HIV/AIDS testing

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Questions and Answers

HIV

Type: retrovirus, has RNA genome and RNA dependent DNA polymerase termed reverse transcriptase Pathogenesis: targets immune system by forming amounts of progeny virus in vivo Transmission: body fluids (sexual activity, blood transfusion, IV injection) Appear: surrounded by a lipid bilayer from host cell membrane during budding and a protein capsid with 2 identical RNA molecules Contains: proteins and glycoproteins Early Stages: viral reverse transcriptase has an error state, leading to high mutation rates that allow for viral variants to escape immune surveillance Diagnosis: saliva samples or serum to find presence of HIV antibodies

T cells in HIV

T cells: lymphocytes that play a role in adaptive immunity, helper release cytokines and cytotoxic kill directly.

HIV Pathogenesis

destroys TH cells by binding to their CD4 receptor , which needs a CCR5 and CXCR4 coreceptor. Viral RNA enters the cell and fuses cell membrane with viral envelope. RNA becomes DNA and it integrated in cells and makes new viral proteins, leading to cell death, leading to proteins being released that can affect other cells

HIV ELISA components

<p>Primary antibody: recognizes area of interest,recognizes one of virions coat proteins Secondary antibody: recognizes the primary antibody Horseradish Peroxidase (HRP): secondary antibody that can help detect antibody-antigen complex Antigens: viral proteins added to the wells ahead of time that can absorb to the surface</p>
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HIV ELISA steps

<p>Step 1: antigens (viral proteins) added to coat the well, washed to remove excess Step 2: pt sample added,if anti-HIV-1 IgG antibodies present they will recognize and bind to the HIV antigens. Wash to remove excess that didn’t bind Step 3: secondary antibody (HRP): is added to the well and binds to the primary antibody of its present. Wash away excess that did not bind Step 4: ABTS and hydrogen peroxide are added to each well. HRP oxidizes ABTS on wells, turing clear solution green</p>
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AIDS

<p>From: HIV infection Symptoms: Skin lesions, viral induced skin cancers, respiratory infections, immunocompromised HIV hallmarks: RNA virus, attacks T cells, spread by bodily fluids</p>
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AIDS testing

<p>AIDS Testing Purpose: to detect samples with HIV coating antigens Completed on: Test done on HIV antibody test strip</p>
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ELISA AIDS steps

<p>Step 1: antigen is added an absorbed into the wells (wash to remove excess) Step 2: Blocking substance (ex:milk protein) is added to block sites not occupied by the antigen Step 3: Serum containing antibody IgG is added , if antibodies for specific antigen are present they will bind to antigen and be absorbed into the wells Step 4: secondary antibody (Anti-IgG) is added to well and binds to primary antibody if present. Cross linked to HRP enzyme Step 5: colorless substrate is added to each well to detect antigen-antibody complex, turns pink if present</p>
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Blocking

<p>Prevents other proteins from binding to the plate</p>
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