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MLT Semester #1 > 1050 Finals > Flashcards

1050 Finals Flashcards

1
Q

Describe acute phase reactants

A
  • Proteins that increase due to infection, injury and trauma
  • proteins usually are:
    → c reactive protein
    → alpha-1 anti-trypsin
    → haptoglobin
    → fibrinogen
    → others, such as complement
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2
Q

What is an antigen?

A

→ foreign substance that stimulates antibody production
→ they are large complex molecules
→ molecular weight → usually more than 10,000
→ usually protein or polysacchandes in nature

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3
Q

What is an antibody?

A

immunoglobulin produced by plasma cells in response to antigen

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4
Q

What is avidity?

A

Strength of bond between antigen and antibody

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5
Q

What are chemokines?

A

→ are cytokines that attract cells to a particular site
→ important in inflammatory response

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6
Q

What is chemotaxis?

A

→ Migration of cells towards the chemokine

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7
Q

What are cytokines?

A

→ are chemical produced by activated immune cells that affect function of other cells
→ includes:
-interferons
- chemokines
-tumor necrosis factor
- transforming growth factor
- colony-stimulating factors
-interleukins

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8
Q

What is epitope?

A
  • A determinant site on an antigen
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9
Q

What are haptens?

A

-Low molecular weight
-substance that bind to antibody once its formed but is incapable of stimulating antibody’s production and less bound to large carrier molecule

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10
Q

What is histamine?

A

Vasoactive amines released from mast cells and basophils during an allergic reaction

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11
Q

What is hypersensitivity?

A

During allergic reaction hypersensitivity heightened state of immune responsiveness that causes tissue damage in the host

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12
Q

What is immunity?

A

immunity is resistance to infection

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13
Q

What is immunogen

A

Any substance capable of inducing an immune response

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14
Q

What is immunoglobulin?

A

-(Ig)
An antibody

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15
Q

What is immunology?

A

Study of reactions of hosts when exposed to foreign substance

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16
Q

What is inflammation?

A

Cellular and humoral mechanisms involved in reaction to injury or infection

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17
Q

What are interferons?

A

→ Cytokines with anti-viral properties
→ active against certain tumors and inflammatory processes

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18
Q

What are interleukins?

A

Cytokines produced by leukocytes that affect inflammatory response through increase in soluble factors i cent

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19
Q

What are ligands?

A

→ A molecule that binds to another molecule of complementary configuration
→ the substance being measured in immunoassays

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20
Q

What is Lysozyme?

A

Enzyme found in tears and saliva that attacks cell walls of microorganisms

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21
Q

What is major histocompatibility complex?

A

→ System of genes that controls expression of MHC molecules found on all nucleated cells originally
→also known as human leukocytes antigens

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22
Q

What is monoclonal? antibody

A

Antibody derived from a single b-cell clone

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23
Q

What is opsonin?

A

Serum proteins that attach to foreign substance to enhance phagocytosis

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24
Q

What is phagocytosis?

A

Engulfment of cell or particular matter by neutrophils or macrophages

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25
Q

What are plasma cells?

A
  • Transformed B cells that secrete antibody
    -in peripheral lymphoid tissues is non-dividing and they die in a few days
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26
Q

What is polyclonal antibody?

A

Antibody produced by many B cell clones

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27
Q

What is postzone?

A

→ Reduced antigen- antibody complexes due to antigen excess
→ can cause false negatives in aerological testing
For antibody
→test can be repeated in 1-2 weeks

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28
Q

What is prozone

A

→ Is a reduced antibody. Antigen complex due to antibody excess
→ this can cause false negatives in serological tests for antibody
→ dilute the semi, and retest it

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29
Q

What is seroconversion?

A

A change of serological tests from negative to positive due to the development of detectable antibodies

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30
Q

What is serum sickness?

A

Type III hypersensitily reaction that results from a build up of antibodiesto animal serum used in some passive immunizations

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31
Q

What is the thymus?

A

→ Small flat bi-lobed organ found in the thorax
→ is site of T lymphocytes development
→ one of the primary organs of lymphoid system

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32
Q

What is titer?

A

→ Means of expressing antibody concentration
→is reciprocal of the highest dilution with a positive reaction

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33
Q

What is a vaccination?

A

→ Injection of immune genetic material to induce immunity

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34
Q

What is zone of equivalence?

A

→ When number of multivalent sites of antigen and antibody are approximately equal
→ results in optimal precipitation

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35
Q

What are the branches of the immune system?

A

-humoral
- cell-mediated

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36
Q

What is humoral immune response?

A

→ Antibody mediated
→ defends against bacteria and extracellular
→ cells involved are B cells and plasma cells
→ these cells produce antibody production

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37
Q

What is cell-mediated immune response?

A

→ cell-mediated
→. Defends against viruses-fungi, mycobacteria and other intracellular pathogens and tumor cells
→ cells involved are T lymphocytes and macrophages
→ examples: graft rejection, hypersensitivity reactions and elimination of tumor cells

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38
Q

What are the steps of the humoral immune response?

A

① first exposure to antigen
② antigens engined and displayed by dendritic cells
③intact antigen activates B cells which give rise to the plasma cells and memory B cells
④these cells eventually secrete antibodies that defend against the pathogens or toxins in the extracellular fluid

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39
Q

What are the steps of cell-mediated immune response?

A

① first exposure to antigen
② antigens engulfed and displayed by dendstic cells
③ antigens displayed by infected cells and activates the helper T cells secrete the cytokines
④this activates the cytotoxic T cells and give rise to active and memory helper T cells
⑤ cytotoxic t-cells give rise to memory cytotoxic T-cells and active cytotoxic T cells
⑥this defends against infected cells, cancer cells and and transplanted tissue

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40
Q

What are the types of immunity

A

-innate (natural) defense
-adaptive (acquired) defense

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41
Q

Describe the innate defense?

A

-present at birth
- not antigen specific
- surface barriers ( skin, mucous membrane, mucus in respiratory tracts)
-internal defenses (inflammation phagocytes, NK cells fever)

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42
Q

What are the two types of adaptive defenses

A
  • humoral immunity (B cells)
  • cellular immunity (T cells)
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43
Q

What are two types of adaptive immunity?

A

-naturally acquired
-artificially acquired

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44
Q

What are the two types of naturally acquired adaptive immunity?

A

① active → antigen enters body naturally
→ body induces antibodies and specialized lymphocytes

② passive → antibodies pass from mother to fetus via placenta or to infant via breast milk

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45
Q

What are two types of artificially acquired adaptive immunity?

A

① active → antigens introduced via vaccine
→ body produces antibodies and specialized lymphocytes

② passive → preformed antibodies in immune serum are introduced by injection

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46
Q

What are the 5 leukocytes of natural immune system?

A

①basophil
②eosinophil
③neutrophil
④ monocyte/macrophage
⑤ lymphocytes

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47
Q

What the 3 granulocytes of the natural immune system?

A

①basophil
②neutrophil
⑤ eosinophil

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48
Q

Describe neutrophil

A
  • Functions for phagocytosis and inflammatory response
    -respond to chemotaxis
    -granules contain bacterial enzymes
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49
Q

Describe eosinophils

A
  • Neutralize basophil or mast cell products
    -responsible for destruction of some helmets (parasites)
    -they have some phagocytic ability
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50
Q

Describe basophils

A

-seen in hypersensitivity reactions
-they have granules that contain histamine, heparin, eosinophil chemotactic factor A
-in allergic response, they bind to IgE and granules release contents in the presence of the antigen

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51
Q

Describe monocytes

A

-responsible for phagocytosis
-they migrate to tissues and become macrophages
-respond to chemotaxins

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52
Q

Describe mast cell

A

-A tissue cell
-responsible for hypersensitivity reaction
-seen in connective tissue cells
-resemble basophils but are larger and they contain more granules and bind to IgG

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53
Q

Describe macrophages

A

-phagocytosis
-help eliminate bacteria, intracellular parasites, tumor cells
- secretion of cell mediators
- antigen presentation
-activated by contact with microorganisms or cytokines from T lymphocytes

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54
Q

Describe dendritic cells

A

-phagocytosis in nature
-responsible for presentation of antigen to helper T cells in the blood and lymph organs
-initiate acquired immune response.
- tissue cells with long membranous extensions

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55
Q

Describe Natural killer cells (NK)

A
  • First line of defense against tumor cells and cells infected with viruses
    -lymph’s without T or B markers
    -CD16 and CD56 positive on cell surface
  • bridge between acquired and innate immunity
  • lack specificity
  • stimulated by cytolones
    -respond early infection which allows time for the T and B cells to be activated
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56
Q

What are the cells of the acquired immune system

A

-T lymphocytes
- plasma cell
- killer cells

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57
Q

Describe T lymphocytes

A
  • function of cell mediated immunity
    -derived from cells in bone marrow
    -they develop T cells specific surface antigen in the same thymus
    -60 - 80 % lymphocytes are T lymphocytes
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58
Q

Describe helper inducer cells

A
  • Orchestrate cell mediated immunity,
  • activate B cells, cytoxic cells and NK cells
    -CD4 positive
  • 2/3 peripheral T cells
  • normal CD4 is about 1,000 per milliliter, in AIDS → > 200 per milliliter
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59
Q

What is attenuation?

A

→ A process of producing nonpathological bacteria or rinses for use in vaccines
→ organisms have been weakened by treatment with chemicals, changed temperature or repeated passage in cell culture

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60
Q

What are chemotaxins?

A
  • A protein or other substances that acts as a chemical messenger to produce chemotaxis
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61
Q

What is diapedesis

A

Process by which cells are capable of moving from the circulating blood to the tissues by squeezing through the wall of blood vessel

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62
Q

Describe germinal center

A

Interior of secondary follicle where blast transformation ot B cells take place.

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63
Q

Describe memory cells

A
  • Progeny of an antigen activated B or T cells that is able to respond to antigen more quickly than the parent cell
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64
Q

Describe primary follicle

A

A cluster of B cells that have not yet been stimulated by antigen

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65
Q

Describe primary lymphoid organs

A

-the organs in which lymphocytes mature: these are bone marrow and the thymus

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66
Q

Describe secondary follicle

A

→ A cluster of cells that are proliferating in response to specific antigen.
→ has germinal center and stimulated B cells

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67
Q

Describe secondary lymphoid organs

A
  • Where the main contact with forgein antigens takes place
  • spleen, lymph nodes, appendix, tonsils and other mucosal-associated lymphoid tissue
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68
Q

Describe spleen

A
  • Largest secondary lymphoid organ
  • filters out aged cells and foreign
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69
Q

Describe thymocytes

A

-immature lymphocytes
-found in the thymus
-undergoes differentiation of T cells to become a mature cells

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70
Q

Describe complement

A
  • Series of proteins (30 or more) that are normally present in serum
    -overall functions are medication, osponization, chemotaxis, cell lysis or inflammation and destruction of foreign cells
    -most are inactive enzyme precursor that ave converted to active enzymes in precise order
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71
Q

Describe defenses

A
  • Small canonic proteins
  • released by lysosomal granules
  • can kill bacteria and many fungi by destroying their cell walls
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72
Q

Describe fibrinogen

A

-acute-phase reactant that changes to fibrin
-forms clot in the bloodsteam

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73
Q

Describe haptoglobin

A
  • An acute-phase reactant
  • binds irreversibly to free hemoglobin released by intravascular hemolysis.
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74
Q

Describe opsonins

A
  • Serum proteins that attach to a foreign substance and enhance phagocytosis
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75
Q

Describe oxidative burst

A
  • Increase in oxygen consumption in phagocyticceas which generate oxygen radicals used to kill/engulf microorganism
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76
Q

Describe Pathogen-associated molecular patterns (pamps)

A

-.structural patterns of carbohydrates, nucleic acids, bacterial peptides or microorganism
- recognized by the pathogen recognition receptors on the cells of the innate immune system

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77
Q

Describe toll-like receptors

A

-receptors found on human leukocytes and other cell types
-recognize microorganisms and aid in their digestion

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78
Q

Describe adjuvant

A

Substance administered with a immunogen that enhances and potentates the immune system

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79
Q

Describe alloantigen

A
  • Antigen that is found in another membrane of the hosts species
  • capable of eliciting an immune component host
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80
Q

Describe autoantigen

A

Antigen that belongs to the host and is not capable of eliciting an immune response under normal control circumstances

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81
Q

Describe hapten

A
  • A small molecule which, when combined with a larger carrier such as a protein, can elicit the production of antibodies which bind specifically to
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82
Q

Describe heteroantigens

A

An antigen of a species different from that of the host, such as other animals,, plants, or microorganism

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83
Q

Describe antigen dependent stage

A

-final phase of B cell development in bone marrow that results in mature B cells that have not been exposed to antigen.

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84
Q

Describe antigen- independent phase

A

-If a B call is stimulated by antigens it undergoes transformation to blast stage that eventually forms memory cells and antibody secreting plasma cells

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85
Q

Describe heterophil antigen

A

Antigen that exists in unrelated plants or animals but either identical or closely related so that antibody to one Will cross- react with antibody to the other.

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86
Q

Describe suppressor T cells

A
  • Inhibit helper T cells and cytotoxic cells that kill other cells
  • CD8 positive
  • 1/3 of peripheral T cells
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87
Q

What is the normal ratio of CD4 and CD8? What is it in AIDS?

A

Normal → 2:1
AIDS → 0.5:1

88
Q

Describe T regulatory cells

A
  • Suppress immune response to self
    -CD4 and CD25 positive
89
Q

Describe B lymphocytes

A
  • After antigenic challenge → transform into blast cells that give rise to plasma cells and memory cells
  • develop in bone marrow
    -when mature, they have surface IgM or IgD which act as receptors for antigens
  • 10 - 20% of lymphocytes ave B cells
90
Q

Describe memory B cells

A
  • Respond to antigen when encountered again with an increased speed and intensity
    -in peripheral lymphode organs they live months to years
91
Q

Describe the antibody structure

A

-two heavy chains and light chains hold together by disulfide bonds
- bond determines class
-FAB fragment ( each have 2)
-FC fragment

92
Q

What are 2 types of light chains in an antibody structure?

A
  • Kappa
    -lambda
  • found in all classes but only one type per molecule
93
Q

free L light chains is also called what?

A
  • Bence jones proteins
94
Q

Describe FAB fragment

A
  • Fragment antigen binding it consists of one light and one heavy chain held together by the SS bonds
    -a per antibody, each can bind the antigen
95
Q

Describe FC fragment

A

-crystallizable terminal
- carbonyl (spelling?)terminal carboxy halves ot two heavy chains held held together by Disulfide bonds
-plays a role in ospinization and complement fixation

96
Q

Describe constant region of an antibody structure

A

-carboxyl terminal ends of the heavy and light chains where nano acid sequence is the same for all chains of that type

97
Q

Describe variable region of antibody structure

A

-amino terminal ends of the heavy and light chains where anime and sequence varies
- aka antigen recognition unit
-responsible for antibodies specificities

98
Q

Describe hinge region of antibody structure

A

-Flexible portion of heavy chain between first and second constant region
-allows molecules to bend so that two antigen binding site can operate independently

99
Q

Describe Joining chain of antibody structure

A

-glycoprotein that links the antibody monomers to IgM and secretory IgA

100
Q

Describe IgG

A
  • Monomer
    -150,000 moleuker weight
    -heavy chain → gamma
  • light chain → kappa or lambda
    -70-75% of immunoglobulins
    -serum concentration is 800- 1600 mg/dl
  • 2 antigen binding sites
    -will fix complement
    -will cross placenta
  • role→ defense against bacteria/viruses,neutralizes toxins, opsionin antigens, and part of passive immunity in new born
    -more effective at precipitation than agglutination
101
Q

Describe IgM

A

-pentamer → largest
- 900,000 molecular weight
- heavy chain is mu
- light chain → lambda or kappa
- 10% of total antibody
-serum concentration → 120-150 mg/dl
- 10 binding sites
- fixes complement
-neutralize toxins and opsonin
-first produce in bodies immune response
- only one produced by newborn
-most efficient at initiating complement cascade
-move efficient in agglutination than IgG
- destroyed by sulfa-dial compounds

102
Q

Describe IgA

A
  • Serum structure → monomer
  • Secretions → dimer
  • Monomers → 160,000 molecular weight
    -Dimer → 400,000 molecular arweight
  • heavy chain → alpha
  • light chain → lamba or kappa
  • serum concentration→ 70-350 mg/dl
    -10-15% total antibody count
    -either 2 or 4 antigen binding sites
  • does not fix complement
    -first line of defense,patrols mucosal surfaces, prevents adherence of bacteria, and neutralizes toxins
  • can be found in tears, sweat, saliva, breast milk, and respiratory and GI mucousa
103
Q

Describe IgD

A

-monomer
-180,000 molecular weight
- heavy chain is delta
-light chain is lambda or kappa
-serum concentration → 1-3 mg/di
- >1% total antibody count
- 2 antigen binding sites
- does not fix complement
-B cell maturation
-found on surface of B lymphocytes

104
Q

Describe IgE

A

-monomer
- 190,000 molecular weight
- heavy chain is epsilon
- light chain is kappa or lambda
- 0.002 % total antibody count
-serum concentration→ 0.005 mg/dl
-2 antigen binding sites
- does not fix complement
- role is in allergic reaction→ can bind to the basophil or mast cells
- when two adjacent molecules on mast cans bind the antigen → degranulation of the cell with release of histamine and heparin
-found in type I immediate hypersensitivity reactions

105
Q

Describe classical pathway

A

-trigged by an antigen-antibody reaction
-IgM is most efficient activator
- single molecule attached to a adjacent antigens can initiate the cascade
- IgG1, IgG2, and IgG3 can activate complement but at least 2 molecules are required
- recognition unit →C1
- activation unit →C4, C2, C3
- membrane attack complex→ C5, C6, C7, C8, and C9 causes cell lysis

106
Q

Describe alternative pathway

A
  • Antibody independent
  • activated by bacteria, fungi, viruses, tumor cells, and some parasites
107
Q

Describe lectin pathway

A
  • Antibody independent the
    -initiated by mannose binding lectin (MBL)
  • nonspecific recognition of sugars on microorganisms
    -important defense mechanism during infancy
  • C3 in highest concentration in our plasma
108
Q

What is the key component of all 3 complement pathways?

109
Q

What ions are required for the cascade of complement?

A

Calcium and magnesium

110
Q

What do deficiencies in complement pathway cause?

A
  • Increased susceptibility to infection
  • accumulation of immune complexes which can lead to glomenuar nephritis
111
Q

What are the agglutination methods?

A

① direct agglutination
② hemagglutination
③ passive (indirect) agglutination
④ reverse agglutination
⑤agglutination inhibition
⑥ hemaggietination inhibition
⑦ coagulation
⑧antiglobusin - mediated agglutination

112
Q

Describe direct agglutination

A
  • Naturally occurring antigens on particles
    -particles agglutinate in the presence of a corresponding antibody
    -examples
    → widal test for typhoid fever, salmonella
    →O and H antigens used to detect antibodies in patient scheme
113
Q

Describe hemagglutination

A

-this is antigen and antibody reaction that results in clumping of RBCs
-examples
→ ABO slide typing

114
Q

Describe passive (indirect) agglutination

A
  • Soluble antigens bound particles
  • example
    → latex and particles of gluten and in the presence of a corresponding antibody
    →rheumatoid factor
    → anti-nuclear antibody
115
Q

Describe reverse passive agglutination

A
  • Antibodies attach to carrier particles
    -particles agglutinate in presence of the Corresponding antigen
    -rapid ID of bacteria is used with this type of testing
116
Q

Describe Agglutination inhibition

A
  • competition between particulate antigens such as a reagent and soluble antigen into specimen for sites on a reagent -antibody
    -lack of agglutination is a positive result in this test
  • can be used in detection ofinicitargs
117
Q

Describe hemagglutination inhibition

A
  • Detects antibodies to certain viruses that agglutinate RBCs
  • in presences of antibody, virus is neutralized and hemagglutination does not occur
    -examples
    → rubella and other viruses
118
Q

Describe coaggulation

A
  • Reagent antibody is attached to carrier bacteria
  • staph aureus most frequently used carrier
    -protein A binds the fc portion of the reagent antibody
    -visible agglutination in presence of corresponding antigen
    -example
    → rapid ID of bacteria
119
Q

Antiglobulin-mediated agglutination

A
  • Detection of non-agglutinating antibodies by coupling with second antibody anti-human globulin
  • example
    → direct and indirect anti-globulin test
120
Q

What are the precipitation methods’

A

①ouchterlony double diffusion
② radial immunodiffusion (RID)
③ Rocket immunoelectrophoresis
④ immunoelectrophoresis (IEP)
⑤ immunofixation electrophoresis (IFE)
⑥ nephlometry

121
Q

What is precipitation?

A
  • Soluble antigen that combines with soluble antibody to produce visible complex
  • less sensitive than agglutination
122
Q

Describe ouchterlony double diffusion

A
  • Antigens and antibodies diffuse from wells in gel and form precipitation lines where they meet
    -examples
    → fungal antigens
    → extractable nuclear antigens
123
Q

Describe radial immunodiffusion

A
  • Antigen diffuses out of the well in gel containing the antibody
    -precipitation ring forms
  • diameter is proportional to the concentration of the antigen
  • examples
    Flow volume testing for IgD and lgG
124
Q

Describe rocket immunoelectrophoresis

A

-electrical charge applied to RID to facilitate migration of antigen into the auger
- height of the rocket shaped precipitation band is proportional to the concentration of the antigen
- examples
→ antigen IGs or antibodies complement
→ alpha-fetoprotein

125
Q

Describe immunoelectrophoresis

A
  • Proteins separated by electrophoresis than double diffusion with reagent antibodies in trough in the auger
  • shape, intensity, and location of the precipitation arcs are compared with a normal control
  • example
    → serum proteins including antibodies
126
Q

Describe immunofixation electrophoresis

A
  • Proteins separated by electrophoresis
  • anti-serum is placed directly on the gel and antibody complexes precipitate out
  • examples
    →IDs of antibodies and monoclonal gamonopathies
    → pence jones proteins and western blot
127
Q

Describe nephelometry

A
  • Light is scattered by the antigen antibody complexes
  • used to detect antibodies complement and o reactive protein
128
Q

Describe isotopic immunoassay

A

-immunoassay that uses radioisotopes as the label

129
Q

Describe non-isotopic immunoassay

A

-uses something other than radioisotope as a label
- could be enemies, florachrome, chemiliminescent molecules

130
Q

Describe competitive immunoassay

A

-Patent antigen and a labeled reagent antigen compete for binding sites on reagent antibody

131
Q

Describe non-competative immunoassay

A
  • Does not involve competition for binding sites
  • more sensitive than competitive assay
132
Q

Describe heterogenous immunoassay

A
  • has separation step to remove the free form bound analyte
  • more sensitive than homogeneous
133
Q

Describe homogenous immunoassay

A
  • That doesn’t require separation state
  • easier to automate
134
Q

What are the 3 types of enzyme immunoassays?

A

①direct EIA
②indirect EIA
③solid phase

135
Q

Describe EIA

A

-Uses enzymes as the label
- substrate is added to measure enzyme activity

136
Q

Describe direct EIA

A
  • Competitive enemy labeled reagent is part of initial antigen-antibody reaction
    -all reactants are added at the same time
    -incubation and then Wash step
137
Q

Describe the indirect EIA

A
  • Non competitive
  • enzyme labeled reagent isn’t involved in the initial antigen- antibody reaction
    -two incubations and two washes
    -more sensible than ELISA
138
Q

Describe solid phase

A

Reagent antigen or antibody that 1s bound to support medium, could be polystyrene testtubes, microtiter plates, cellulose membranes and glass beads

139
Q

What are the formats of EIA?

A
  • Heterogenous
    -competitive
    -direct
    -enemy labeled ligand and unlabeled patient ligand compete for binding sites on antiboy attached to a solid phase
    -free labeled ligand is removed by washing
  • substrate is added, color is inversely proportional to the concentration of the ligand and its specimen
  • used to measure small relatively pure antigens for
    → examples: insulin and estrogen
140
Q

What are the formats of ELISA?

A
  • Heterogenous
    -non competitive
  • indirect
  • antigen is attached to solid phase
  • antibody and specimen attaches
  • unbound antibodies are removed by washing
    -then enzyme labeled anti-globulin is added
    -attaches to antibody through solid phase
    -substrate is added, color is directly proportional to antibody concentration
    -move sensitive than competitive EIA
  • most common immunoassays
  • used to detect antibodies to viruses such as HIV, hepatitis A, C, and epstein-barr virus
141
Q

Describe sandwich ELISA/ capture assay

A
  • Heterogenous
    -noncompetitive
  • indirect
  • antibody attaches to solid phase
    -antigen in specimen attaches
    -enzyme labeled antibody is added which attaches to different determinant
    -Enzymatic activity is directly proportional to the amount of antigen in sample
  • antigens must have multiple determinants
  • used to measure antibodies, hormones,proteins, tumor marker, viruses, parasites,
    and fungi
    -high concentration of antigen can cause that hook effect → undiluted samples have lower absorbence
142
Q

Describe rapid ELISA

A
  • Membrane-based ‘
  • reagent antigen or antibody is bound to the membrane in single use cassette
    -sample is added the presence of antigen- antibody complex is indicated by a colored reaction
    -built in quality control
    -usually qualitative
143
Q

Describe EMIT format

A
  • Homogenous
  • antigen in specimen and enzyme labeled antigen compete for binding sites on a reagent antibody
  • when enzyme labeled antigen binds → enzyme activity is inhibited
    -enzyme activity is dircetly proportional to concentration of the antigen in specimen
    .used for determinations of low molecular weight analytes not readily measured by other methods ( hormones, therapeutic drugs, abused drugs)
  • also can be used in automated format
144
Q

What are the 3 fluorescent immunoassays

A

① direct (DFA)
② indirect (IFA)
③ polarization (FPIA)

145
Q

Describe direct fluorescent immunoassay

A
  • Antibody staining
  • specimen on glass slide overlaid with a fluorescent labeled tag or antibody.
    -is corresponding antigen is present, the labels antibody will bird.
    -then fluorescence is observed with a fluorescent microscope
  • can detect antigens fluorescents labels can be fluorescein, isothiocyanate and rhodamine B isothiocyanate
  • can also be used for bacterial and viral antigens
146
Q

Describe indirect Fluorescent immunoassay

A

-antibody staining
- reagent antigen in the serum
on glass slide overlaid with patient serum
- if corresponding antibody is present in the serum
It attaches to the antigen
-when fluorescine labeled anti-human globulin is added, it will attach to the antibody
-fluorescence is observed again with your fluorescent microscope→ this is called a sandwich technique
-this technique and it will detect the antibodies to the particular virus
-examples
→used for fluorescent anti-nuclear antibody fana(sp?)
→fluorescent treponemal antibody (FTA)

147
Q

Describe fluorescent polarization immunoassay

A
  • Labeled antigen that will compete with an antigen in specimen for sites on reagent antibodies
    -free labeled antigen rotates rapidly and admits little polarized light
    -the bound labeled antigen rotates more slowly and will admit more polarized light
  • amount of polarized right is directly proportional to the concentration of the antigen in the speciemen
  • can be used as competitive homogenous and can be automated
  • used to detect therapeutic drugs and hormones
148
Q

What are the labeled immunoassay?

A

①RIA
②EIA
③FIA
④CIA

149
Q

Comparison of RIA labeled immunoassay

A
  • Labels arc 125 and 131 iodine detection
  • can defect radio isotopes that admit radioactivity
  • the types are competitive and heterogenous
  • advantages → its sensitive and specific
  • disadvantages → radiation hazard, short shelf life of reagents, disposal problems and licensing and federal regulations
  • not commonly used anymore
150
Q

Companion of EIA labeled immunoassay

A
  • Label can be alkaline phosphates, horseradish peroxides, diactylase glucose 6-phosphate dehydrogenease
  • used to detect enzymes that react with substrate to produce color change
    -mostly non competitive
  • heterogenous and homogenous are available
    Advantages→ sensitive, Specificity, no health hazard
    Or disposal problems, reagents with long shelf life and it can be automated
    Disadvantages → natural inhibitors in some specimens nonspecific protein binding
151
Q

Comparison of FIA labeled immunoassay

A

-labels are fluorisim and rhodammine
-used to detect Floor chromes that absorb energy from light source and can convert to longer wavelengths or lower energy
- assays available are usually competitive
- can be both heterogeneous and homogenous
-advantages → automated, sensitivity, specificity, long reagent shelf life
-disadvantages → autofluorescence from organic substances in serum, nonspecific binding to substances in the self, expensive and have dedicated instrument
- commonly used

152
Q

Comparison of CIA labeled immunoassay

A

-labels can be luminal, acronym, esters, and nitrophenyloxalate
-used to detect chemiluminescent molecules that produce light from chemical reactions
- competitive and non competitive available
-heterogenous and homogenous
- advantages→ sensitive, specificity, automated, long reagent shelf life
.disadvantages → quenching of lightemission by some biological materials
- commonly used

153
Q

Describe hypersensitivity type I

A

-IgE- mediated
-“anaphylactic”
- release of mediators from most cells and basophils
- on set of symptoms is immediate
-examples
→ anaphylaxis
→ hay fever
→ asthma
→ food allergy

154
Q

Describe hypersensitivity type II

A

-“cytotoxic”
- key reactants are IgG or IgM complement and cellular antigens
- cyto lysis due to antibody and complement
- immediate of onset symptoms
- examples
→transfusion reactions
→ hemolytic disease of a newborn
→ autoimmune hemolytic anemia

155
Q

Describe hypersensitivity type III

A

-is immune complex
- it includes IgG, IgM, complement and soluble antigens
- it deposits antigen antibody complexes in the tissues
-immediate onset of symptoms
- examples
→ artherus reaction
→ serum sickness
→SLE
→ rheumatoid arthritis

156
Q

Describe hypersensitivity type IV

A

-T cell dependent
-key reactants → T-cells
- release of cytokines
- onset of symptoms is delayed
- sensitization after first contact with antigen
And then symptoms upon re-exposure
- examples
→ contact dermatitis hypersensitivity
→ pneumonitis
→ tuberculous skin test

157
Q

What are 2 nontreponemal tests for syphilis

A

①VDRL
②RPR

158
Q

Describe VDRL (syphilis)

A

-flocculation→ it detects reagin which is the antibodies against cardiolipin that is in serum of patients with syphilis.
- antigen is cardiolipin
- positive reaction is microscopic clumps
-specimen is inactivated serum or spinal fluid
- reactivity during disease may be negative in primary stage
-titer usually peak during secondary and early late stages
-titer in late stages,even untreated, more rapid decline with treatment
-becomes non reactive one to two years following
Successful treatment now
-false positives→ can be biological with infectious mono, infectious hepatitis, malaria, leprosy, lupus, rheumatoid arthritis, advanced age and pregnancies
- screening test and should be confirmed treponema test

159
Q

Describe RPR (syphilis)

A
  • Flocculation → detects reagin
    -antigen is cardio lipin combined with charcoal
    -macroscopic agglutinate
  • speciemen are serum
    -inactivation not required
  • same reactivity and false positives as VDRL
  • used for screening and treatment monitoring
  • for screening test → reactive should be confirmed by treponema test
160
Q

What are the 3 treponomal test for syphilis?

A

①FTA-ABS
② TP-PA
③ELISA

161
Q

Describe FTA-ABS for syphilis

A
  • Antibody absorbent test
  • defects antibody to T. Palladium
    -reagents
    →absorbent non-pathogenic treponema such as reiter stain
    → slides with Nichols’ strain of t, palladium
    → fluorescences labeled anti-human globulin
    -a positive reaction in this is the fluorescence
  • specimens→ serum and spinal fluid
  • reactivity of disease is usually positive before Reagan tests are.
    -false negatives→ primary syphiliscusually positive for life)
162
Q

Describe TP-PA (syphilis)

A
  • Antibody is to t, palladium
    -reagents → colored gelatin particles coated with treponemal antigen
    -positive reaction is an agglutination of sensitized gel particles.there’s a smooth over a surface of wells
    -specimen is serum
  • reactivity→ not as sensitive in primary syphilis as FTA, sensitivity y is close to 100% in secondary syphilis and positive in late stages
163
Q

Describe ELISA (syphilis)

A

-Detects antibody to T palladium
- reagent is enzyme labeled treponemal antigen
-positive reaction is color development following addition of substrate
- speciemen is serum
- reactivity→ highly sensitive in primary syphilis but decreases in later stages

164
Q

If RPR and FTA are reactive, what is the interpretation?

A
  • positive for syphilis
165
Q

If RPR is reactive and FTA is non-reactive, what is the interpretation

A
  • Negative for syphilis
166
Q

Describe serological testing for anti-streptolysin O (ASO)

A

-group A strep infection, rheumatic fever, and post streptococcal glomerular nephritis
- common test is nephlometry
- uses recombinant streptomycin antigen
- if antibody is present, antigen-antibody complex that forms and an increase in light scatter
-replaces classic neutralization method

167
Q

Describe serological test for Anti-DNase B

A
  • Diagnoses sequel of group a strep, rheumatic fever, glomerular nephritis following skin infection
  • common methods→ EIA and nephlometry
  • High specific
    -may be positive when ASO is negative
168
Q

Describe serological test of streptozyme

A
  • Diagnosis sequel of group a strep infections
  • slide agglutination test it
    -uses RBCs coated with several streptococcal antigens
    -moRe false positives and false negatives Are seen with this test
  • should be used with ASO and antii-dnase B
    -serial titers should be performed with this
169
Q

Describe serological testing for helicobacter pylori antibodies

A
  • Diagnose gastric and duodenal ulcers caused by H. Pylori
  • method of testing → ELISA, rapid test, and PCR
  • most tests detect IgG
    -25% decrease in titer→ successful treatment
  • antibodies will remain for a year
    -positive rapid test should be confirmed by ELISA
170
Q

Describe serological tests for mycoplasma pneumonia

A
  • Diagnosis primary atypical pneumonia
  • most common test methods → EIA, agglutination, IFA, and molecular methodsthat are available
  • contest for IgM and IgG antibodies
  • replaces cold agglutinin ( was non specific)
171
Q

Describe serological test for rickettsial antibodies

A
  • Diagnose typhus, Rocky Mountain spotted fever and others
    -gold standard is IFA and micro-IF
    -PCR is available
172
Q

What are the 2 tests for infectious mono

A

① heterophil antibodies
②EBV - specific antibodies

173
Q

Describe serological testing for heterophile antibodies (IM

A
  • Nonspecific antibodies that agglutinate horse, sheep and bovine RBCs
  • heterophile antibodies→ antibodies that react with similar antigens from different species
  • occurrence → 90% of patients develop first n the first month of illness
  • can persist for one year
    -negative and 10% ot adults and up to 50 percent of children with IM
    -if they are symptomatic and heterophile negative→ test for EBV- specific antibodies
    -testing method→ rapid latex agglutination, solid phase immunoassays
  • antigen is a puntied bovine RBC extract and typically screening tests
174
Q

Describe EBV- specific antibodies

A

-this has specific antibodies against epstein-barr virus
-these antigens present in different phases of infection
-early → early antigen
- late → is viral capsule capsid antigen which is VCA
-Latent→EBV nuclear antigen (EB-NA)- occurance - the anti -VCA IgM appears at onset of symptoms and disappears in about 3 months
- anti-VCA IgG appears at onset of symptoms and will persist for life
-EB-NA present during convalensenCE
- acute infection would include the anti-VCA IgM and IgG and anti-EA.
- past infection →anti-EB-NA, anti -VCA IgG
-testing method → indirect immunoflurence assay, ELISA chemo luminouscent assay
-IFA is gold standard but time consuming and harder to interpret
- molecular testing use for immunocompromised patients

175
Q

Describe hepatitis A and test

A
  • Test for the total anti-hepatitis A virus
  • indicates past infection and immunity
  • can test for IgM anti-HAV→ acute infection
176
Q

Describe hepatitis B tests

A
  • Can test hepatitis B surface antigen → signify acute or chronic infection or infectivity
    -first serological marker to appear
  • used to screen donor blood
  • positives should be confirmed with repeat testing and another assay such as hepatitis B surface antigen neutralization or hep B virus DNA
  • hepatitis BE antigen→ signifies acute or chronic infection and indicates high degree of infectivity
  • total anti-HBC → signifies current or past infections or corner.
  • predominantly IgG and will persist for life
  • IgM anti- hb core → current or recent infection, first antibody to appear
  • detect hbv infection when the hepatitis B surface antigen is no longer detected (window period)
  • anti-hbe→ signifies recovery and reduced infectivity
    -anti-HBS→ signifies recovery and immunity
    -hep B test → DNA→ signifies current infection, detected 21 days before the HBS antigen and use to monitor viral load
177
Q

Describe hepatitis C test

A
  • Anti -HCV → signifies acute, chronic, and previous ‘ infection
  • positive should be confirmed by recombinant immunoblot assay
    -HCV RNA → indicates current infection
    -Used for new 1009 testing, blood organ donor screening,
    -HCV genotypingto determine optimal treatment
178
Q

Describe hepatitis D tests

A

-IgM anti-HDV → acute or chronic infection
-HDV is a defective viRUS that can only outer in presence of hep B
-IgG anti-HDV → indicates recovery or chronic infection
-HDV RNA → current infection
- marker of active vi therapy ral replication
- used to monitor

179
Q

Describe hepatitis E tests

A
  • Currently not approved by FDA for use in the United States
180
Q

Hepatitis A profile

A

Acute → IgM anti-HAV+
Recovery→ total anti-HAV+

181
Q

Describe profile for acute hepatitis B

A

-HBS antigen +
-total- anti-HBC +
-IgM anti HBC +
-anti-HBS -

182
Q

Describe profile for recovery from hep B

A

-HBS-antigen+
-total anti-HBc+
-anti-HBs+

183
Q

Describe profile chronic hepatitis B/ carrier

A

-HBs- antigen+
-total anti-HBc+
-IgM anti-HBc-
-Anti-HBs-

184
Q

Describe profile of Hep B immunization

A
  • HBs- antigen-
    -anti-HBc-
    -anti-HBs+
185
Q

What are HIV markers?

A
  • Viral RNA → is detectable within days of infection
    -PG-24 antigen→ cove code for nucleic acids, detectable in 2 to 3 weeks and becomes undetectable as antibodies are developed and detectable again in late stages
  • IgM antibody → usually detected in 2-8 needs and its transient, peaks at 1-2 weeks and undetectable 1-2 weeks later
    -lg antibody → detectable shorty after the IgM antibody, gradually increases n titer for several months and its also long lasting
186
Q

What are HIV screening test?

A

①EIA/ELISA
②rapid tests
③ nucleiC acid amplification testing (NAAT)

187
Q

Describe EIA screening for HIV

A

-first generation →IgG antibodies to HIV one only last 6-12 weeks)
-second generation → lgG antibody to 1 and 2 types of HIV
- third generation → IgG and IgM antibodies to HIV 1 and 2. Detected in 3-4 weeks
-fourth generation →(current) IgG and IgM antibodies to HIV 1 and 2 plus testing for p24 antigen (detect in 2 weeks)

188
Q

EIA/CIA p 24 antigen without HIV antibody indicates what?

A

An acute infection

189
Q

EIA/CIA p 24 antigen with HIV antibody indicates what?

A

Established infection

190
Q

Describe rapid tests for HIV screening

A
  • Includes IgG and IgM antibody to HIV which can pick up w/ an infection 4-12 weeks
    -testing → immunochromatic graphic assays
  • Specimen → blood, serum, and oral fluids
191
Q

Describe NAAT HIV screening

A

-HIV RNA→ window period is 5 days
-not cost effective
- used for screening blood donors and infants

192
Q

What are false positives in ELISA testing for HIV?

A
  • Heat inactivation of serum
  • repeated freezing /thawing of serum
    -autoantibodies
    -multiple pregnancies
  • liver disease
    -administration of immunoglobulin
  • administration of certain vaccines
  • some malignancies
193
Q

What are false negatives in screening for HIV?

A
  • Blood drawn before seroconversion
  • hypogammaglobulinemia
  • immunosuppressive therapy
  • strain of HIV not detected by assay
  • technical errors
194
Q

What are the HIV confirmatory/supplemental test?

A

① western blot
②IFA
③NAAT

195
Q

Describe western blot HIV test

A
  • detects antibodies to HIV
    -traditional confirmatory test but not as sensitive as EIA anD NAAT
    -interpretation is controversial because most labs report positive it atleast 2 of the following 3 bands ave present →P24, gp41, gp120 or gp160
    -NAAT required folloWING negative or indeterminant results
  • time consuming and difficult to interpret
196
Q

Describe IFA for HIV screening

A
  • detects antibody to HIV
  • sensitive and specific compared to western blot
    -expensive and subjective in testing results
197
Q

Describe NAAT testing for HIV

A
  • Detects HIV RNA specifically
    -qualitative test used for conformation
198
Q

What are the tests used to stage and monitor HIV?

A

① CD4 T cell count
②HIV-1 viral load assays

199
Q

Explain CD4 T cell count for monitoring HIV

A

-HIV infects CD4 cells, number declines as disease ‘ progresses
- >200 per microliter defines AIDS
-used to monitor therapy to see iF cells were declining or staying steady or improving
-performed every 3-6 months
-flow cytometry is gold standard

200
Q

Describe HIV-1 viral load assays

A
  • Qualitativ eNAAT to determine pkesma HIV RNA
  • used to predict progression, determine when to start retro-viral therapy and monitor response to therapy
    -test 2-8 weeks after start of therapyand then every 3-4 months
201
Q

What are the screen in test for SLE?

A

① fluorescent anti-nuclear antibody (FANA)
②EIA anti-nuclear antibody (EIA -ANA)

202
Q

Describe fluorescent antinuclear antibody (FANA) screening for SLE

A

-high sensitivity → 95-100%
- low specificity
- antibodies present in other autoimmune diseases can interfere or cause false positive
-2% of healthy individuals and 75% of the elderly will also test positive
-dilution tested to eliminate lower titer reactions in normal population
-cut off for dilution is greater than or equal to 180
-end titer should be reported and are usually higher in SLE patents.

203
Q

Describe indirect immune fluorescence of SLE screening

A
  • Substrate of human epithelial tumor on line hep-2
  • detects antibody to greater than 100 autoantigens
    -Staining patterns do not need to be reported anymore
    -very labor intensive and subjective
    -still considered the gold standard
204
Q

Describe EIA antinuclear antibody testing for SLE

A
  • Not as sensitive
  • specificity is low
    -Easier to perform and less expensive
  • can be automated
    -interpretation is not subjective but notas many antigens we can test for
205
Q

What are tests for specific antinuclear antibodies?

A

①anti-dsDNA
②anti-sm
③antihistone, anti-DNP, anti-ss-ARo, anti-ss-B/la and anti-nRNP
④extractable nuclear antigen antibiotics (ENA)

206
Q

Describe anti-dsDNA testing for specific anti-nuclear antibodies

A
  • Low sensitivity but high specificity for SLE because its uncommon in other diseases or normal individuals
  • most specific antibody for SLE
    -titers correlate with disease activity
  • peripheral of fluorescent pattern is seen in IFA
  • other methods → EIA, immunoblotting, and immune diffusion
207
Q

Describe anti-sm test for specific anti-nuclear antibodies

A

-has low sensitivity, but high specificity for SLE
- uncommon in other diseases and normal individuals
- coarsely speckled pattern with the IIF
- other methods →EIA, immunoblotting and immune diffusion

208
Q

Describe antihistone, anti-DNP, anti-ss-ARo, anti-ss-B/la and anti-nRNP tests for specific antinuclear antibodies

A

-low in sensitivity and specificity for SLE
- not used for diagnosing SLE but other tissue diseases
- Other methods →IIF,EIA, immunoblotting, and immunodiffusion

209
Q

Describe extractable nuclear antigen (ena) antibodies tests for specific anti-nuclear antibodies

A
  • Low sensitivity for SLE
    -methods that can use→ immunOdiffusion, ouchterlony double diffusion
  • typically test for the antibodies to SM
  • can be SSA, SSB and RNP
  • precipitation of lines of identity and non identity are reviewed
    -new method→ multiplex bead assay
    →use specific antigen coated beads and flow cytometry to detect multiple ANAs simultaneously
210
Q

What are serological tests for rheumatoid arthritis?

A

① rheumatoid factor
② anti-cyclic citrulinated peptide antibody (anti-ccp)

211
Q

Describe rheumatoid factor serological test For RA

A
  • Common methods → agglutination, ELISA, and nephlometry
  • autoantibody→ IgM Against the IgG
  • positive in 70-80 % of patients with RA
  • not specific for RA
  • its present with other autoimmune diseases and infections and in some normal individuals
    -agglutination only detects IgM RF but ELISA and nephlometry can also detect IgA and IgG classes of RF
    -can be automated
212
Q

Describe anti-cyclic citrulinated peptide antibody (Anti-CCP)

A

-most common method → ELISA
- more specific for RA than the RF test
- associated with move severe form of rheumatoid arthritis

213
Q

What are 4 interpretations of serological tests?

A

① greater than or equal to 4 fold increase in titer
②IgM antibody
③IgG antibody
④IgG antibody in newborn

214
Q

Describe it you see greater than or equal to 4-fold increase in titer

A
  • From acute to convalescent speciemen drawn 10-14 days later is diagnostic of disease
  • diagnostic of a disease process happening
215
Q

Describe if you see IgM antibodies in serological testing

A
  • Sign of recent infection
216
Q

Describe if you see IgG antibodies in serological testing

A
  • Sign of immunity
217
Q

Describe if you see IgG antibodies in a newborn: in serological testing

A
  • Material antibody

MLT Semester #1 (15 decks)

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