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RUSVM Principles Of Infectious Diseases > Week 8 + 9 > Flashcards

Week 8 + 9 Flashcards

1
Q

What is a vaccine?

A
  • a suspension of antigens that is administered to induce immunity
  • currently majority of vaccines derive from microbial pathogens for controlling infectious diseases
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2
Q

What does a vaccine contain?

A
  • antigens
  • preservatives + stabilizers - preserving Ag
  • specific antibiotics - inhibiting bacterial and fungal growth
  • adjuvant - enhancing the immune response to Ag
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3
Q

What is the purpose of adjuvant in vaccines?

A
  • delay release of antigen from the site of injection
  • induce secretion of chemokines by leukocytes
  • ex: aluminum hydroxide, saponin, etc.
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4
Q

What are the different types of adjuvants?

A
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5
Q

What are features of an ideal vaccine?

A
  • inexpensive
  • consistent in formation, minimal variation
  • stable
  • proper type of immune response
  • range of immunological epitopes
  • long-lived immunity
  • immunological memory
  • no adverse effects
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6
Q

What are the kinds of infectious vaccines?

A
  • live attenuated
  • recombinant organism
  • marker
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7
Q

What are features of live attenuated vaccines?

A
  • intact, viable organism
  • low-level infection
  • no clinical disease/pathology
  • pros:
    • rapid immunity
    • single dose
  • cons:
    • can become virulent
    • less stable in storage
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8
Q

What are features of recombinant organism vaccines?

A
  • use carrier organisms
  • cannot become virulent
  • adjuvant not required
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9
Q

What are features of marker vaccines?

A
  • differentiate between infected + vaccinated immune response/animals
  • ex: IBR w/ deletion
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10
Q

What are the types of non-infectious vaccines?

A
  • killed whole organism
  • subunit
  • naked dna
  • mRNA
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11
Q

What are features of killed whole organism vaccines?

A
  • antigenically intact
  • unable to replicate or induce disease
  • in chemical killing-formalin, alcohol, or alkylating agents
  • pros:
    • safe
    • stable
    • no interference w/ other vaccines
  • cons:
    • slow immunity
    • boosters
    • less protection
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12
Q

What are features of subunit vaccines?

A
  • contain immunological proteins/metabolites from a organism
    • ex: purified proteins, synthetic peptides,recombinant proteins
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13
Q

What are features of naked DNA vaccines?

A
  • gene is cloned from plasmid
  • plasmids transfer the APC’s; pathogen gene is expressed and processed in APC for antigen presentation
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14
Q

What are features of mRNA vaccines?

A
  • mRNA of gene of pathogen
  • mRNA is expressed and processed by APC for antigen presentation
  • ex: covid-19 vaccine (spike protein)
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15
Q

What are the kinds of immunization?

A
  • active + passive
  • immunization: aka vaccination, artificial induction of immunity to protect from infectious diseases
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16
Q

What are features of passive immunization?

A
  • performed (changed) antibodies administered
  • particular antigen
  • inhibit endogenous antibody response
  • sensitize recipient for hypersensitive reaction
  • immediate protection
  • fast, TEMPORARY, no memory
  • ex:
    • artificial: tetanus antitoxin, antivenoms, mAb to SARS-CoV-2
    • natural: colostrum
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17
Q

What are features of active immunization?

A
  • antigen administered
  • induced immune response (humoral/cell-mediated)
  • memory
  • protection proportional to infection severity (no infection to no protection)
  • ex:
    • artificial: vaccine
    • natural: antibody from infection
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18
Q

What are methods of vaccine delivery?

A
  • injection
  • intranasal
  • needle-free
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19
Q

What are examples of adverse effects to vaccines?

A
  • type i hypersensitivity: facial or periorbital edema, pruritus
  • FISS (feline injection site sarcoma): surgical removal challenging, poor prognosis
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20
Q

Why are diagnostic tests important?

A
  • confirm diagnosis
  • determine treatment
  • epidemiological surveillance
  • prevention, control, and eradication strategies
  • identification of new pathogens
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21
Q

What are the phases of diagnostic testing?

A
  • pre-analytical: test selection, sampling, storage, transportation
  • analytical: handling and analysis of specimen
  • post-analytical: results and interpretation
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22
Q

What is used to determine test selection?

A
  • pathogen type
  • sample type
  • test characteristics
  • disease phase
  • availability
  • cost
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23
Q

What is the difference between sensitivity and selectivity in test selection?

A
  • sensitivity: capacity of a test to correctly identify positive individuals (true positives)
  • specificity: capacity of a test to correctly identify the negative individuals (true negatives)
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24
Q

T/F: more selective tests can be used to confirm a diagnosis

A
  • false; specific
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25
Q

What are the test predictive values?

A
  • positive predictive value: probability of a positive test being a TP
  • negative predictive value: probability of a negative test being a TN
  • prevalence is very important for predictive values
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26
Q

What are different immunodiagnostic techniques?

A
  • serology: direct, indirect, titers
  • enzyme linked immunosorbant assay (ELISA): direct, indirect, sandwich
  • flow assays: lateral, bidirectional
  • agglutination
  • immunoprecipitation assays: solution based, gel based
  • immunofluorescence
  • immunohstochemistry
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27
Q

What are features of serology?

A
  • in-vitro study of antigen-antibody interactions
  • most commonly used diagnostic tool
  • basis of several techniques
  • usuall used with serum samples
  • test infectious diseases, hormones, cancer, autoimmunity
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28
Q

What are the 3 main kinds of serology?

A
  • direct: detects presence of ANTIGEN in sample, positive result = INFECTION
  • indirect: detects presence of ANTIBODIES in sample, positive result = EXPOSURE
  • titers: express concentration of antibodies/antigens, correlates with highest dilution at which they are still detectable via serial dilutions.
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29
Q
A
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30
Q

What are features of elisa?

A
  • enzyme-linked immunosorbant assay
  • uses enzyme labeled antibodies or antigens, with addition of a substrate, to detect and measure the concentration of analyte
  • direct (+ sandwich) and indirect techniques
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31
Q

What ELISA technique is this?

A
  • direct
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32
Q

What ELISA technique is this?

A
  • indirect
33
Q

What ELISA technique is this?

A
  • direct, sandwich
34
Q

What are features of flow assays?

A
  • lateral or bidirectional
  • ELISA-based
  • qualitative or semi-quantitative
  • point-of-care testing
  • fast results
  • no lab
35
Q

What are features of agglutination?

A
  • reaction between:
    • antibody + particulate antigen
    • antibody + soluble antigen coated latex beads
36
Q

What are features of immunopreciptation assays?

A
  • precipitation of antigen-antibody complexes
  • exploits the concentrations of antibody and antigen (zone of equivalence) to measure relative quantities (titers)
  • 2 methods:
    • solution based
    • gel based
37
Q

What kind of immunoprecipitation is this?

A
  • solution based
38
Q

What kind of immunoprecipitation is this?

A
  • line of precipitation on the gel indicates the zone of equivalence
39
Q

What are features of immunofluorescence?

A
  • fluorescent-labeled antibodies: tissue or cells, on slides
  • requires a fluorescent microscope
  • 2 methods: direct + indirect
40
Q

What are features of immunohistochemistry?

A
  • used on frozen or formalin-fixed tissues
  • in situ results
  • can detect tissue proteins or pathogen antigens
41
Q

What are diagnostic techniques?

A
  • direct examination
  • culture
  • immunologic methods
  • molecular analysis
42
Q

What are kinds of molecular diagnosis?

A
  • electrophoresis
  • restriction fragment length polymorphism (RFLP)
  • hybridization
  • nuclear acid amplification - target
  • protein detection
43
Q

What are features of electrophoresis?

A
  • separated in a electrophoretic field
  • negatively charged particles —> positive end
    • size: smaller is faster
    • structure: supercoiled (fastest) > linear > nicked circles (DNA)
44
Q

What are features of restriction fragment length polymorphism (RFLP?)

A
  • analyzing differences among homologous DNA sequences by restriction enzymes
  • restriction enzymes: cut DNA at the specific recognition nucleotide sequences (SPECIFIC sequence)
    - EcoRI: sticky end
    - SmaI - blunt end
  • use in paternity tests, forensics
45
Q

What are features of hybridization? Of probes?

A
  • denatured, single-stranded DNA, ssDNA, (probe) binds to a complementary single-stranded sequences
    • ex: dot, in situ, Southern, Northern, microarray
  • fragment of nucleic acids
  • labeled via radioisotope,enzyme, or chemiluminescence
  • detect complementary sequences in samples
  • high specificity
  • variable size
46
Q

What are features of nucleic acid ampification?

A
  • target amplification: enzyme mediated process to synthesize copies of targeted nucleic acid
    • polymerase chain reaction (PCR): sensitive!
      • isothermal amplification (ex: LAMP)
      • true/q PCR
47
Q

What are PCR primers?

A
  • single stranded DNA fragments complementary to sequences flanking the region to be amplified
  • distance between primer and binding sites determines size of PCR product
  • determine specificity
48
Q

What are features of primers?

A
  • types: random or specific
  • primers: product size, annealing temperature, specificity
  • nucleotide composition
49
Q

What are PCR variations?

A
  • reverse-transcriptase PCR
  • nested PCR
  • multiplex PCR
  • quantitative or real-time PCR
  • etc.
50
Q

What are features of real-time or quantitative PCR?

A
  • probe or dye to generate a fluorescent signal from the product
  • signal in real time allows quantification of starting material
  • signal: exponential curve with lag phase, log phase, and stationary phase
    • lag phase: inversely proportional to amount of starting material
51
Q

What are features of isothermal activation/LAMP?

A
  • pros:
    • no thermal cycles needed, but need room temp
    • quick - 1H
    • sensitivity > PCR
    • visible results
  • cons: complicated design of primer plates
52
Q

What are phenotypic characteristics (observable characteristics) of microorganisms?

A
  • morphology: size, shape, structure
  • biochemical reactions: staining, cell inclusions
53
Q

What are advantages and disadvantages to microscopic identification?

A
54
Q

What are features of concentration techniques?

A
  • work to increase concentration of pathogens in the sample
  • common in parasite diagnosis
  • ex:
    - flotation techniques
    - sedimentation techniques
    - baermann test for larval identification
55
Q

What are features of fecal flotation?

A
  • gross examination
  • fresh feces: detect parasites in GI tract, liver, bile ducts
  • fecal flotation: based on differences in specific gravity between the parasites eggs, larvae, cysts, or oocysts, and the majority of fecal debris
    • determine presence of parasite, egg types, general level of infection (eggs per field view)
  • McMaster egg counting slide
56
Q

What are features of the baermann technique?

A
  • method for extraction of nematode larva from fresh feces (ex: lung worm larvae in feces, larvae from fecal cultures, pastures,soil,tissues, etc.)
  • warm water stimulates live larvae in the sample to move out and gravity pulls them to the bottom of the container
57
Q

What are the two main categories of staining techniques?

A
  • simple stain (only 1 dye)
    • identification of morphology and cellular arrangement
    • drawback: may not stain all components, difficult interpretation
  • differential stain (more than 1 dye)
    • identification of morphology and cellular arrangement
    • distinguish between different cell types + structures
    • drawback: multistep method requires more time, reagents, expertise
58
Q

What are features of hematoxylin and eosin stain (H&E)?

A
  • common tissue stain used to identify a range of normal/abnormal cells + tissues, can reveal viral, bacterial, fungal, and parasitic infections
  • different structures have different dye affinities
  • hematoxylin (basic stain): stains acidic (basophilic) or - charged particles (ex: nuclei, chromatin) PURPLE
  • eosin (acidic stain): stains basic (acidophilic) or + charged components (ex: cytoplasmic components + granules) and extracellular components (ex: muscle, RBC) PINK
  • detection of VIRAL INCLUSION BODIES: aggregates made mostly of proteins, represent site of viral replication
  • virus cause virus-specific changes in cell structure, organization, and morphology
59
Q

What are features of Diff-Quick?

A
  • commercial romanowsky stain
  • used in histological rapid staining
  • fixative (fast green in methanol) > stain solution 1 (eosin G) > stain solution 2 (methylene blue)
  • air dried sample prior
  • modified wright-gives a stain: differential stain for adherence of pathogenic bacteria to cells (ex:blood cells, stain chromosomes)
60
Q

What are features of Gram-stain?

A
  • distinguish betweeen Gram + and Gram - bacteria based on CELL WALLS (peptidoglycan)
  • G+ : PURPLE
  • G - : PINK
61
Q

What are some stains for specific bacterial structures?

A
  • acid fast stain: stains organisms with impenetrable cell wall (ex: mycobacterium, cryptosporidium)
  • capsule stain: negative sting technique; contrast a translucent, darker colored background with stained cells but unstained capsule (ex: bacillus anthracis, klebsiella pneumoniae, clostridium sp.)
  • endospore stain: spores are dyed by heating malachite green dye
  • flagella stain: flagella are thickened with mordant
62
Q

What are some fungal and parasite stains?

A
  • lactophenol cotton blue stain: stain chitin in cell wall of fungi
  • gomeri methenamine silver (GMS): dark brown staining of fungal cell wall, surrounding tissue green
  • periodic acid schiff (PAS): mucin stain
  • Wheatley’s trichrome stain: detection of Protozoa
63
Q

What do cultures allow observation of? How?

A
  • mostly bacteria + fungi
  • viruses = cell cultures
  • different environmental and nutritional requirements
    • ensure pathogen growth of desired specimen
    • enrich number of pathogens for more study
    • determine pathogen identity
64
Q

What are pros and cons of culturing?

A
  • pros:
    • identification
    • assessment of AMR
    • ability of microbe to cause disease
    • study characteristic and genetics
  • cons:
    • time consuming (2-10 days)
    • expensive
    • un-culturable bacteria, no ID
65
Q

What are fastidious bacteria?

A
  • bacteria that require specific nutrients and culture conditions
66
Q

What are some kinds of fastidious bacteria?

A
  • aerobes, anaerobes, temperature dependency
67
Q

What is the great-plate culture anomaly?

A
  • ~1% of bacteria is culturally
  • unculturable: used to describe bacteria not grown on artificial media to date (not enough info)
68
Q

What are kinds of culture media?

A
  • agar (solid)
    • nutrient media: general growth
    • selective media: growth of suspected agent
    • differential media: most are selective and aid in ID
  • broth (liquid)
    • nutrient broth
    • enrichment broth: increase number of specific bacteria and limit others
69
Q

What are features of agar plate growth?

A
  • bacteria grow as colonies
  • bacterial colony: visible mass of bacteria originating from a single mother cell aka clones
  • streak plate method to obtain pure cultures
  • oxygen requirements
70
Q

What are kinds of non-selective nutrient media?

A
  • basic nutrient media
    • trypticase soy agar (TSA)
    • luria bertani (LB) agar
    • mueller-hinton (MH) agar
  • enriched nutrient media
    • blood agar
    • brain heart infusion (BHI) sugar
    • chocolate agar
    • lysed-blood agar
71
Q

What are kinds of selective nutrient media?

A
  • selective for gram-positive organisms
    • phenylethyl alcohol agar (PEA)
  • selective for fungi
    • sabouraud dextrose agar (SDA)
  • selective for gram-negative organisms
    • eosin methylene blue agar (EMB)
72
Q

What will occur if you add antibiotics to agar?

A
  • only antibiotic resistant bacteria will grow
73
Q

What are kinds of differential media?

A
  • blood agar
    • hemolysis
  • MacConkey agar
    • selective for G-
    • lactose fermentation (pink colonies)
  • mannitol salt agar
    • G+
    • mannitol fermentation (yellow colonies)
  • CLED agar: urine bacteriology
    • cysteine-lactose-electrolyte deficient
    • supports growth of common urinary pathogens (ex: proteus, klebsiella)
    • lactose fermentation
74
Q

What are biochemical tests for bacterial identification?

A
  • based on metabolic characteristics
  • enzyme production
    • catalase: breaks down H2O2
    • coagulase: causes fibrin in blood to clot
    • urease: hydrolysis urea
    • tryptophanase: ability to convert tryptophane to indole
  • carbon source utilization
  • carbohydrate fermentation: lactose, sucrose, glucose, xylose
75
Q

What are features of UTI culture paddles?

A
  • semi-quantitative culture count
  • presumptive ID of common uropathogens
  • EMB agar: selective for gram - bacteria
  • CLED agar: non-selective
  • interpretation of results
    • colony density: degree of infection
    • growth on EMB: gram -
    • color on CLED: lactose fermentation
76
Q

What are features of Flexicult Vet urinary test?

A
  • semi-quantitative colony count
  • presumptive ID of uropathogens
  • antibiotic susceptibility information
  • test within 30 mins of mid-stream urine collection, incubate at 37 C for 18-24 hours
  • > 10^3 pathogen/mL = infection
  • no growth = antibiotic susceptible
77
Q

What is the cycle of diagnostic process?

A
78
Q

What are differences between on-site analysis and diagnostic lab?

A
79
Q

What are features of a good diagnostic lab?

A

RUSVM Principles Of Infectious Diseases (6 decks)

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