Research Reagent for the Determination of Reticulated Platelets and Absolute Count Determination of Thrombocytes
- Simple method for the flow cytometric determination of reticulated platelets.
- Absolute count determination of thrombocytes at low level counts in a combined measurement.
- Standardized test procedure
- Reagents for 50 tests.
Test kit contains CD42b RPE conjugate, RNA/DNA staining solution, dilution buffer and counting tubes.Reagents für 50 tests. Test kit contains CD42b RPE conjugate, RNA/DNA staining solution, dilution buffer and counting tubes. Evaluation by flow cytometry.
SUMMARY and EXPLANATION
THROMBOCYTEST® plus allows the flow cytometric determination of reticulated platelets and the absolute count determination of thrombocytes during thrombocytopenia (number of thrombocytes < 20.000/µl). The test kit contains a RNA specific dye which stains the RNA from reticulated platelets, a monoclonal antithrombo-cyte antibody conjugated with the fluorescence dye phycoerythrin (PE) and fluorescent beads as counting standard.
APPLICATIONS
Reticulated platelets are young platelets that contain increased levels of mRNA and rRNA compared to mature cells. When thrombocytes are released from the bone marrow into the peripheral circulation they contain residual RNA which is subsequently degraded as the cells circulate. These young reticulated platelets appear normally in the peripheral blood at low levels, up to 4.5% of total thrombocytes.
An increased proportion of reticulated platelets indicates increased thrombopoiesis. The ability to detect increased platelet production has proven to be useful clinically in patients with thrombocytopenia. If these patients have elevated levels of reticulated platelets it implies that they have a disease resulting in peripheral destruction of platelets. In contrast, if their levels of reticulated platelets are depressed, it implies that they have a disease which impairs the ability of the bone marrow to make new platelets.
The determination of reticulated platelets is useful for the evaluation of thrombopoietic disorders like thrombocytopenia, for monitoring course and treatment of Idiopathic Thrombocytopenic Purpura (ITP) (see Fig. 4), of bone marrow transplantation success und during chemotherapy. Platelet-associated immunoglobulin (PA-Ig) which has been correlated with ITP can be detected by the separate kit THROMBOCYTEST® immune.
Also, the effect of drugs on the rate of platelet production can be studied.
TEST PRINCIPLES
THROMBOCYTEST® plus allows the flow cytometric determination of reticulated platelets as well as the determination of absolute counts of thrombocytes. The test kit contains a specific RNA dye which stains the RNA from reticulated platelets, a monoclonal antibody against a thrombocyte specific antigen conjugated with the fluorescence dye phycoerythrin (PE) and fluorescent beads as counting standard for absolute counting of thrombocytes.
The anti-thrombocyte antibody (“CD42b RPE Conjugate”) is added into a test tube which contains counting beads (“COUNTING TUBES”) or into a test tube without counting beads, if only the percentage of reticulated platelets has to be determined. Then, EDTA whole blood and a RNA specific dye (“RNA/DNA STAINING SOLUTION”) is added. After an incubation period of 30 min at room temperature the cell suspension can be analyzed by flow cytometry.
REAGENTS PROVIDED
- CD42b RPE Conjugate
- RNA/DNA STAINING SOLUTION
- DILUTION BUFFER
- 10 COUNTING TUBES
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