Test 2: Dx, Tx, and Prevention Flashcards

1
Q

What does PCR stand for?

A

Polymerase Chain Reaction

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2
Q

What is PCR used for?

A

PCR is used to make

multiple copies of a segment of DNA

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3
Q

What is it called when you take one segment of DNA and make multiple copies of it?

A

Amplification

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4
Q

The following steps describe this process of diagnosing viral infections:

  1. Amplify DNA
  2. Subject DNA to electrophoresis in agarose gels
  3. Stain with ethidium bromide
A

PCR

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5
Q

In PCR, what temperature is used to denature (or separate) the strands of DNA?

A

94 - 96C

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6
Q

In PCR, what temperature is used to **anneal **the primers to the ends of both strands of denatured DNA

A

68C

for annealing

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7
Q

T/F:

In the annealing step of PCR,

the forward primer (+) binds to the forward DNA strand (+)

A

FALSE!!!

the forward primer (+)

binds to the

reverse DNA strand (-)

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8
Q

In PCR, what temperature is used to

elongate each strand of DNA after they have been annealed to their primers?

A

72C

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9
Q

The **elongation **step in PCR uses

______ polymerase to create a new DNA segment that is

complementary to the original DNA strand

A

TAQ polymerase

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10
Q

What is the result of the **elongation **step of PCR?

A

2 new DNA molecules

each with

1 **NEW **strand

and

1 OLD strand

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11
Q

Repeating the process of PCR for 30 - 40 cycles

yields how many exact copies of the original DNA segment?

A

1 billion!!!

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12
Q

While PCR is used to make multiple copies of DNA,

what is QPCR used for?

A

Quantitative PCR

is used for the

  • monitoring and quantification*
  • of PCR products and nucleic acid load*
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13
Q

Why is QPCR useful?

A

QPCR is used to monitor the

viral load in a patient

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14
Q

What 3 target-specific probes are used in the **QPCR **process?

A
  1. 5’ TaqMan probe
  2. Molecular Beacon probe
  3. FRET hybridization probe

Taqman frets when he can’t find bacon, so he probes for it!

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15
Q

Intercalating dyes, like SYBR green are used in what diagnosis technique?

A

QPCR

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16
Q

What is a flurophore?

(SYBR green for example)

A

Flurophore is a probe or a dye that emits fluorescence

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17
Q

What is the most widely used method of

Genome Sequencing?

A

Sanger Dideoxy Method

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18
Q

What does NGS stand for?

A

New Generation Sequencing

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19
Q

Illumina (Solexa) sequencing is an example of this type of genome sequencing

A

NGS

new generation sequencing

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20
Q

Which method of genome sequencing is

cheaper

quicker

more accurate and reliable

and

Needs LESS DNA?

A

Illumina (Solexa) new generation sequencing

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21
Q

The study of microbial populations in a sample by

analyzing the sample’s entire nucleotide sequence

is called?

A

Metagenomics

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22
Q

What is metagenomics used for?

A

The random detection of existing and new pathogens

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23
Q

NGS platforms are being used for the formation of

___________ databases

A

metagenomics

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24
Q

Genome sequencing is crucial in

_______ studies for pathogen detection using metagenomics

A

surveillance

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25
Q

The use of virus genome sequence data to study

evolution and genetic relationships of viruses

is called

A

phylogenetic analysis

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26
Q

Phylogenetic analysis has been used to related

porcine rotavirus to human rotavirus.

Which lineage is the porcine one and which is the human one?

A

Lineage 1 is porcine (RU172/G12)

Lineage 2 is human (RVA/G12)

both are related to the VP7/G12 genotype

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27
Q

T/F:

QPCR is the same thing as RTPCR

A

TRUE!

Quantitative PCR is the same as Real Time PCR

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28
Q

Which process of viral detection can screen hundreds of pathogens simultaneously and uses only 1 silicon chip?

A

Microarrays

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29
Q

In a microarray

**probe DNA **and sample DNA interaction

is called

A

hybridization

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30
Q

What does a positive reaction in a microarray test for pathogens look like?

A

Fluorescence on the chip where the probe was

is indicative of a (+) reaction

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31
Q

What are the 4 ways of treating viral diseases?

A
  1. Antivirals
  2. Immune system stimulation
  3. Antibody synthesis
  4. Antibody administration (natural antiserum)
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32
Q

This class of proteins with antiviral effects is used to stimulate the immune system in the treatment of viral diseases

A

interferons

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33
Q

T/F:

You can use an antibiotic to treat a viral infection

A

FALSE!!

Antibiotics should be known as “antibacterials” because they only treat BACTERIAL infections

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34
Q

Compared to antibiotics, only a few antivirals are effective. Why is this?

A

Agents that kill viruses will also kill the host cell

(because viruses depend on the host cell for replication)

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35
Q

Antiviral chemotherapeutics are NOT common in vet med. Why not?

A

The cost is too high

It’s hard to find effective ones that are not cytotoxic

There are no rapid diagnosis tests so acute-onset viral infections cannot use them anyway

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36
Q

What are the 6 steps in the Virus life cycle?

A
  1. Receptor binding
  2. Cell entry
  3. Uncoating
  4. Nucleic acid synthesis
  5. Assembly
  6. Release
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37
Q

What are **receptor analogs (antiviral drug) **used to target?

A

Receptor analogs target

*attachment *of the virion to the cell receptor

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38
Q

What Rimantadine (antiviral drug) used to target?

A

Rimantidine targets

uncoating

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39
Q

What are transcriptase inhibitors (antiviral drug) used to target?

A

Transcriptase inhibitors target

primary transcription from viral genome

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40
Q

What antiviral drug targets

reverse transcription?

A

Zidovudine (AZT)

targets viral reverse transcription

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41
Q

**Lentivirus TAT inhibitors **are antiviral drugs used to target what?

A

Lentiviral TAT inhibitors

target

regulation of transcription

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42
Q

What is Ribavirin (antiviral drug) used to target?

A

Ribavirin

targets the

processing of RNA transcripts

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43
Q

Interferons, when used as antiviral drugs, target

A

translation of viral RNA into proteins

is the target of Interferons

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44
Q

Post-translational cleavage of proteins is the target of this type of antiviral drug

A

Protease inhibitors

45
Q

What is Acycloguanosine (Acyclovir) (antiviral drug) used to target?

A

**Acyclovir **targets

replication of the viral DNA genome

46
Q

Replicase inhibitors are used as antivirals to target what?

A

Replicase inhibitors target

replication of viral RNA genome

47
Q

**Acyclovir **and Replicase inhibitors

are both used to target the **replication **step of viral genomes.

Which one targets the viral DNA genome?

A

Acyclovir

targets viral DNA genomes

48
Q

**Acyclovir **and **Replicase inhibitors **

are both used to target the replication step of viral genomes.

Which one targets the viral RNA genome?

A

Replicase inhibitors

target viral RNA genomes

49
Q

What antiviral is used to treat

Herpesvirus

by inhibiting the replication of viral DNA genome?

A

Acyclovir

is used to treat

Herpesvirus

50
Q

Acyclovir is used to treat

corneal ulcers in cats and encephalomyelitis in horses

afflicted with what virus?

A

Herpesvirus-1

51
Q

In the mechanism of treating Herpesvirus using Acyclovir,

Herpesvirus _______ kinase

changes Acyclovir into Acyclovir monophosphate

A

Thymidine Kinase

converts

Acyclovir –> Acyclovir monophosphate

52
Q

In the mechanism of treating Herpesvirus using Acyclovir,

_______ kinase

changes Acyclovir monophosphate into Acyclovir diphosphate

A

Cellular (GMPK) Kinase

Acyclovir monophosphate –> Acyclovir diphosphate

53
Q

In the mechanism of treating Herpesvirus using Acyclovir,

_______ kinase

changes Acyclovir *diphosphate into Acyclovir *triphosphate

A

Cellular (NDPK) Kinase

Acyclovir *diphosphate –> Acyclovir *triphosphate

54
Q

Herpesvirus DNA polymerase cleaves acyclovir triphospate into

acyclovir __________

A

Acylovir Monophosphate

by using Herpesvirus DNA polymerase

55
Q

What happens when **acyclovir monophosphate **is incorporated

into the growing DNA strand?

A

Further elongation of the DNA is impossible and

DNA chain replication of the virus STOPS!

56
Q

**Acyclovir monophosphate **stops replication of viral DNA

when incorporated into the growing DNA strand

because it lacks _________

A

Acyclovir monophosphate lacks

an attachment point for nucleotides

57
Q

In the treatment of Herpesvirus by Acyclovir

Acyclovir monophosphate is what stops replication of the virus.

Acyclovir triphosphate also helps in fighting the virus. How?

A

Acyclovir triphosphate

is a competitive inhibitor

of dGTP for viral DNA polymerase

58
Q

Acyclovir triphosphate

is a competitive inhibitor

of _____ for viral DNA _______

A

dGTP

for

viral DNA polymerase

59
Q

T/F:

Acyclovir can be cytotoxic

A

FALSE!

Acyclovir is NOT harmful to the infected host cell

60
Q

**TK negative, partial, **and altered mutants

are resistant to treatment of ___________

by __________

A

Herpesvirus

by

Acyclovir

61
Q

Alterations of viral DNA polymerase

result in decreased _________ of

Acyclovir __________ to viral DNA polymerase

A

decreased binding

of

Acyclovir triphosphate to viral DNA polymerase

62
Q

What type of Herpesvirus mutant is substate specificity altered

and results in phosphorylation of thymidine, but NOT Acyclovir?

A

TK altered mutants

63
Q

Amantidine is used to treat which 2 diseases

by inhbiting replication by blocking UNCOATING?

A

Amantidine (RImantidine)

treats

Influenza A and Parkinson’s Disease

64
Q

In the process of Influenza virus replication,

when the pH in the endosome containing the virus drops to _____,

HA protein has a change in conformation

allowing viral RNA to be released into the cytoplasm of the host and replicate

A

pH of 5.0

65
Q

In the mechanism of infection of Influenza virus,

Viral _______ ion channel pumps protons into the endosome

in order to ____________

which causes the release of viral RNA from the M1 matrix protein

A

In the mechanism of infection of Influenza virus,

Viral M2 ion channel pumps protons into the endosome

in order to lower its pH to 5.0

which causes the release of viral RNA from the M1 matrix protein

66
Q

Amantidine blocks uncoating in influenza virus infections

by clogging the M2 channel which prevents what?

A

**Amantidine **clogs the M2 channel

which prevents

protons from being pumped into the endosome to lower the pH

RNA remains bound to M1 matrix protein and viruses replication stops

67
Q

**Amantidine **is prevented from clogging the M2 channel when

there are changes in the ______ _______ that line the M2 channel

A

amino** acids**

68
Q

What are Neuramidase Inhibitors used to treat?

A

Influenza virus

69
Q

What is the function of neuramidase (NA)?

A

NA cleaves sialic acid receptors on the cell surface which releases HA

which is critically important in the spread of Influenza virus

70
Q

T/F:

Neuramidase inhibitors kill off influenza virus in the host.

A

FALSE

Neuramidase inhibitors slow the spread of influenza virus and allows the immune system to “catch up”

71
Q

Peramivir

Zanamivir

Oseltamivir

Laninamivir

are all __________ ____________

A

Neuramidase Inhibitors

used to treat Influenza

72
Q

Which 5 drugs are used to treat influenza virus?

4 are neuramidase inhibitors. Which one is not?

A
  1. Peramivir
  2. Zanamivir
  3. Oseltamivir
  4. Laninamivir
  5. Amantidine

Amantidine blocks influenza viral uncoating, not NA

73
Q

What are the 4 targets of Anti-retroviral therapy?

A
  1. Fusion
  2. Integrase
  3. Reverse Transciptase
  4. Protease
74
Q

There are 4 methods of preventing the spread of antiviruses:

Drugs that inhibit fusion, integrase, reverse transcriptase, and protease.

Which drugs are reverse transcriptase inhibitors aka NRTIs?

A

Reverse Transcriptase Inhibitors (NRTIs):

Zidovudine

Azidothymidine

(ZDV or AZT)

75
Q

There are 4 methods of preventing the spread of antiviruses:

Drugs that inhibit fusion, integrase, reverse transcriptase, and protease.

Which drugs are HIV protease inhibitors?

A

HIV Protease Inhibitors:

Saquinavir

Idinavir

Nelfinavir

Ritonavir

(SINR)

76
Q

ZDV and AZT are NRTIs

What does “NRTI” stand for?

A

Nucleoside-analog Reverse Transcriptase Inhbitors

77
Q

ZDV and AZT are nucleoside-analogs of __________

a nucleobase in DNA

A

thymine

78
Q

How does ZDV and AZT stop the replication of HIV?

A

ZDV and AZT have an azide group (N3) instead of OH (hydroxyl)

which has nowhere for DNA to elongate

79
Q

45% of animals experience major toxicities when taking ________

like anemia and granulocytopenia

A

AZT or ZDV for HIV

80
Q

How do protease inhibitors like Saquinivir, Indiavir, Nefeltavir, and Ritonavir

treat HIV?

A

Proteases are required to cleave HIV polyproteins into functional proteins!!

Protease inhibitors stop the cleaving and HIV cannot mature, thus

non-infectious viruses are produced

81
Q

What are the 4 W’s of Immunization?

A
  1. Where
    1. Endemic area populations
  2. When
    1. Right before seasonal viruses hit
    2. If an outbreak of a non-endemic virus occurs
  3. Who
    1. Populations at-risk
  4. Why
    1. The loss caused by disease must be greater than the cost of immunity
82
Q

What is an attenuated-virus?

A

A viral mutant that no longer has virulent capabilities, does not cause disease

83
Q

What type of vaccine is formed though

*serial dilulations of virulent viruses in cultured cells *

with the hopes that it will

*acquire and accumulate nucleotide substitutions *

resulting in avirulence?

A

Live-Attenuated Vaccines

84
Q

Cold adapted mutant virus Live-Attenuated Vaccines

are safer for __________ administration

(Equine Influenza treatment)

A

intranasal

85
Q

A vaccine produced from purified native viral proteins

is called a _______ vaccine, a type of

Non-replicating Virus Vaccine

A

SPLIT vaccine

is a type of non-replicating vaccine

86
Q

A vaccine produced from inactivated whole virions is called an

Inactivated Virus Vaccine

which is a type of ___________ vaccine

A

Non-replicating Virus Vaccine

87
Q

Inactivated viruses are Non-replicating viruses made from

virulent viruses using ________ or ___________ agents

to destroy infectivity while maintaining immunogenicity

A

Chemical or Physical agents

88
Q

Split Vaccines (purified native viral proteins) are Non-Replicating Virus Vaccines

produced from solubilized virions using

________ _________ to semi-purify the glycoproteins

A

differential centrifugation

89
Q

Some vaccines are produced using recombinant DNA and technology

These include vaccines produced by

Attentuation, Sub-unit markers, microbial vector vaccines and _________ vaccines

A

DNA vaccines

90
Q

Recombinant DNA and Technology Produced Vaccines

include viruses attenuated using what 2 methods?

A

Gene deletion

Site-directed mutagenesis

91
Q

This type of vaccine produced by recombinant DNA and technology

is produced by expression of viral proteins in

eukaryotic, bacterial, and plant cells

A

Subunit “marker” vaccines

92
Q

This type of vaccine produced using recombinant DNA and technology

utilizes harmless viruses as vectors of expression for other heterologous viral antigens

and integrates **viral HA **into the viral vector

A

Microbial Vector Vaccines

93
Q

This type of vaccine produced using recombinant DNA and technology

utilizes viral DNA and integrates it into a plasmid

A

DNA vaccines

94
Q

It is impossible to differentiate an animal vaccinated with ____________ vaccines

from animals infected by a field strain

A

Live-attenuated viruses

95
Q

Subunit “Marker” Vaccines

have _____ antigens than natural field strains

A

less antigens

96
Q

What are the 8 methods of controlling viral vectors?

A
  1. Source Reduction
  2. Biological Control
  3. Chemical Control
  4. Contact Potential Reduction
  5. Portals of Entry Protection
  6. Decontamination
  7. Surveillance
  8. Farm Biosecurity
97
Q

The process that destroys

all forms of microbial life and pathogens including bacterial spores

is called

A

Sterilization

98
Q

What are the 5 methods of Sterilization

that kill all pathogens including bacterial spores?

A
  1. Steam Autoclave
    1. 121C for 15 mins at 15 psi
  2. Dry Heat
    1. 160C for 2 hours
  3. Chemicals
    1. Ethylene oxide or Ozone gas
    2. High concentration hydrogen peroxide
  4. Radiation
    1. Non-ionizing UV radiation
    2. Ionizing Gamma or X-rays
  5. Sterile Filtration
    1. Microfiltration with membrane filters with pores 0.2µm
99
Q

The process of decontamination that

destroys most microbes and pathogens except bacterial spores

is called

A

Disinfection

100
Q

The decontamination process of

applying a liquid antimicrobial chemical to skin to destroy microorganisms

is called

A

Antisepsis

101
Q

Sentinel animals serve as *early warning systems *for _________ diseases

A

ZOONOTIC

102
Q

Which sentinel animal is used as an zoonotic early warning system for

West Nile Virus

and

Equine Encephalitis?

A

CHICKENS!

103
Q

Which sentinel animal is used as a zoonotic early warning system for

Rift Valley Fever?

A

Goats

104
Q

Which sentinel animal is used as a zoonotic early warning system for

Lyme Disease?

A

DOGS!

105
Q

Which sentinel animal is used as a zoonotic early warning system for

Yellow Fever?

A

Monkeys :D

106
Q

Which two systems can be used to maintain farm biosecurity and help control viral vectors?

A

All-In or All-Out System

Closed Herd System

107
Q

The quarantine period depends on the _________ period of important infectious diseases

A

incubation period

108
Q

Which levels of biosecurity are concerned with exposure to zoonotic diseases

A

High Risk Level 3

and

Very High Risk Level 4