoviridae, and Hepadnaviridae) in their particles
  • Reverse transcribing viruses are also known as reverse transcription viruses.

Key Terms

  • Genome: An organism’s whole genetic information (either DNA or, in the case of certain viruses, RNA), which is normally represented in basepairs.

Replication of Viruses

Variations in the genetic material contained within viral particles, as well as the manner by which the genetic material is replicated, exist amongst various types of viruses.

TYPES

DNA viruses: The genome replication of the vast majority of DNA viruses takes place in the nucleus of the host cell. In certain cases (for example, herpesviruses), these viruses can enter the cell directly by fusing with the cell membrane (as in the case of herpesviruses), but most of the time they do so by receptor-mediated endocytosis. Most DNA viruses are completely reliant on the DNA and RNA synthesizing and RNA processing equipment found in the host cell; but, viruses with bigger genomes may be able to encode a significant portion of this machinery for themselves.

  1. RNA viruses are classified as follows: The majority of the time, replication takes place in the cytoplasm.
  2. In addition, the genetic material must be single-stranded or double-stranded, which is an important distinction.
  3. Reconstruction of a rotavirus particle using computer-assisted techniques: Baltimore is a good example.
  4. IV: () a positive number ssRNA viruses are a kind of virus that is characterized by the presence of a single strand of RNA.
  5. ssRNA-RT viruses are the sixth kind of virus.
  6. Viruses that reverse transcription: These viruses have ssRNA (Retroviridae, Metaviridae, Pseudoviridae) or dsDNA (Caulimoviridae, and Hepadnaviridae) in their particles, but the other viruses do not have either.
  7. When it comes to nucleic acid conversion, both forms make use of a reverse transcriptase (also known as an RNA-dependent DNA polymerase enzyme).
  8. Pararetroviruses do not, despite the fact that integrated genome copies, which are often seen in plant pararetroviruses, can result in the production of infectious viruses.
  9. HIV, which is a retrovirus, is an example of the first type of virus.
  10. It was established by David Baltimore and is a viral classification system that divides viruses into families based on the kind of genome (DNA, RNA, single-stranded (ss), double-stranded (ds), and other factors) and the mode of reproduction used by the virus.

A major advantage of classifying viruses according to their genomes is that all of the viruses within a specific category will act in a similar manner, which provides some guidance on how to proceed with additional study. In conclusion, I would want to say that

  • I: dsDNA viruses (e.g. Adenoviruses, Herpesviruses, Poxviruses)
  • II: ssDNA viruses (+)sense DNA (e.g. Parvoviruses)
  • III: dsDNA viruses (e.g. Adenoviruses, Herpesviruses, Poxviruses)
  • IV: dsDNA viruses (e.g. dsRNA viruses are the third kind of virus (e.g. Reoviruses) Viruses with (+)ssRNA and (+)sense RNA (IV) (e.g. Picornaviruses, Togaviruses) Orthomyxoviruses, Rhabdoviruses, and other viruses with (ssRNA) sense RNA are examples of V viruses. VI: ssRNA-RT viruses (+)sense RNA with DNA intermediate in life-cycle (e.g. Retroviruses)
  • VII: dsDNA-RT viruses (e.g. Hepadnaviruses)
  • VIII: ssRNA-RT viruses (e.g. Retroviruses)
  • IX: ssRNA-RT viruses (e.g. Hepadnaviruses)
  • IX: ssRNA-RT viruses (e.g. Hepad

Ch. 6 Multiple Choice – Microbiology

In the first category are dsDNA viruses (e.g. Adenoviruses, Herpesviruses, Poxviruses); in the second category are positive (positive) sense DNA viruses (e.g. Parvoviruses); in the third category are dsDNA viruses (e.g. Adenoviruses); in the fourth category are dsDNA viruses (e.g. Adenovirus). viruses with double stranded RNA (e.g. Reoviruses) Viruses with (+)ssRNA and (+)sense RNA (e.g. Picornaviruses, Togaviruses) Orthomyxoviruses, Rhabdoviruses, and other viruses with (ssRNA) sense RNA (e.g.

Retroviruses); VII: dsDNA-RT viruses (e.g.

Retroviruses); VI: ssRNA-RT viruses (e.g.

Select all of the options that apply.

  1. Ribosomes, metabolic processes, nucleic acid, and glycoprotein are all terms that come to mind.

In addition, the envelope of a virus is generated from the host’s envelope. 4.When naming viruses, the family name finishes with the letter _, and the genus name ends with the letter _. 5.

  1. Virus
  2. Viridae
  3. Virus
  4. Virion

5.What is another term for a virus that is not enveloped? 6) Which of the following causes the apoptosis (death of the host cells)? 6) Which of the following causes the apoptosis (cell death) of host cells? In order for HIV to enter a cell, one of the following components must be introduced.

  1. A DNA polymerase that requires DNA as a cofactor
  2. An RNA polymerase
  3. A ribosome
  4. A reverse transcriptase

9.A virus with a positive strand of RNA:

  1. Prior to being translated, it must first be transformed into a messenger RNA (mRNA)
  2. It can be utilized directly to translate viral proteins
  3. It will be destroyed by host enzymes
  4. It is not recognized by host ribosomes

In what language is the transfer of genetic information from one bacteria to another bacterium known as phage transmission described? The following items cannot be used to cultivate viruses: 11, 12, 13, 14, 15, 16, 17, 18, 19, 20. 12.Which of the following tests may be performed to determine whether or not a certain virus is present in the body? 13.Which of the following does not have a cytopathic impact on the patient?

  1. Inclusion bodies
  2. Cell fusion
  3. Transformation
  4. Mononucleated cell
  5. Inclusion bodies

14.Which of the following pathogenic agents is devoid of nucleic acids? 15.Which of the following statements is correct about prions?

  1. They can be rendered inactive by boiling at 100 degrees Celsius. They have a capsid in them. This protein, PrP, is a renegade version of the protein. The use of an autoclave may be relied upon to inactivate them.

SOLVED:Which of the following is NOT utilized to culture viruses? culture media embryonated eggs laboratory animals animal cell cultures bacterial cultures Question 26 (1 point) Which of the following represents a virus family name? Herpes simplex virus Herpesviridae Picornavirus Enterovirus Hepatitis B virus

The ingredients of life are the fundamental building pieces that make up all living organisms, according to biology. Carbon, hydrogen, nitrogen, oxygen, phosphorus, and sulfur are the elements that make up the periodic table. The first four of these are the most significant since they are used to create the molecules that are required to form live cells. The other three are less important. These components serve as the fundamental building blocks of the primary macromolecules of life, such as carbohydrates, lipids, nucleic acids, and proteins, among other things.

  1. Carbon is found in all living things.
  2. Additionally, carbon is utilized in the construction of the energy-dense molecules adenosine triphosphate (ATP) and guanosine triphosphate (GTP) (GTP).
  3. Hydrogen is also required for the synthesis of ATP and GTP.
  4. It is also involved in the formation of ATP and GTP.

It is also involved in the formation of ATP and GTP. Carbohydrates, lipids, and nucleic acids are only a few of the fundamental building blocks of life that phosphorus is involved in the formation of.

Traditional and Modern Cell Culture in Virus Diagnosis

Osong Public Health Research Perspectives, Vol. 7, No. 2, April 2016, pp. 77–82. Several other papers in PMC have mentioned this article in their own work.

Abstract

It is necessary to generate cell cultures from tissue samples and then disaggregate them using mechanical, chemical, and enzymatic procedures in order to recover cells that are appropriate for viral isolation. Because of recent technological advancements, cell culture is now regarded to be the gold standard for viral isolation. This study examines the development of cell culture technologies over time and illustrates why cell culture is the favored approach for viral detection. It is also highlighted what are the pros and disadvantages of both conventional and current cell culture methods for diagnosing each type of virus, as well as the differences between the two.

However, it is necessary to incorporate certain more exact approaches, such as molecular methods, into cell culture in order to ensure the correct identification of viruses.

1. Introduction

During the early 1900s, embryonated eggs and laboratory animals were utilized to isolate viruses from their host organisms. A typical procedure involves the development of cell cultures from tissue samples, which are subsequently disaggregated using mechanical, chemical, and enzymatic procedures to obtain cells appropriate for viral isolation. The use of laboratory animals in investigations has dropped dramatically as a result of the development of the cell culture technology and its application.

The isolation of viral pathogens in cell cultures began in the 1960s; however, there were certain constraints at the time, including a lack of services for the diagnosis of viral infections and a lack of funding.

Many human viruses were successfully produced in vitro after the discovery of cell culture.

This approach is widely regarded as the gold standard for the isolation and identification of viruses.

2. Traditional cell culture for virus diagnosis

It was not until 1913 that a virus (vaccinia) was successfully grown in cell culture for the first time, and it wasn’t until the 1930s that viruses such as yellow fever and small pox were successfully cultivated in cell culture with the goal of producing vaccines. 3,4,5. Although the first virus (poliovirus) was discovered in 1950, it was not until then that it was isolated. It was possible to establish cell culture by adjusting the concentration of an antibiotic to avoid contamination with bacteria and adding certain chemicals to the media that served as the cell culture medium.

Cell culture may be conducted in any container, however, the common container is a screw-cap tube glass (16 mm × 125 mm;Figure 1) in which monolayer cells can grow on one side of the glass.

The main rhesus monkey kidney cells (RhMK), primary rabbit kidney cells (MRC-5), human foreskin fibroblasts (HEp-2), and A549 are the most significant cell lines frequently utilized for viral diagnostics.

2.1. Sample collection

The specimen processing methodology varies from laboratory to laboratory, however the following are the essential processes that must be followed: First, the medium holding the sample is vortexed, and the swap is removed from consideration. After that, the liquid medium is centrifuged. A cell culture procedure is carried out using the supernatant produced from the extraction process. This approach results in the retention of fungi, cells, bacteria, and blood in the bottom of the tube (in pellet form), but viruses stay scattered throughout the liquid.

  1. A 90-minute incubation at 35°C and 5 percent CO2 is then performed on the cell culture tube containing the virus for absorption, after which the inoculum is removed and replaced with new media.
  2. Depending on the infection, this procedure might take anything from a few hours to many weeks.
  3. 8,9.
  4. It is possible to detect viruses in monolayer cells by observing changes in the cells’ shape (e.g., swelling, shrinking, syncytium formation).
  5. When it comes to most viruses, the CPE occurs after 5–10 days of incubation; however, one exception is the herpes simplex virus (HSV), in which the CPE is visible after just 24 hours of incubation.
  6. The kind of virus can be anticipated based on the cell line used for cell culture, the type of specimen, the incubation duration, and the appearance of the CPE; however, confirmation testing, such as an immunofluorescence (IF) assay, is required for a more accurate diagnosis.
  7. The CPE of several viruses in different cell types is depicted in Table 1.
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Table 1

Viruses with distinct CPE formation and confirmation tests were used.

Viruses CPE in


Final identification of isolates
Fibroblasts A549 cells RhMK cells
Adenovirus Some produce clusters Grape-like clusters or “lacy” pattern; 5–8 d Some produce clusters IF for group and neutralization for type
Cytomegalovirus Foci of contiguous rounded cells; 10–30 d CPE
Herpes simplex virus Rounded large cells; 2–6 d Rounded large cells; 1–4 d Some produce CPE IF for group and neutralization for type
Influenza virus Undifferentiated CPE, cellular granulation; 4–8 d IF for group and neutralization for type
Rhinovirus Degeneration, rounding; 7–10 d CPE

IF staining, on the other hand, does not allow for the detection of all viruses. The enterovirus family contains a large number of different serotypes, and IF staining does not distinguish between them all. Aside from that, the sensitivity of the monoclonal antibody utilized for the identification of enteroviruses is low, and there have been reports of cross-reactions between the monoclonal antibody and enteroviral serotypes11,12, and 13. Traditional cell culture methods may be used to identify a wide range of viruses in a variety of cell lines using various cell lines.

While the lengthy incubation and monitoring of the CPE are important drawbacks, the high cost involved from the acquisition and maintenance of various monolayer cells is also a significant drawback.

3. The new format of cell culture

Rather of the standard screw-cap cell culture tube (16 mm x 125 mm), a 1-dram vial or a shell vial is now used instead, which are both smaller (Figure 3). It is possible to develop monolayer cells at the bottom of the vial with the help of this container. Moreover, this procedure makes it simple to centrifuge the results of the experiment.

3.1. Cryopreserved cell culture

Various types of cells are grown as a monolayer in a vial using this procedure, and various monoclonal antibodies are given to these cells in order to diagnose different viruses. It is possible to identify many viruses in the same vial using this approach. The MRC-5 and A549 cell lines were employed as monolayers in a vial for the diagnosis of CMV, herpes simplex virus (HSV), and adenoviruses, respectively. Staining was accomplished by the use of a cocktail antibody. After that, a secondary antibody that had been tagged with antispecies antibodies was added.

  1. The cells were stained with FITC and then analyzed using ultraviolet (UV) filters.
  2. In addition to R-Mix cells, there are several more cell lines that are employed for isolating a range of viral respiratory infections.
  3. Each sample contains three R-Mix cell lines, which are employed in the experiment.
  4. Several different kinds of fluorescein-labeled monoclonal antibodies against adenoviruses, parainfluenza virus types 1, 2, and 3, influenza virus types A and B, respiratory syncytial virus, and influenza virus Type A were administered to R-Mix after 24 hours.

3.3. Virus identification in transgenic cell lines

Transgenic cell lines are used in this procedure, which is both quick and accurate. Some genetic elements are incorporated in the cells, and it is possible to recognize a specific virus utilizing these elements. These substances can be produced from any living organism. In a prior investigation, a CD4-positive lymphoid cell line converted into a retroviral vector with a long terminal repeat promoter in conjunction with the chloramphenicol acetyltransferase gene and HeLa was used to identify the presence of human immunodeficiency virus (HIV).

4. Conclusion

A large number of novel cell culture methods have been proposed since the discovery of the technique (e.g., use of shell vial, cryopreservation). Furthermore, the time necessary for viral detection was significantly reduced, going from 5–10 days (using standard approaches) to 24 hours (using innovative methods) (novel methods). A reduction in the number of distinct cell lines employed in laboratories for viral detection was achieved by combining diverse cells into a single vial.

Furthermore, the innovative cell culture methods are more precise and sensitive when it comes to the identification of viruses. However, it is necessary to add some more precise studies, such as molecular approaches in cell culture, in order to ensure the correct identification of viral strains.

Conflicts of interest

There are no conflicts of interest declared by the author.

References

R.I. Freshney’s Fourth Edition, Wiley-Liss, New York (NY), 2000. 2. pages. 200–204 in Culture of Animal Cells: A Manual of Fundamental Technique. The evolution of diagnostic virology from animals to automation, by G.D. Hsiung. Yale Biol Med 1984; 57(5):727–733. J Biol Med 1984; 57(5):727–733. • Studies on the culture of the vaccinia virus by Stinehardt, Israeli, and Lambert (Eds.). Journal of Infectious Diseases, Volume 13, Number 2, Pages 204–300, 1913. The modification of the virulence of yellow fever virus by culture in tissues in vitro was discovered by Lloyd W., Theiler M., and Ricci N.I.

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In this paper, we describe the use of cryopreserved cell monolayers for the fast detection of herpes simplex virus (HSV) and influenza virus (IAV).

Journal of Clinical Microbiology, 2002; 40 (11):4301–4303.

Three-color fluorescence microscopy was used to demonstrate simultaneous cultivation of adenovirus, CMV, and herpes simplex virus in the same shell vial.

J Clin Microbiol, vol. 32, no. 9, pp. 2289–2290, 1994. 16. P.D. Olivo’s transgenic cell lines for the detection of animal viruses are described. Clinical Microbiology Review, vol. 9, no. 3, 1996, pp. 321–334.

Viral culture – Wikipedia

Viral culture is a laboratory procedure in which samples of a virus are introduced into different cell lines and the virus’s potential to infect them is assessed. If the cells undergo alterations, which are referred to as ascytopathic effects, the culture is considered positive. In most cases, traditional viral culture has been replaced with shell vial culture, in which the material is spun onto a single layer of cells and the amount of viral growth is determined using antigen detection techniques.

Furthermore, the centrifugation phase in shell vial culture improves the sensitivity of this approach because, following centrifugation, the viral particles in the sample are in close proximity to the cells, increasing the sensitivity of the procedure.

Viruses that can be identified by viral culture include adenoviruses, cytomegaloviruses,enteroviruses, herpes simplex virus, influenza virus, parainfluenza virus, rhinovirus, respiratory syncytial virus, varicella-zoster virus, measles, and mumps, among others.

Generally, the ultimate identification method for these viruses is immunofluorescence, with the exception of cytomegaloviruses and rhinoviruses, which are identified in a viral culture by their cytotoxic effects.

See also

  • In this section, you will learn about cell culture, microbiological instruments, laboratory detection of viral infections, viral plaque, and viral illness testing.

References

  1. Paula Tennant and Gustavo Fermin are the authors of this work (2018). “Viruses as Potential Biotechnology Targets.” Aspects of Viruses, 317–338. doi: 10.1016/B978-0-12-811257-1.00013-9.ISBN9780128112571
  2. Curtis, Jeanette
  3. Rea, Caroline
  4. ISBN9780128112571 (25 May 2007). “Viral culture” refers to the spread of information over the internet. WebMD. 9th of September, 2009
  5. Retrieved 9th of September, 2009
  6. Gregory A. Storch, Bernard N. Fields, David Mahan Knipe, and Peter M. Howley are among those who have contributed to this work (2007). “Diagnostic virology” is an abbreviation. The authors David Mahan Knipe and Peter M. Howley have written a book called (ed.). Fields’ Virology, 1st ed (5th ed.). p. 3177. ISBN 978-0-7817-6060-7
  7. AbTable 2 in: Leland DS, Ginocchio CC. Lippincott WilliamsWilkins
  8. P. 3177. ISBN 978-0-7817-6060-7
  9. (January 2007). A role for cell culture in viral detection in the era of technology is discussed in this paper. Journal of Clinical Microbiology, Volume 20, Number 1, pages 49–78. doi: 10.1128/CMR.00002-06.PMC1797634.PMID17223623
  10. Jefferson, Tom
  11. Spencer, Elizabeth
  12. Brassey, Jon
  13. Heneghan, Carl
  14. Jefferson, Tom, Elizabeth Spencer, Jon Brassey, Jon Heneghan, Carl (3 September 2020). “Viral cultures were used to test the infectivity of COVID-19. Review of the literature on a systematic basis “. medRxiv: 2020.08.04.20167932
  15. Doi: 10.1101/2020.08.04.20167932.S2CID220962177
  16. MedRxiv: 2020.08.04.20167932

External links

  • “Viral Culture in General.”LabCorp. “Viral Culture in Specific.” The original version of this article was published on July 13, 2011. “Growing viruses from clinical samples,” which was retrieved on September 9, 2009. The London Laboratory Services Group is a private company based in London. 9 September 2009
  • Retrieved 9 September 2009

9.4B: Tissue Culture of Animal Viruses

Instead of growing viruses in ordinary microbiological broth or on agar plates, viruses must be cultivated within host cells that are compatible with their growth. Objectives for Learning

  • Learn about the benefits of cultivating animal viruses in host cells as well as the reasons for doing so.

Key Points

  • In order to cultivate clinical samples that are suspected of carrying a virus, tissue culture is a valuable technique. This approach is useful in the laboratory for the detection, identification, and characterisation of viruses
  • It is also useful in the field. Growing animal cells in flasks using different broth medium and subsequently infecting these cells with virus is the process of tissue culture of animal viruses. Calcium phosphate, electroporation, or combining a cationic lipid with the material to generate liposomes, which fuse with the cell membrane and deposit their payload within, are all methods for carrying out transfection. When viruses infect cells, they induce non-lytic damage, which is known as cytotoxicity. These present themselves in a variety of ways and have a variety of negative consequences.

Key Terms

  • In cell culture, a sophisticated process through which cells are cultivated under controlled circumstances, usually outside of their natural habitat, is called cell culture. The term “cytopathic effect” refers to the degenerative changes that occur in cells, particularly in tissue culture, and is sometimes connected with the replication of some viruses.

Generally speaking, cell culture is a complicated process in which cells are cultivated under controlled circumstances, usually outside of their native habitat. In practice, the word “cell culture” now refers to the cultivation of cells originating from multicellular eukaryotes, particularly animal cells, rather than the cultivation of single cells. There are, however, cultures of plants, fungi, and microorganisms, including viruses, bacteria, and protists, that have been created. It is important to note that the historical history and current methods of cell culture are intimately tied to those of tissue and organ culture.

  1. Viruses are intracellular parasites that require the presence of live cells in order to reproduce and spread.
  2. Despite the fact that embroyonated eggs and laboratory animals are extremely beneficial for the isolation of specific viruses, cell cultures are the only approach used in the majority of laboratories for viral isolation.
  3. In order to produce cell cultures, tissue pieces must first be dissociated, which is commonly accomplished with the use of trypsin or collagenase enzymes.
  4. Examples include Eagle’s and an animal serum.
  5. The attachment of normal cells to a stable support is required for their growth to be successful.
  6. The sensitivity of various cell cultures to different viruses varies significantly.
  7. As soon as feasible after collection, samples for cell culture should be delivered to the laboratory for further processing.
  8. Fluids and tissues from the body should be collected and stored in a sterile container.
  9. Change the maintenance media after one hour or the next morning, whichever comes first.
  10. Rotation is the most effective method of isolating respiratory viruses, and it also results in the manifestation of cytopathic effects (CPE) for many viruses occurring early in the isolation process.

If stationary tubes are utilized, it is vital that the culture tubes are positioned in such a way that the cell monolayer is bathed in nutritious media. If this is not done, the cells will die.

Practice Questions – Principles of Virology

  1. Cultivation of cells is a complicated process in which cells are cultivated under controlled circumstances, usually outside of their native habitat. It is now well understood that the word “cell culture” is referring to the process of growing and cultivating cells that have been obtained from multicellular eukaryotes, particularly animal cells. There are, however, cultures of plants, fungi, and microorganisms, including viruses, bacteria, and protists, that may be found in the laboratory. It is important to note that the historical history and current methods of cell culture are intimately linked to those of tissue and organ culture. Despite the fact that animal cell culture became a widely used laboratory method in the mid-1900s, the notion of retaining living cell lines that were isolated from their original tissue source was found in the 19th century. Viruses are intracellular parasites that require the presence of live cells in order to reproduce successfully. It is possible to isolate viruses from cultured cells, eggs, and laboratory animals. Cell cultures are the only technique used in most laboratories for viral isolation, despite the fact that embroyonated eggs and laboratory animals are quite beneficial for specific viruses. In order for animal virology to proceed, it is necessary to create ways for producing animal cells in culture. Typically, trypsin or collagenase are used to separate tissue pieces prior to culturing them in cell culture. It is then placed in a flat-bottomed glass or plastic container (a petri dish, a flask, a bottle, or test tube) and mixed with a liquid medium that is appropriate for the cell suspension. Example: an animal serum such as Eagle’s and a fish serum It is possible that the cells may connect and spread on the bottom of the container after a variable lag time and then begin to divide, resulting in the development of a primary culture. Normal cells must be attached to a strong support in order to develop properly. Figure: Herpes simplex virus has a cytotoxic impact, which is a kind of cytotoxicity. The sensitivity of various cell cultures to different viruses varies substantially from one to the other. If a suspected virus is detected in a cell culture, it is critical that the most sensitive cell culture be employed. As soon as feasible after collection, samples for cell culture should be delivered to the laboratory for further analysis. A vial containing viral transport medium should be placed in which the swabs will be placed. It is necessary to put bodily fluids and tissues in a sterile container. When the material is received, it is injected into a variety of various types of cell culture, which are selected based on its nature and clinical presentation. After one hour, or the next morning, the maintenance medium should be replaced. Incubate the infected tubes at 35-37 degrees Celsius in a revolving drum for at least 24 hours. When it comes to the isolation of respiratory viruses, rotation is the most effective method. It also results in the manifestation of cytopathic effects (CPE) for many viruses occurring earlier than would otherwise be the case. For cell monolayers to be bathed in nutritional medium, it is vital that the culture tubes are positioned so that the cell monolayer is immersed in the nutrient media.
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When cells are cultivated under controlled settings, often outside of their natural habitat, they are said to be in cell culture. Today, in practice, the phrase “cell culture” refers to the process of cultivating cells generated from multicellular eukaryotes, most notably animal cells, for use in research. There are, however, cultures of plants, fungi, and microorganisms, including viruses, bacteria, and protists, that are present. It is important to note that the historical history and current methods of cell culture are intimately linked to those of tissue culture and organ culture.

  • Viruses are intracellular parasites that require the presence of live cells in order to reproduce.
  • Despite the fact that embroyonated eggs and laboratory animals are extremely beneficial for the isolation of specific viruses, cell cultures are the only approach used in most laboratories for viral isolation.
  • In order to produce cell cultures, tissue pieces must first be dissociated, which is commonly accomplished with the use of trypsin or collagenase.
  • For example, Eagle’s and an animal serum.
  • The attachment of normal cells to a solid support is required for their growth.
  • The sensitivity of various cell cultures to different viruses varies substantially.
  • It is important to bring specimens for cell culture to the laboratory as soon as they are collected.
  • Fluids and tissues from the body should be deposited in a sterile container.
  • After one hour, or the next morning, the maintenance medium should be replaced.
  • Rotation is the most effective method of isolating respiratory viruses, and it also results in the manifestation of cytopathic effects (CPE) in many viruses occurring earlier than would otherwise be the case.

If stationary tubes are utilized, it is vital that the culture tubes are positioned in such a way that the cell monolayer is bathed in nutritious liquid.

  1. Cell culture is a complicated process in which cells are cultivated under controlled circumstances, usually outside of their native environment. In practice, the word “cell culture” now refers to the cultivation of cells obtained from multicellular eukaryotes, particularly animal cells. There are, however, cultures of plants, fungi, and microorganisms, including viruses, bacteria, and protists, that can be found. The historical evolution and methods of cell culture are intricately intertwined with those of tissue culture and organ culture. Animal cell culture became a widely used laboratory technique in the mid-1900s, however the notion of keeping living cell lines that were isolated from their original tissue source was found in the nineteenth century. Viruses are intracellular parasites that require live cells in order to reproduce. For viral isolation, cultured cells, eggs, and laboratory animals may be employed. Although embroyonated eggs and laboratory animals are extremely effective for the isolation of specific viruses, cell cultures are the only approach used in the majority of laboratories for viral isolation. The invention of ways for culturing animal cells has been critical to the advancement of animal virology. To produce cell cultures, tissue pieces must first be dissociated, which is commonly accomplished with the use of trypsin or collagenase. The cell suspension is then put in a flat-bottomed glass or plastic container (e.g., a petri dish, a flask, a bottle, or a test tube) along with an appropriate liquid medium. For example, Eagle’s and an animal serum are two examples. After a variable amount of time, the cells will adhere to and spread on the bottom of the container, and then begin to divide, resulting in the formation of a primary culture. The attachment of normal cells to a stable substrate is necessary for their development. Figure: Herpes simplex virus has a cytotoxic effect on the body. The sensitivity of various cell cultures to different viruses varies widely. When testing for a suspected virus, it is critical to employ the most sensitive cell cultures possible. Specimens for cell culture should be brought to the laboratory as soon as feasible after they are collected. Swabs should be placed in a vial containing viral transport medium. Body fluids and tissues should be collected and stored in a sterile container. Upon reception, the specimen is injected into one of many distinct types of cell culture, depending on the nature of the material and the clinical presentation. The maintenance medium should be replaced after one hour or the next morning, whichever comes first. The inoculation tubes should be incubated at 35-37 degrees Celsius in a revolving drum. Rotation is the most effective method of isolating respiratory viruses, and it also results in the manifestation of cytopathic effects (CPE) for many viruses occurring earlier than would otherwise be the case. If stationary tubes are employed, it is vital that the culture tubes be positioned in such a way that the cell monolayer is bathed in nutritious liquid.

What mechanism does a virus employ to move chemicals through its membrane?

  1. Active transport, facilititated diffusion, endocytosis, pinnocytosis, group translocation, simple diffusion, all of the above, none of the above, none of the above

Viruses may be disassembled (split) into two parts: the capsid proteins and the nucleic acid. After being reassembled, these two pieces will self-assemble into new infectious virus particles that will spread throughout the body. You will purify the NUCLEIC ACID from the TOMATO mosaic virus and the PROTEIN from the BEAN mosaic virus from their respective viruses. These two pieces are then combined, and they self-assemble to form viruses that are harmful to humans and animals. When these freshly constructed (hybrid) viruses are rubbed (with an abrasive) into BEAN LEAVES, which one of the following should happen?

  1. The virus will infect bean leaves and cause tomato mosaic viruses to be produced. Bean leaves will be infected by the virus, which will then create bean mosaic viruses. The virus will infect bean leaves and create both bean mosaic virus and tomato mosaic virus as a result of infection. Because the virus will not infect bean leaves, no viruses will be created. Bean leaves will be infected by a virus, but lysogeny will hinder the formation of new viruses.

The primary function of the early viral genes that are expressed is to infect cells.

  1. Take over the host machinery
  2. Synthesis nucleic acids
  3. Uncoating the virus
  4. Causing sickness
  5. Making the protein coat
  6. None of these things
  7. None of these things.
  1. They can only reproduce in particular types of cells
  2. Hence, they are restricted. Only certain types of cells are susceptible to viral infections
  3. The others are not. They can only be absorbed by cells that have the appropriate receptors. They can only penetrate cells that contain glycoproteins. Cells with the opposite charge will only link to them
  4. Otherwise, they will not bind.

All of the following explanations of viral multiplication and nucleic acids are accurate, with the exception of the following:

  1. Viruses possess either DNA or RNA, not both
  2. They can only reproduce in live cells
  3. They exploit the cell’s biosynthetic machinery to create copies of themselves
  4. And they are contagious. When a virus replicates, its nucleic acid is protected by a protein coat
  5. Viralm RNA, viralt RNA, and viral ribosomes are all components of viral reproduction.

Only live cells can multiply viruses; viruses employ the cell’s biosynthetic machinery to create copies of themselves; viruses do not contain DNA or RNA; viruses do not replicate in the absence of living cells; viruses do not replicate in the absence of living cells When a virus replicates, its nucleic acid is protected by a protein coat; viralm RNA, viralt RNA, and viral ribosomes are all involved in the process.

  1. Only living cells can replicate viruses
  2. Viruses use the cell’s biosynthetic machinery to synthesize copies of themselves
  3. Viruses do not contain DNA or RNA
  4. Viruses do not replicate in the absence of living cells
  5. Viruses do not replicate in the absence of living cells. When a virus replicates, its nucleic acid is protected by a protein coat
  6. Viralm RNA, viralt RNA, and viral ribosomes are all involved in the process.

Viruses have either DNA or RNA, but not both; viruses can only reproduce in live cells; viruses employ the cell’s biosynthetic machinery to create copies of themselves; viruses are not contagious. The nucleic acid of a virus is enveloped by a protein coat; viralm RNA, viralt RNA, and viral ribosomes are all components in viral replication;

  1. Viruses possess either DNA or RNA, not both
  2. They can only reproduce in live cells
  3. They exploit the cell’s biosynthetic machinery to create copies of themselves
  4. And they are contagious. When a virus replicates, its nucleic acid is protected by a protein coat
  5. Viralm RNA, viralt RNA, and viral ribosomes are all components of viral reproduction.

All of the following claims are correct in terms of the virion’s attachment or adsorption to the host cell, with the exception of the following:

  1. It is the surface capsid proteins of naked viruses that cause them to attach to a specific cell receptor
  2. For enveloped viruses, it is the spikes of the virus that are responsible for binding to a specific cell receptor
  3. Thus, a cell that lacks a specific viral receptor is not infected by that virus. It is true that animal viruses, such as influenza and HIV, are extremely specific in that they can only associate with specific cell types
  4. Plant viruses, such as tobacco mosaic virus, require specific receptor sites in order to attach to cells
  5. And that all of the above statements are correct

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