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1
Q

Immunotherapy tests.

A

Detect pathogen -specific antibody or antigen

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2
Q

Antigens from pathogens can be

A

Whole pathogen
Molecule produced by pathogen
Pathogen molecule presented on surface of host

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3
Q

What are the common antibodies detected in blood are are common specimens for antibodies

A

IgG
IgM
(Found at the area of the infection where the pathogen replicates or the antigen is present

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4
Q

What factor is key in accurate immunochemicla diagnosis?

A

Timing

Acute phase detection may have different antibody presence then further along in disease process

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5
Q

What are indicators of active/recent infection?

A

Pathogen detection
Present/ recent clinical symptoms of infection
Amount or title of antibodies (number circulating system)

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6
Q

How does an ELIZA immunological assay work?

A

Antibody is conjugated with an enzyme, the antibody binds to the antigen.

Substrate is added and the enzyme cleaves a reaction to produce a color change (more color means more binding of antigen-antibody binding)

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7
Q

What are the two different ELIZA assays?

A

Indirect - antibody binding to antibody bound by antigen (antigen coated well)

Sandwich - monoclonal antibody coated well binds antigen and a secondary antibody will bind to the antigen

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8
Q

Describe a lateral-flow immunochromatographic assay

A

Sample is added to pad
Flow of sample by capillary action
Plate antibody binds conjugate antigen and antibody Second line of anybody’s is to bind conjugate antibody (control line)

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9
Q

What is the basis for IDEXX SNAP test?

A

Conjugation of antigens to antibody(bound to plate), a congregate antibody(enzyme bound) will bind to antibody- antigen complex.

Color change is produced when substrate is washed across

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10
Q

What is indirect immunofluorescence?

A

Primary antibody is specific to the antigen (variable region). Binds to antigen
Secondary antibody is specific to the FC portion of the primary antibody -> binds to primary antibody
Secondary antibody is conjugated with fluorophore.

Multiple secondary antibodies can bind and amplify signal

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11
Q

What is agglutination

A

Particles clump together (antibody and antigen interaction)

Direct or Indirect

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12
Q

What is indirect agglutination

A

Antigen or antibody coated on beads

Bind bead through an intermediate

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13
Q

What is direct agglutination

A

Antibody binds directly to antigen.

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14
Q

What are the advantages of immunochemical tests?

A

ID pathogen when pathogen cannot be cultured
Most have high sensitivity
Most have high specificity
Mid to high-volume testing possible

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15
Q

What are the disadvantages of immunochemical tests?

A

Detection of antibody may not indicate an active infection
Antibody detection from specimens: very early in infection may not be detected
Possible that antibodies may detect >1 pathogen (different pathogens can have the same antigen)

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16
Q

What is the basis for molecular diagnostics of infectious disease?

A

Identify makers in the genome or proteome

  • determine pathogen ID by characteristic genetic or protein material
  • use pathogen-specific genetic sequences to ID pathogen
  • use pathogen-specific “protein profile” to ID pathogen
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17
Q

What is MALDI-TOF and how does it work?

A

Mass spectrometry
Detect part of pathogen (ribosomal peptides) by mass and charge
Signature pattern of fragments

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18
Q

What are the advantages of MALDIVES-TOF?

A

Rapid ID
High-throughput
ID bacteria/fungi

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19
Q

What are the disadvantages of MALD-TOF

A

Isolated pathogen analysis
Identification is limited to reference spectra in database
High initial cost

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20
Q

Multiplex PCR/ Microarrays are useful to detect?

A

Nucleic acid from virus, bacteria, fungi parasite species

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21
Q

Real-time PCR

A

Pathogen-specific sequence amplification of nucleic acid and measurement
Quantitative for pathogen
Used highly for viral identification and viral load

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22
Q

What are the advantages of molecular diagnostics?

A

Faster than culture based methods
Highly sensitive
Accurate
High volume testing is possible

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23
Q

What are the disadvantages of molecular diagnostics?

A

Expensive: equipment and reagents
Requires specialized personnel to run machines
Yes or no answers
Possible false negatives/positives

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24
Q

What phenotypic method cannot be used to clinically diagnose viruses

A

Culture-based methods

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25
What three things are required for proper specimens collection for diagnostic testing?
Aseptic technique Collection of a sample specific to the infection Collection before antibiotic treatment
26
What are the common specimens collected for a suspected viral infection?
Swabs- nasal, trachea, sputum, and eye Feces Blood Dependent upon suspected viral infection and patient symptoms
27
Specimen handling is dependent on ?
Pathogen type Specimen type What test will be performed
28
All specimens must be handled to ?
Avoid contamination of other specimens Avoid contamination of clinic/workers/patient
29
What are the categories of diagnostics techniques?
Molecular Immunological and Serological Phenotypic
30
What are phenotypic methods of diagnosis
Direct examination Cytology Biochemical tests
31
What are the concentration techniques for direct examination of a sample?
Filtration/ centrifugation Flotation/ sedimentation Baermann test for larval identification
32
Parasite diagnostics are dependent on??
Stage of infection Animal age and species Technique used Severity of infection False negatives can occur *
33
What are the advantages of cytology ?
``` Determine cell and tissue morphology Cellular association of bacteria/ parasites/ fungi Morphology Impression of disease stage/severity Immediate analysis ```
34
Disadvantages of Cytology
Mild/chronic infection may not be readily detected Mot all samples are appropriate for cytology of bacterial infections Difficult to distinguish normal flora from pathogenic bacteria
35
What type of infections can you culture?
Bacteria or fungal
36
What types of infections are not cultured for diagnosis?
Parasites or viruses
37
What are the different culture options?
Agar Broth Biochemical
38
What type of media is used for general growth of a culture?
Nutrient agar
39
What type of medial is important for growth of a single suspected pathogen type?
Selective media
40
What type of media are used to ID a pathogen?
Differential media Most are also selective medias
41
An enrichment broth is used for what?
To increase the number of a specific bacteria type and inhibit growth of others
42
What is a fastidious bacteria?
Requires specific nutrients and culture conditions | Use environmental/nutritional characteristics to select growth of specific organisms => selective media
43
How can biochemical tests be used to ID a pathogen?
Different bacteria produce different enzymes-> enzymatic or fermentation tests to determine differences in bacterial profiles
44
What is an antibiotic?
Chemical substance that is produced by a microorganism and has the ability to inhibit growth/ kill another microorganism
45
What is the difference between an antibiotic and antimicrobial/antibacterial?
Antibiotics are compounds that are produced by microorganisms Antimicrobial/antibacterial can be any substance (natural/synthetic/semi-synthetic) that inhibits or kills and microorganism All antibiotics are antimicrobial but not all antimicrobials are antibiotics
46
What are the uses of antibiotics in animals?
Therapeutic - treat the sick Prophylactic - prevention Metaphylatic -treat sick and healthy in a heard for prevention Growth promotion - healthy animals treated
47
How can antibacterial agents be classified?
``` Chemical structure Origin Effect on bacteria Spectrum of activity Mode of action ```
48
What compounds are penicillins?
Penicillin G/V Ampicillin/amoxicillin Amoxicillin/ Clavulanic acid
49
What compounds are tetracyclines?
Tetracycline Oxytetracycline Doxycycline.
50
What compounds are Quinolones ?
Oxolinic acid Erirofloxacin/ Marofloxacin Pradofloxacin / Norfloxacin
51
What compounds are Sulfonamides ?
Sulfadiazine | Sulfadiazine/ Trimethorprim
52
What compounds are phenicols?
Chloramephenciol | Florfenicol
53
What compounds are cephalosporins?
Cefalexin/Cephadroxil Cefpodoxime/Ceftiofur Cefovecin, Cefquinome
54
What compounds belong to the macrolides?
Erythromycin | Tiamulin, Tilmicosin
55
What compounds belong to the group Lincosamides
Lycomycin/ Clindamycin | Used for skin infections in dogs
56
What compounds belong to animoglycosides?
Streptomycin Gentamicin/ Neomycin Amikacin
57
What compounds belong to the polypeptide group?
Colistin/ Polymixin
58
What drug targets anaerobes?
Metronidazole
59
What compounds belong to the carbapenems?
Imipenem/meropenem | Ertapenem/ doripenem
60
What compounds are glycopeptides ?
Vancomycin/ Teicoplanin
61
What compounds belong to Oxazolidones?
Linezolide
62
What two antibacterials are synthetically made?
Quinolones and sulfonamides
63
What is a semi-synthetic compound?
A natural compound that is chemically altered
64
What is the difference between a bacteriocidal and bacteriostatic drug?
Bactericidal- kill Bacteriostatic- inhibit growth Drugs can vary depending on concentration, bacterial species, and combinations of drugs
65
What classes of antibiotics are bateriostatic?
``` Lincosamides Macrolides Phenicols Sulfonamides Tetracyclines ```
66
What drug will have no effect against a non-penicillase producing staphylococci?
Metronidazole
67
What group of B-lactams will have no effect on penicillase-producing staphylococci?
Penicillins Aminopenicillins Metronidazole
68
What drugs will have no effect on glucose fermentative gram-negative rods?
Metronidazole Penicillins Lincosamides Macrolides
69
What drugs will have no effect against anaerobes ?
Aminoglycosides
70
What drugs have low/variable activity against Non-penicillase-producing gram-positive cocci ?
``` Lincosamides Macrolides Tetracyclines Aminoglycosides Floroquinolones Cephalosporins Sulfonamides ```
71
What drugs have low/poor activity against penicillase-producing staphylococcus?
Linosamides | Macrolides
72
What drugs have low/variable activity against glucose-fermentation gram-negative rods?
Aminopenicillins | Tetracycline
73
What drugs have low/variable activity against anaerobes?
Cephalosporins Tetracyclines Sulfonamides Flouroquinolones
74
What modes of action can B-lactams have against bacteria?
Inhibit cell wall synthesis, protein synthesis, DNA synthesis, metabolic processes, and cell membrane integrity
75
Co-resistance
Co-existence of multiple genes/mutations encoding resistance to different drugs within the same strain/genetic element
76
Co-selection
Selection of multiple resistance genes when one of these genes is selected
77
What is a vaccine?
A suspension of antigens that is administered to induce immunity
78
What is adjuvant?
An additive to a vaccine that enhance the immune response to the Ag
79
What are the three ways an adjuvant can enhance the immune system
1. Delay the release of Ag from the site of injection 2. Induce the secretion of chemokines by leukocytes A. Enhanced cell-mediated immunity B. Enhanced antibody production
80
What are the properties of an ideal vaccine?
``` Inexpensive Consistent in formation-minimal variations Stable Proper type of immune response Range of immunological epitope Long-lived immunity Immunological memory No adverse side-effect ```
81
What are the two types of infectious vaccines?
Live attenuated vaccines | Recombinant organism vaccine
82
Live attenuated vaccine
Attenuated, but an intact and viable organism Low-level infection No significant tissue pathology or clinical disease
83
What are the pros of a live attenuated vaccine?
Rapid onset of immunity | Sustained immunity after a single dose
84
What are the cons of a live attenuated vaccines?
Potential for reversion to virulence Virulent in the immunocompromised Less stable in storage
85
What is a recombinant organism vaccine?
Viral genes modified and inserted into a carrier | Carrier organisms do not cause disease in vaccinated animals
86
What is the benefit of a marker vaccine?
A maker vaccine permits the discrimination between a vaccine exposure and a natural immune response -in testing you can tell the difference
87
What are the non-infection vaccine types
Killed whole organism vaccine Subunit vaccine Naked DNA vaccine
88
What is a killed whole organism vaccine?
Antigenically intact | Unable to replicate or induce disease
89
What are the pros to a non-infectious vaccine?
Safe Doesnt interfere with other vaccines Stable in storage
90
What are the cons of a non-infectious vaccine?
Slow onset of immunity Multiple boosters required Adjuvant-adverse effects Reduced degree of protection vs the live
91
What is a subunit vaccine??
Contains immunological or structural proteins or metabolites of an organism Eg. Purified protein, synthetic peptides, and recombinant protein
92
What is a naked DNA vaccine?
Gene of interest is cloned into a plasmid and delivered to animal. Pathogen gene is expressed and processed in APC for antigen presentation
93
______________ immunization is performed by administering antibodies
Passive
94
Protection by passive immunization is _______________ but ______________
Immediate ; temporary
95
How do you provide active immunization?
Administer an antigen -> induce an immune response in recipient (humoral/cell-mediated response with immunological memory)
96
What are the protection levels provided with active immunization?
Strong - no infection Infected - clinically well Infected- mild form of disease Failure- no protection
97
What adverse side effects can result from vaccination?
Type I hypersensitive | Feline injection site sarcoma (FISS)
98
What are the consequences of antimicrobial resistance on animal and public health?
Increased patient mortality and morbidity | Risks of zoonotic transmission
99
What are the economic consequences of antimicrobial resistance?
More visits, lab tests, and therapies Prolonged hospitalization Reduced weight gain (food animals) Loss of customers / vet reputation Cost for hospital and farm decontamination Cost for surveillance and intervention programmes
100
What are the mechanisms of antimicrobial resistance?
Blocking of drug binding to protein target - target modification - target protection - drug trapping Change in concentration of drug in cell (mutation or membrane pores and transport systems) - increased efflux - decreased influx
101
Name three emerging resistant bacteria in animals.
``` Staphylococcus aureus (MRSA) Staphylococcus pseudintermedius (MRSP) Escherichia coli (ESBL producer) ``` Resistance to B-lactam All these bacteria are resistant to cephalosporins and are often multidrug-resistant (MDR)
102
What is MRSA?
Methicillin resistant staphylococcus aureus Resistant gene (mecA) encodes a penicillin-binding protein with low affinity to most B-lactams
103
What is MRSP?
Methicillin resistant staphylococcus pseudintermedius Resistance gene mecA 70 are skin and wound care post surgical in clinic Difficult to choose an antibiotic because of multidrug resistance Human infection usually lower but prevalence higher in dog owners and vet staff -> veterinarians as vectors for animal infection
104
What is ESBL??
Extended spectrum beta lactamase | Enzyme that can hydrolize/inactivate most b-lactams (produced by gram negative bacteria)
105
What are the three main classes of ESBL? What makes them true ESBLs
CTX-M , SHV, and TEM | Susceptible to B-lactamase inhibitors (eg clavulanic acid)
106
What is a false ESBL
resistant to B-lactamase inhibitor Widespread in Europe in poultry: CMY-2
107
How can bacteria acquire resistance?
``` Mutation (usually antimicrobial target gene/protein) Horizontal gene transfer -transformation (free DNA uptake) -transduction (phage delivery) -conjugation (cell to cell transfer) ```
108
What three antibiotics are B-lactams
Penicillins Cephalosporins Carbapenems

RUSVM Principles Of Infectious Disease (3 decks)

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