Summary of Immunology Test Points for the Test Technician Exam

Summary of Immunology Test Points for Test Technician Exam 2017

Immunology test is one of the test points in the test technician exam. In order to facilitate the candidates to better review the knowledge about immunology test. The following is my knowledge about immunology testing, welcome to read.

0, immunological testing techniques are based on antigen-antibody reactions.

1, immunity: a physiological function of the body to recognize and reject antigenic foreign substances

2, immune defense (external); immune self-stabilization (against autoimmune diseases); immune surveillance (against tumors).

3, central immune organs: bone marrow, thymus; peripheral immune organs: lymph nodes, spleen (largest), mucosa-associated lymphoid tissues

4, B cells: mediate humoral immunity by recognizing membrane immunoglobulins to bind antigens; B-cell receptor = BCR = mIg

Surface markers: membrane immunoglobulin (SmIg), Fc receptor, complement receptor, EBV receptor, and mouse erythrocyte receptor.

Mature B cells: CD19, CD20, CD21, CD22 (mIg of mature B cells are mainly mIgM and mIgD) CD5 molecules are also detected, and they can be categorized as B1 and B2 cells.

B cell function test method: hemolytic vacuole formation test (humoral immune function).

5. T cells: mediate cellular immunity. ***Same surface marker is CD3 (polychain glycoprotein); marker for helper T cells is CD4; marker for killer T cells is CD8; T cell receptor = TCR.

***Same surface marker for T cells and NK cells is CD2 (sheep erythrocyte receptor);

CD3+CD4+CD8- = helper T cells (Th)

CD3+CD4-CD8+ = Cytotoxic T cells (Tc or CTL) (T cell-mediated cytotoxicity assay)

CD4+CD25+ = Regulatory T cells (Tr or Treg)

T-cell function assay: phytohemagglutinin (PHA) cutin (CONA) stimulates T cell proliferation. Proliferation tests are: morphologic method, nuclear method.

Separation of T cell subpopulations: affinity plate binding separation method, magnetic microsphere separation method, fluorescence-activated cell separator separation method

*E wreath test is a test for counting T cells by detecting the SRBC receptor;

6, NK cells: with cell-mediated cytotoxic effect. Direct killing of target cells (tumor cells and virus-infected cells)

Surface markers: CD16 (ADCC), CD56.

Determination of human NK cell activity of the target cells are mostly used in the K562 cell line, while the determination of the activity of mouse NK cells are often used in the YAC-1 cell line.

7, phagocytes include: monocyte-phagocyte system (MPS, surface marker CD14, including pre-monocytes in the bone marrow, monocytes in the peripheral blood and macrophages in tissues) and neutrophils. (expressing MHC class II molecules)

8, human mature dendritic cells (DC) (specialized antigen-presenting function): surface markers CD1a, CD11c and CD83.

9, immunoglobulins can be divided into secretory type (sIg, mainly in body fluids, with antibody function) and membrane type (mIg, expressed on the surface of B-cells as antigen receptor, called Membrane surface immunoglobulin)

10, immunoglobulin according to the content of how much order: IgG & gt; IgA & gt; IgM & gt; IgD & gt; IgE five categories (according to the antigenicity of the constant region of the heavy chain (CH) order)

Determination of immunoglobulin content: unidirectional circular immunodiffusion, immunoturbidimetric method.

11, immunoglobulin isotype antigenic determinant clusters are located in the constant region (CH, CL)

12, antibodies are produced by plasma cells. VH and VL (highly variable regions) on the antibody molecule are antigen-binding sites.

13, IgG: the highest level of immunoglobulin in serum (IgG1 is the highest), the main antibody in blood and extracellular fluid. It is also the main antibody of the immune response again, and is the only antibody that can pass through the placenta. Most of the antibacterial and antiviral antibodies are IgG, and some autoantibodies and ultrasensitive type II antibodies are IgG, and the second antibody in the immunological test is also dominated by IgG.

14, IgA: serotypes (monomer existence) and secretory type; secretory IgA (sIgA) for the dimer, stable performance, mainly in the gastrointestinal tract, bronchial secretion, colostrum, saliva, tears, local concentration is high, is involved in mucosal localization of immunity of the main antibody.

15, IgM: pentamer, mainly in the blood, is the largest molecular weight of Ig (also known as macroglobulin). Antibodies synthesized and secreted at the earliest stage of individual development. The first antibody produced in the humoral immune response after antigenic stimulation. An increase in serum IgM level during infection indicates recent infection. If IgM is elevated in the umbilical cord blood of a newborn, it suggests an intrauterine infection.

16, IgE: monomer structure, the lowest content in normal human serum. IgE is a pro-cellular antibody, mediating type I hypersensitivity reactions. IgE is a pro-cellular antibody that mediates type I hypersensitivity reactions. It is elevated in the serum of patients with specific allergic reactions and early parasitic infections.

17, complement: a globulin with enzyme-like activity (produced by hepatocytes, macrophages); activation pathway there are three main: the classical pathway (to bind the antigen after the IgG or IgM class of antibodies as the main activator (double-stranded DNA), C1 ~ C9 all involved), the alternative pathway (pathogenic microorganisms, the cell wall components (lipopolysaccharides) direct activation of complement C3, and then complete the process of activation of C5 ~ C9), the alternative pathway (the cell wall components (lipopolysaccharides) directly activate complement C3, and then complete the process of activation of C5 ~ C9). C9 activation process), MBL pathway (activation initiated by mannose-binding lectin (MBL) produced during the acute inflammatory phase after binding to the pathogen).

18, The complement system is most abundant in C3 and least abundant in C2.

19, inactivation: heating 56 ℃ for 30 minutes, complement inactivation

20, complement clearance of immune complexes: phagocytosis conditioning; immune adhesion; immune complex inhibition.

21, complete antigen = reactogenicity + immunogenicity. Semi-antigen = reactive origin (no immunogenicity)

22, antigen-antibody binding force (intermolecular gravity): electrostatic attraction, van der Waals force, hydrogen bonding, hydrophobic force (the strongest)

23, antigen-antibody reaction of the four major characteristics: specificity, reversibility, proportionality, stage; by the reaction conditions (such as temperature, pH, electrolytes, antigen-antibody ratio, etc.) of the impact.

24, antibody excess? Anterior band; antigen excess? Posterior band (hook effect)

25, antigen-antibody reaction can be divided into two stages: the first stage for the antigen and antibody specific binding stage, this stage of the reaction is fast, only a few seconds to a few minutes, but does not appear to be visible; the second stage for the visible reaction stage, this stage of the reaction is slow, and often takes several minutes to several hours.

26, granular antigen agglutination reaction. Soluble antigen precipitation reaction. Monovalent antigen binds to the corresponding antibody without precipitation.

27, papain hydrolyzed IgG for 2Fab + Fc; pepsin hydrolyzed IgG for F(ab)2 + nFc.

28, the most commonly used adjuvant for immunized animals is Fuchs' adjuvant, which is divided into two types of Fuchs' complete adjuvant (Fuchs' incomplete adjuvant plus BCG) and Fuchs' incomplete adjuvant.

29, R (rabbit) type antiserum is the use of rabbits and other animals immunized with antibodies, antigen-antibody reaction ratio appropriate range is wide, suitable for diagnostic reagents.

H (horse) type antisera are antibodies obtained by immunization of large animals such as horses, with a narrower range of antigen-antibody reaction ratio, generally used as immunotherapy.

30, antisera common preservation conditions: 2-8 ℃ preservation; frozen preservation (most commonly used); vacuum dry preservation (5-10 years).

31, hybridoma cells into liquid nitrogen (-196 ℃) before the need for gradual cooling. When resuscitating the cells, remove the cryotubes from the liquid nitrogen tank and immediately immerse them in a 37°C water bath.

32, the agglutination reaction is divided into two stages: ① specific binding of antigen and antibody; ② the appearance of visible particle coagulation.

33, the principle of direct agglutination reaction is bacteria, spirochetes and erythrocytes and other particulate antigens, in the appropriate electrolyte (0.85% sodium chloride) can be directly with the corresponding antibody binding agglutination. Antigen = agglutinogen, antibody = agglutinin.

34, slide agglutination test: mainly used for qualitative analysis of antigen, AB0 blood group determination.

In vitro agglutination test: Fertilizer test, Exophthalmos test, transfusion cross-matching test;

35, erythrocyte encapsulated antigen, used to detect the antibody hemagglutination reaction, known as the positive indirect hemagglutination reaction (PHA).

Red blood cells covered with antibodies to detect antigen hemagglutination reaction, known as reverse indirect hemagglutination reaction (RPHA).

36, indirect hemagglutination inhibition test: can be used to detect antibodies, autoantibodies, allergic antibodies, but also determine the antigen.

37, Staphylococcus aureus A protein (SPA)

38, hemagglutination test can be carried out in the microtitre plate or test tube, the specimen will be doubly diluted, usually 1:64, while setting up a dilution without specimen as a control hole. Where erythrocytes are deposited at the bottom of the well, concentrated in a dot for non-agglutination. If the erythrocytes are agglutinated, they are distributed around the bottom of the hole.

39, gelatin agglutination test (indirect): HIV-1 antibody and anti-sperm antibody detection

40, antiglobulin test, also known as the Coombs test, is a very useful method of detecting incomplete anti-erythrocyte antibodies. Including direct Coombs test (detection of incomplete antibodies on red blood cells) and indirect Coombs test (incomplete antibodies free in the serum)

41, precipitation reaction in two stages: the first stage for the antigen and antibody specific binding, a few seconds to tens of seconds can be completed, the emergence of small soluble complexes, invisible to the naked eye. The second stage is the formation of visible immune complexes, it takes about tens of minutes to hours to complete, such as precipitation line, precipitation ring.

42, immunoturbidimetric: optimal pH 6.5 to 8.5, phosphate buffer.

43, convection immunoelectrophoresis: antibody flow to the negative pole, antigen flow to the positive pole

44, immunofixation electrophoresis is also used for the detection of this - weekly protein in urine and ?

45, RIA (radioimmunity) nuclide (125 Ⅰ) labeled antigen, competitive inhibition (labeled antigen and non-labeled antigen competition limited antibody); IRMA (immunoradiotherapy) nuclide labeled antibody, non-competitive binding (excess labeled antibody, the reaction rate faster than the RIA, the sensitivity is significantly higher than that of the RIA.)

RIA can be determined by the determination of large molecules and small molecules of antigen, but IRMA can only determine at least one of the antigens. IRMA, on the other hand, can only determine antigens with at least two antigenic determinants.

46, commonly used fluorescein: isothiocyanic acid (FITC) yellow-green, the most commonly used; tetraethyl rhodamine (RB200) orange-red; tetramethyl rhodamine isothiocyanate (TRITC) orange-red; phaeohydrococcus (R-RE) orange. Others: europium (Eu3+)

47, the common methods of fluorescence labeling protein: stirring method and dialysis method.

Fluorescent antibody potency identification: when the antigen content is 1g/L, the antibody potency >1:16

48, Time-resolved fluorescence immunoassay (TRFIA) uses lanthanide (e.g., europium) compounds as fluorescent markers.

49, Polarization immunoassay polarization wavelength is 485nm (blue light).

50, Horseradish peroxidase (HRP): substrate (1) o-phenylenediamine (OPD), (2) tetramethylbenzidine (TMB) The most widely used substrate in ELISA.

Alkaline phosphatase (ALP): substrate: p-nitrophenyl phosphate (p-NPP)

? -Galactosidase (? -Gal): substrate: 4-methylumbelliferyl-R-D galactoside (4-MUU)

51, Heterogeneous method: separation first, then measurement; homogeneous method: no separation, direct measurement.

52, enzyme-linked immunosorbent assay (ELISA):

Necessary reagents: ① solid-phase antigen or antibody; ② enzyme-labeled antigen or antibody; ③ substrate for enzyme reaction.

Antigen Determination: Protein macromolecular antigens are most often used double antibody sandwich method (HBsAg). Small molecules with only a single antigenic determinant cluster use the competitive inhibition method.

Antibody assays: Indirect (HCV, HIV), double antigen sandwich (HBsAb), competitive (HBcAb, HBeAb), and capture (IgM) assays are commonly used

*The last color shade displayed in the wells to be tested correlates positively with the antigen or antibody to be tested in the specimen: double-antibody sandwich, two-site one-step method, and indirect method of measuring antibodies;<

53, chemiluminescent substrates are: direct chemiluminescent agent: acridine ester and terpyridine ruthenium; enzyme reaction luminescent agent: (labeled enzyme HRP) luminal and its derivatives, (labeled enzyme alkaline phosphatase) AMPPD;

54, immunoblotting experiments are often used in the selection of polyclonal antibodies.

55, the separation of peripheral blood single nucleated cells of the layering fluid commonly used: Ficoll (from top to bottom for plasma, single nucleated cells, granules, red) and Percoll (from top to bottom: dead cells, single nucleated cells, gonorrhea, red, granules).

56, the collection of pure lymphocyte populations are: adhesion to the wall method, adsorption column filtration, magnet attraction method, Percoll separating fluid method.

57, the separation of T cells and B cells: E wreath sedimentation method, nylon hair column separation method

58, the determination of lymphocyte viability: Tepan blue staining method, dead cells are blue.

59, CD4 is the HIV receptor, and the CD4/CD8 ratio is significantly reduced in HIV infection.

60, CD4/CD8 elevation is common in autoimmune diseases; and CD4/CD8 decrease is common in viral infections, malignant tumors and aplastic anemia.

61, (ELISA) double antibody sandwich method is the most commonly used method for cytokine determination.

62, flow cytometry: forward scattered light (FS) reflects the size of the particles. Side-scattered light (SS) reflects the complexity of the internal structure of the particle, the smoothness of the surface. Fluorescence (FL) reflects how much of the particle is colored with fluorescence.

63, immunofluorescence labeling of the most commonly used fluorescent dyes: fluorescein isothiocyanate (FITC) bright green fluorescence. Texas red red fluorescence. Algae bile protein class orange to red fluorescence.

64, Tricolor fluorescent antibody labeling is commonly used in clinical practice to identify CD3-CD16+CD56+ lymphocytes as NK cells.

65, A characteristic immunodiagnostic indicator of AIDS patients is manifested as a decrease in the total number of T lymphocytes, an inverted ratio of the T-cell subpopulation CD4Th/CD8Tc, Th/Tc<1.0, and a significant decrease in the number of Th cells or even undetectable, whereas the number of Tc cells can be normal or increased, the number of NK cells decreases or decreases in their viability, and the B-lymphocyte population is in the normal range.

65, ankylosing spondylitis?HLA-B27

66, SLE patients with IgG, IgA elevation is more common. Patients with rheumatoid arthritis are predominantly characterized by increased IgM.

67, M protein (MP) is an abnormal monoclonal immunoglobulin produced by monoclonal abnormal proliferation of B lymphocytes or plasma cells (>30g/L), which is very homogeneous in amino acid composition and sequence. Most of them are not immunologically active, so they are also called paraproteins. Clinically seen in multiple myeloma, hypergammaglobulinemia, malignant lymphoma, heavy chain disease, light chain disease, etc.

68, M protein - the most basic method: serum protein zonal electrophoresis technology

M protein - rough screening test: serum immunoglobulin quantification (one-way agar immunodiffusion and immunoturbidimetric method)

M protein - identification, the preferred method: Immunofixation electrophoresis (IFE) technique (zone electrophoresis technique combined with an immunoprecipitation reaction with a specific antiserum) is the most commonly used clinical method.

69, Decreased IgD is seen in primary apheresis, silicosis

70, Concentrations in CSF: IgG>IgA>IgM. In neurological tumors, IgA and IgM are predominantly elevated.

(Infections, vascular lesions, systemic diseases = elevated IgG)

71, This - periplasmic protein is the free immunoglobulin light chain in urine. It can be detected in urine, but negative in blood (the reason is that the small molecular weight of this periplasmic protein, it is very easy to be rapidly excreted from the kidneys, and the level in blood is not elevated)

72, cryoglobulin, also known as cold immunoglobulin, precipitation occurs at -4 ℃, and then re-dissolved at 37 ℃. Blood collection is the key to the detection of cryoglobulin, and it is advisable to collect and preserve venous blood at body temperature.

73, Complement binding test (CFT), diagnosis of syphilis, Wahl reaction.

74, Nonhomogeneous fluorescence immunoassay: time-resolved fluorescence immunoassay.

Homogeneous fluorescence immunoassay: fluorescence polarization immunoassay.

75, Streptococcal infection: ASO (latex agglutination test, immunoscattering turbidimetric assay)

76, Salmonella typhi has a mycobacterial (O) antigen, flagellar (H) antigen and surface (Vi) antigen. Fertilizer reaction, potency ?1:80 for positive

77, Pneumocystis carinii, also known as Pneumocystis carinii, is a fungus.

78, Type I hypersensitivity reactions: mediated by specific IgE antibodies, which occur most rapidly. (Penicillin, bronchial asthma)

Increasing the activity of Th1 cells and decreasing the secretion of IL-4 reduces the production of IgE and blocks the IgE-mediated type I hypersensitivity reaction.

Involved in type I hypersensitivity reaction: mast cells, basophils, eosinophils.

79, Type II hypersensitivity reaction: also known as cytotoxic or cytolytic hypersensitivity reaction, which is caused by the combination of antigen on the surface of the target cell with the corresponding IgG or IgM class antibody, in the complement (cytolysis), macrophage (phagocytosis) and NK cells (ADCC effect) involved in the pathological immune response with cell lysis or tissue damage. (Blood transfusion reaction, neonatal hemolytic disease, autoimmune hemolytic anemia, drug-allergic hemocytopenia, pulmonary hemorrhagic nephritis syndrome, hyperthyroidism)

80, type Ⅲ hypersensitivity reaction: soluble immune complexes deposited in the basement membrane of the local or systemic capillaries, through the activation of the complement, and in the platelets, basophils, neutrophils, etc., caused by congestion and edema, localized and systemic. Inflammatory reaction and tissue damage mainly characterized by congestion and edema, local necrosis and neutrophil infiltration. (Arthus reaction, Arthus-like reaction, serum sickness, post-streptococcal infection glomerulonephritis, rheumatoid arthritis, SLE)/81, Type IV hypersensitivity reaction: also known as delayed-type hypersensitivity reaction (DTH), is an inflammatory reaction mainly characterized by infiltration of single nucleated cells and tissue damage caused by the combination of effector T cells and specific antigen. (Cellular immunity) Activation of effector CD4+ Th1 cells upon recognition of antigen results in the release of IFN-? , TNF, lymphotoxin (LT), IL-3, GM-CSF, monocyte chemotactic protein-1 (MCP-1) and other cytokines.

Common type IV hypersensitivity disorders: infectious delayed hypersensitivity (Mycobacterium tuberculosis, viruses, protozoa), contact dermatitis, transplant rejection. (Tuberculin skin test, patch test)

82, denatured IgG can stimulate the body to produce anti-denatured IgG antibodies (rheumatoid factor)

83, ITP patients have anti-platelet antibodies in the serum, which can shorten the life span of platelets.

84, Anti-acetylcholine receptor antibodies: diagnostic for myasthenia gravis (MG).

85, Patients with toxic diffuse goiter have IgG-type autoantibodies against thyrotropin hormone receptor (TSHR) in their serum. (The 'hyperthyroidism)

86, polymyositis is an autoimmune disease that primarily manifests itself by damage to the muscles, and if there is also skin damage, it is called dermatomyositis.

87, 75% of patients with PSS have positive antinuclear antibodies, anti-Scl-70 antibodies are specific for PSS, and 80% to 95% of patients with limited scleroderma are positive for anti-binding point antibodies.

88, autoimmune disease (AID) features: high titers of autoantibodies, pathology is characterized by immunoinflammatory damage consistent with the distribution of antigens, and can build animal models.

89, Anti-DNP antibodies (anti-nucleoprotein antibodies) are usually completely taken up by DNA and histones and are factors in the formation of lupus cells.

90, ANA positive fluorescence phenomenon is divided into: nuclear membrane type, homogeneous type, speckled type, kernel type.ANA is often used as a primary screening test for autoimmune diseases.

91, ENA is a general term for extractable nuclear antigen, which does not contain DNA in the molecule.

92, anti-dsDNA antibody has a high specificity for SLE. Anti-Sm antibodies are also one of the specific markers of SLE. These two items are often indicators of a confirmed diagnosis of SLE. (In patients suspected of having SLE, ANA and anti-dsDNA antibodies should be tested first, and when ANA and/or anti-dsDNA antibodies are positive, then the anti-ENA antibody profile should be tested. If anti-Sm antibody and/or anti-RNP antibody is positive, laboratory diagnosis of SLE can be made)

93, anti-nuclear RNP antibody: a diagnostic indicator of MCTD (mixed connective tissue disease).

94, anti-SSA/anti-SSB antibodies: a diagnostic indicator for dry syndrome (SS). (Laboratory diagnosis of dry syndrome can be made when ANA is positive and anti-dsDNA antibody is negative, and anti-SsA antibody and/or anti-SSB antibody is positive in the anti-ENA antibody spectrum.)

95, anti-Jo-1 antibody: patients with polymyositis (PM) are often positive.

96, Anti-Scl-70 antibody: diagnostic indicator of progressive systemic scleroderma (PSS).

97, NCA: a specific serum marker for primary small vessel vasculitis

98, endocrine rheumatoid arthritis (RA): autoantibodies are RF, antikeratin antibody (AKA), and anti-cyclic citrullinated peptide antibody (anti-CCP).

99. The Farr method for determining dsDNA antibodies has high specificity and is recognized as a standard method, which is meaningful for SLE when the anti-dsDNA antibody binding rate is greater than 20%.

100, anti-mitochondrial antibody (AMA): primary biliary cirrhosis

101, macroglobulinemia: a disease with malignant proliferation of IgM-secreting plasma cells as the basis of pathology. Prevalent in elderly men, the main manifestations of extra-marrow infiltration, with liver, spleen and lymph node enlargement as the main signs and accompanied by hyperviscosity syndrome, such as manifestations of retinal hemorrhage. (The serum is jelly-like and difficult to separate, and the serum is sometimes difficult to swim during electrophoresis, focusing on the origin is the electrophoretic characteristics of the disease)

102, abnormal immunoglobulin detection of the application of the principle

Initial screening experiments: zonal electrophoresis analysis, quantitative immunoglobulin detection and qualitative detection of the urine Ben - peripheral protein.

Confirmatory experiments: immunoelectrophoresis, immunofixation electrophoresis, immunoglobulin subtyping, quantitative detection of serum and urinary light chain protein.

103, HIV-induced immune response

1. humoral immune response after HIV infection, the body can produce: neutralizing antibodies, anti-P24 chitin antibodies, anti-gp120 and anti-gp41 antibodies

2. cellular immune response: CD8 + T cell response, CD4 + T cell response.

104, HIV antigen detection: core antigen p24 (ELISA)

105, immunoblotting judgment criteria are: ① HIV antibody positive: at least two membrane bands (gp41/gp120/gp160) or at least one membrane band and the p24 band at the same time; ② HIV antibody negative: no HIV antibody-specific bands; ③. HIV antibody suspicious: the appearance of HIV-specific bands, but the band type is not enough to confirm the positive person.

CD4+ T-cell count is the clearest indicator of the state of immune system damage in HIV-infected patients. When CD4+ T cells are lower than 500/?l, it is easy to opportunistic infections; lower than 200/?l, AIDS occurs.

106, CEA is greater than 60?g/L can be seen in colon, stomach, lung cancer. CEA levels are normalized 6 weeks after surgery.

107, AFP?primary liver cancer, PSA?prostate cancer

108, Glycoantigens: CA125: epithelial ovarian cancer and endometrial cancer; CA15-3: breast cancer; CA19-9: pancreatic cancer, colorectal cancer.

109, neuroblastoma and small cell lung cancer (gene has P53, RB) markers: neuron-specific enolase (NSE)

110, rejection reactions are: host-versus-graft reaction (HVGR) and graft-versus-host reaction (GVHR).

111, the relationship between graft survival and HLA mating is: ① the more donor and recipient HLA-A and HLA-B matching loci, the higher the chance of graft survival; ② donor and recipient HLA-DR loci matching is more important, because the HLA-DR and HLA-DQ genes have a strong chain of imbalance, the DR loci matching individuals, usually the DQ loci are also matched; ③ The relationship between the degree of HLA matching and transplantation outcome in different regions has different predictive values.

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