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HIV TESTING
Screening assays
DIAGNOSIS
Tests to Confirm HIV Infection
Who?
Mandatory Muslim brides Prisoners-drug addicts Entrance test-Pusat Serenti Recommended pregnant women Persons at high risk for HIV infection
Where?
Available in all government clinics and hospitals FOR FREE Available in private clinics and hospitals.
Immunologic Principle
particle agglutination, immunodot (dipstick), immunofiltration (flow-through device), immunochromatography (lateralflow)
Rapid Test
Rapid Test
Rapid Test
Advantage Good for testing 1 to 100 specimens at a time Requires minimal equipment and reagents Can be performed in a clinic (onsite testing) Highly skilled staff not required Very easy to interpret test results Results in < 45 minutes Test kits can be stored at room temperature (increased stability)
Limitations Not good for testing >100 specimens at a time The QA/QC is performed at multiple sites: requires more control May cost more perindividual test than EIA Choice of testing strategy may require multiple specimens Interreader variability may provide inconsistent results with some assay formats (e.g.,particle agglutination)
Methodology
-electrophoretic technique to separate HIV antigens derived from a lysate of virus grown in culture -("blotted") to nitrocellulose paper -paper is cut into thin strips (each with the full distribution of viral protein antigen bands) -incubated with a 1:50 or 1:100 dilution of a test sample or a control -Washed~incubated with a labeled (tagged) antihuman globulin -enzyme (horseradish peroxidase or alkaline phosphatase) react with antigen-antibody complex -band formed
Positive
Negative
Indeterminate
Positive
For positive classification CDC guidelines-which require reactivity to at least 2 of the following antigens p24, gp41, gp120/160
Negative
For negative classification Absence of all bands Exception for WHO -that results also can be reported as negative if there is only a very weak p17 band
Indeterminate
For indeterminate classification -is reactivity to 1 or more antigens, but not fulfilling the criteria for positivity -indeterminate results show only weak reactions to the Gag proteins (mostly p17, p24 and/or p55)
Indeterminate WB result
- WHO recommends retesting persons after 2 weeks, -while other organizations suggest waiting 1-6 months before retesting
Retest result
progress serologically (more bands or greater intensity of bands) or converts to positive (seroconversion)
* individuals who have received vaccination for HIV (eg, subunit gp160) may be misidentified as positive based on reactions to the envelope antigens alone
Cause
hypergammaglobulinemia, the presence of cross-reactive antibodies, infection by HIV-2, infection by an unknown (but related retrovirus)
* autoimmune diseases (eg, systemic lupus erythematosus) can cause falsepositive HIV tests
Limitations
Nonquantitative Training It is complex to administer and may produce indeterminate results if a person has a transitory infection with another virus.
the initial and second tests must be of different principle (bead vs microtiter) and/or use a different antigen source (lysate vs recombinant or synthetic peptide).
inconsistent results
Due-mislabeling or technical errors.
Technical Errors
Solution -dedicated supervisory review mechanism -Outline the quality assurance measures
Solution Health care providers can be vigilant by inquiring as to the geographic origin of persons tested, or their contact with persons from these areas of Africa
Viral Load
Summary
Malaysia Rapid Test Western Blot *PCR for infants and children