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ImmunoTest™ HIV-1 p24 For Serum, Plasma, Tissue Culture Medium
Strictly for non-human research and diagnostics
ImmunoTest™ HIV-1p24 test is an in vitro, semiquantitative immunochromatographic assay for the detection of p24 core protein in blood, serum/plasma, and tissue culture medium samples.
ImmunoTest™ One-Step HIV-1 p24 Test utilizes antibodies specific for the most conserved epitopes of HIV-1 core protein p24 (proprietary) in immunochromatographic assay technology for semiqualitative detection of p24 antigen in, serum, plasma or tissue culture medium. In the test procedure, 20ul of serum/ plasma or tissue culture medium is applied to the sample well (S) of a test cassette. p24 core protein binds to anti-p24 mAb gold conjugate and is then captured by the solid phase p24 specific mAb stripe on the test membrane. Visualization of the p24 core protein in the Test window will occur only when the antibody-dye conjugate binds to the antigen. As the antibody-dye conjugate continues to move along the test membrane, it will bind to another band located in the Control window (C) to generate a colored band regardless of the presence of p24 in the test sample. Therefore, the presence of two colored bands, one in the Test window and the other in the Control window, indicates the presence of p24 core protein in the test sample, while the absence of a distinct colored band in the Test window indicates a negative result.
1. Tissue culture virus 25ul + 0.05% Tween-20, heat inactivate at 55C for 5 minutes.
2. Transfer 20ul of inactivated test sample (1), or human serum/plasma to sample well (S).
3. Directly deposit 3 drops of developer from the dropper bottle to the sample port (S) in the cassette.
4. Read results in 5 - 30 minutes.
Two colored lines in the Reading windows – one each in the upper Control area and lower Test area indicate the presence of p24 core protein in the test sample. The test result can be read as soon as a distinctive pinkpurple line appears in the Test area. In most strong positive cases, the Test line will appear before the Control line. The Test line may appear after the Control line in some weak positive cases, and the Control line may become darker than the Test line. The three possible positive cases, therefore, are:
a. Two strong colored lines, one in each area - T and C .
b. One strong colored line in the Test area and one lighter colored line in the Control area.
c. One light colored line in the Test area and one strong colored line in the Control area.
Only one colored line in the Control area, with no distinctive colored line in the Test area indicates that no p24 core protein was detected.
A distinctive colored line should always appear in the Control area. The test is invalid if no line forms in the Control area.
ImmunoTest™ HIV-1 p24 core protein test is a highly specific and sensitive method for the detection of p24 core protein in serum/plasma and tissue culture medium. The test is sensitive to approximately 10pg/ml p24 (0.2pg/test) in the test sample.
Stability and Storage:
ImmunoTest™ HIV-1 p24 core protein test kit should be stored at ambient temperatures (20- 28 C) in the sealed pouch. The test kit is stable until the expiration date stamped on the pouch when stored under these conditions. Do not use the test if the pouch is damaged or the seal is broken. Keep the test in the sealed pouch until ready for use.
In commercially available HIV-1lysates (1mg/ml protein) p24 was detected at 1:100,000 dilution. Recombinant p24 was detected in <0.5pg/20ul range, comparable to the sensitivity of p24 detection by Elisa. p24 antigen positive serum conversion panel members (BBI panels) showed p24 antigen positive tests – distinct T-line, results comparable to Abbotts and Coulters FDA approved p24 Elisa test results. These results were consistent with PCR/RT-PCR test results of these conversion panels.
Using a micro-pipettor, deliver 20ul of the developer to the bottom of the p24 reference vial. Vortex gently and quantitatively transfer the mixed p24 reference to cassette sample (S) port, followed by 2 drops of the developer into the sample port (S). Positive red T-line will appear in a few minutes which will enhance significantly in 10 minutes. p24 reference positive red T-line validates p24 human serum test results.
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