The Leishmania IgG/IgM Rapid Test is a lateral flow chromatographic immunoassay. The test cassette consists of: 1) a burgundy colored conjugate pad containing recombinant L. donovani specific antigen RK-39 conjugated with colloid gold (Leishmania conjugates), 2) a nitrocellulose membrane strip containing a test band (T band) and a control band (C band). The T band is pre-coated with recombined antigen RK-39 and the C band is pre-coated with chicken anti-protein A. 
When an adequate volume of test specimen is dispensed onto the sample pad of the test strip, the specimen migrates by capillary action across the strip.  Anti- L. donovani Ab if present in the specimen will bind to the RK-39 antigen conjugates. The immunocomplex is then captured on the membrane by the pre-coated recombined antigen RK-39, forming a burgundy colored T band, indicating a L. donovani Ab positive test result. 
Absence of the T band suggests a negative result. The test contains an internal control (C band) which should exhibit a burgundy colored band of the immunocomplex of antibody- chicken anti-protein A -gold conjugate regardless of the color development on the C band. Otherwise, the test result is invalid and the specimen must be retested with another strip.

• 1. Bring the pouch to room temperature before opening it. Remove the test device from the sealed pouch and use it as soon as possible. 
  2. Place the test device on a clean and level surface. For Whole Blood, Serum or Plasma specimens: Hold the dropper vertically and transfer 2 drops of specimen (or approximately 50 µL) to the specimen well (S) of the test device, then add 1 drop of buffer for serum or plasma and 2 drops of buffer for whole blood. Start the timer. 
  For Fingerstick Whole Blood specimens: 
  To use a capillary tube: Fill the capillary tube and transfer approximately 50 µL (or 2 drops) of fingerstick whole blood specimen to the specimen well (S) of the test device, then add 2 
  drops of buffer and start the timer. 
  3. Wait for the colored line(s) to appear. Read results at 15 minutes. Do not interpret results after 30 minutes. 

IgMPositive:* The colored line in the control line region (C) appears and a colored line appears in test line region 2 (T2). The result is positive for Leishmania specific‐IgM antibodies.

IgGPositive:* The colored line in the control line region (C) appears and a colored line appears in test line region 1 (T1). The result is positive for Leishmania specific‐IgG.

IgG and IgM Positive:* The colored line in the control line region (C) appears and two colored lines should appear in test line regions 1 and 2 (T1 and T2). The color intensities of the lines do not have to match. The result is positive for IgG & IgM antibodies.

*NOTE: The intensity of the color in the test line region(s) (T1 and/or T2) will vary depending on the concentration of Leishmania antibodies in the specimen. Therefore, any shade of color in the test line region(s) (T1 and/or T2) should be considered positive.

The colored line in the control line region (C) appears. No line appears in test line regions 1 or 2 (T1 or T2).

Controlline(C) falls toappear.Insufficient buffer volume or incorrect procedural techniques are the most likely reasons for control line failure.

Review the procedure and repeat the procedure with a new test device. If the problem persists, discontinue using the test kit immediately and contact your local distributor.