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Transfusion-Transmisible Infections, RAPID PLASMA REAGIN (RPR)
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RAPID PLASMA REAGIN (RPR)
- Macroscopic non-treponemal serological test
- cardiolipin-containing antigen suspension bound to charcoal particle with addition of EDTA, thimerosal, and choline chloride
procedure:
- Place 50 µL of serum or plasma onto a 18-mm circle of the RPR test card, using
a disposable Dispenstir or a safety pipetting device.
- Using the inverted Dispenstir (closed end) or flat toothpicks, spread the serum or
plasma to fill the entire circle.
- Gently shake the antigen dispensing bottle to resuspend the particles.
- Holding the dispensing bottle and needle in a vertical position, dispense several drops to clear the needle of air. Then add exactly 1 free-falling drop (17 µL) of antigen suspension to each circle containing serum or plasma. Do not mix
- Place the card on the mechanical rotator under a humidifying cover. Rotate the
card for 8 minutes at 100 ± 2 rpm.
VENEREAL DISEASE RESEARCH LABORATORIES
(VDRL)
- Microscopic slide flocculation test for serum that includes modification for use of CSF
- Antigen: cardiolipin, cholesterol, and lecithin
Procedure:
- Serum specimen is heated at 56 degree celsius for 30 mins to inactivate complement.
- Place 50 µL of serum or plasma onto a ceramic ring of a glass slide.
- Add 1 drop of VDRL antigen to the ring
- Place the card on the mechanical rotator under a humidifying cover. Rotate the
card for 4 minutes at 180 ± 2 rpm
FLUORESCENT TREPONEMAL ANTIBODY ABSORPTION (FTA-ABS)
- Indirect immunofluorescent, using
antigen-antibody complex
- gold standard diagnostic test for Syphilis
Procedure:
- A dilution of heat-inactivated pxn serum is incubated with a sorbent consisting an extract of Reiter strain to remove Ab that cross-react with treponemes other than T. pallidum.
- Diluted pxn sample and control are applied to individual wells on a test slide fixed with Nichols Strain of T. pallidum 3. incubated for 30 mins in 37 degree, wash and air dry
- Add Anti-human immunoglobulin conjugated with fluorescence to the well, re-incubate and wash.
- Mount and and examine under fluorescence microscope.
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a. Identify any blood products previously donated by a donor currently testing positive
b. identify all blood products previously donated by a donor 12 months before the last negative screening test.
c. Notify facilities that received units involved in the look-back investigation.
d. trace to patients and notify patients of potential exposure
LOOK-BACK and RECIPIENT FOLLOW UP
- if a patient develop HBV, HCV, HIV, or HTLV after receiving a single unit from one donor, that donor is permanently deferred.
- it the recipient received donations from several donors, all donors do not have to be excluded.
- donors may
be called in for retesting.
- If a donor has been implicated in more than one case of TTD, this donor should be retested and possibly permanently deferred.
- other recipients of a component from
the suspected donor should be contacted.
- donor must be placed on the appropriate donor deferral list if subsequent test are positive
- Notification and a thorough explanation of the positive test
results and their implications must be given to the donor
- Follow-up testing should be performed by the donor’s own
physician
RAPID ANTIGEN DETECTION SYSTEMS (RDTS)
- Based on immunochromatographic antigen detection, that detects soluble proteins through their ability to bind to capture antibodies contained in nitrocellulose strip.
Procedure:
- Add 5ul of whole blood and placed on the strip and eluded by adding a 2 drops of buffer that contains a labeled antibody. A colored band representing the antigen-antibody complex can then be seen.
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(ELISA) ENZYME-LINKED IMMUNOSORBENT ASSAY
- These tests employ a sandwich ELISA or CLIA in which patient serum is incubated with a solid phase onto which synthetic or recombinant HIV-1 antigens, HIV-2 antigens, and a monoclonal antibody to HIV-1 p24 have been attached.
- If antibody to HIV is in the sample, it will bind to HIV-1 or HIV-2 antigens, and if p24 antigen is in the sample, it will bind to anti-p24 on the solid phase.
WESTERN BLOT (IMMUNOBLOT)
- Western Blot Kits are prepared commercially as nitrocellulose or nylon strips containing individual HIV proteins that have been separated by polyacrylamide gel electrophoresis and blotted onto the test membrane.
Procedure:
*Patients serum is use for the test. During incubation period, any HIV antibodies present in the sample will bind to their corresponding antigens on the test membrane. Unbound antibody then is removed by washing.
*Next, an anti-human immunoglobulin with enzyme label is added directly to the test strip and binds to specific HIV antibodies from the patient sample.
*Unbound conjugate is removed by washing, whereas bound conjugate is detected after adding appropriate substrate, which produces chromogenic reaction.
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