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Guide to the operation of the black fever rapid detection strip
Black fever rapid detection strip operation guide
(US Inbios imported original INS015/INS020/INS025)
Expected use
The test strip is used for the diagnosis of visceral leishmaniasis, and rapid detection of L. donovani antibodies in human serum by rapid immunochromatography. This experiment is helpful in diagnosing visceral leishmaniasis (VL). And only for in vitro diagnosis.
Experimental principle
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The test strips contained in the sealed bag or bottle should be stored at 20-28 ° C (room temperature) until the expiration date. The capture buffer contained in the bottle should also be stored at 20-28 °C. Exposure to temperatures above 30 °C can affect experimental performance and should be avoided. The test strip should not be stored frozen. It should be used within 1 hour after being taken out of the sealed bag to avoid being affected by humidity.
Sero collection
1 Human serum should be used in this experiment. Whole blood samples should not be used because excessive background may affect the correct reading of the experimental strip. The serum should be diluted with buffer to avoid direct experimentation. Positive sera should be diluted with negative serum.
2 After serum coagulation, the serum is separated by centrifugation to avoid hemolysis.
3 Experiments should be carried out as soon as possible after serum collection. Serum should not be stored at room temperature for long periods of time. Serum can be stored for 3 days at 2-8 °C. Otherwise the serum should be kept below 20 °C.
4 Serum was equilibrated to room temperature before the experiment. Frozen serum must be fully melted. Serum avoids repeated freezing and thawing.
5 If the serum needs to be transported, its packaging should comply with federal infectious source transport regulations.
Kit component
The nitrocellulose membrane of the kala-azar rapid detection strip coated recombinant rK39 in the experimental area, and coated chicken anti-protein A in the quality control area. The composition of the kit is as follows:
1, 25 separate test strips or 25 test strips in a desiccant containing bottle.
2, 1 bottle of capture buffer
Experimental procedure
1 Allow the serum to equilibrate to room temperature before the experiment.
2 Remove the black fever rapid test strip from the sealed bag or bottle
3 Add 20 ul of serum below the test strip arrow.
4 Place the test strip in the test tube, or in the microwell of a 96-well tissue culture plate, the end should be facing down according to the arrow on the test strip.
5 Add 2-3 drops (150 ul) of capture buffer
6 Read the results in 10 minutes. The clarity of the background is very important before reading the results. Especially when the sample is a low titer anti-Leishmania antibody, only a very weak band appears in the experimental area (T). Reading after 10 minutes may result in an erroneous result.
Note : Do not just add capture buffer for the experiment. You must first add 20 ul of serum.
If no movement of colloidal gold was observed for 10-15 seconds after the addition of the capture buffer, the sample addition zone on the strip was gently pressed until the movement of the colloidal gold was observed.
Explanation of results
Positive result
When the experiment is positive, both the quality control zone and the experimental zone appear, as shown in Figure 1. A positive result indicates detection of Leishmania donovani antibodies. A weak ribbon is also a positive result. As an explanation guide, the color intensity of the red band in the experimental area is related to the anti-Leishmania antibody concentration. The experimental band is a "weakly positive" serum sample. The experimental result may be between a weakly positive red band and a red dot, almost a white background ("weak positive" samples contain low affinity for recombinant antigen or low titer antibodies).
Negative result
If the experiment is negative, only the quality control band appears. A negative result indicates that no Leishmania donovani antibody was detected. There is no experimental band, as shown in Figure 2.
Invalid result
There is no quality control belt and no experimental belt. If the quality control band does not appear, the experiment is also ineffective, even if the experimental band appears. It is recommended to experiment again with new test strips and new serum.
Note : The change in the red band in the experimental area is dependent on the anti-Leishmania antibody concentration. However, this experiment does not detect the quantitative value of the antibody and the percentage increase.
Expected value:
In popular areas, the kala-fast rapid detection strip sensitivity is 90% or higher. The specificity may vary from region to region. For example, more than 13 of the 104 healthy people in India were positive.