Dear George, For heparin-induced thrombocytopenia (HIT) serotonin release assay, can serotonin ELISA sensitive to HIT be used? With regards, Vilas Hiremath.
Hello, Vilas, always good to hear from you. I had to go to experts, Drs Larry Brace, Edward Hospital in Naperville IL, and Jon Geske, Precision BioLogic Manager of Emerging Technologies & Markets. Here is a reply from Dr. Geske:
Theoretically, a serotonin ELISA could be used to measure serotonin released from platelets activated by antibodies from HIT patients. Unlike the gold standard 14C-serotonin release assay, however, one would have to differentiate between serotonin present in the patient’s sample vs. serotonin released from the activated platelets. Controls would have to be run (low vs. excess heparin levels) similar to the 14C assay to detect PF4/heparin antibody-dependent platelet activation. Furthermore, the ELISA method for serotonin quantification is complicated by the fact that many such assays require serotonin acylation prior to running the ELISA. Having cleared those hurdles, however, an ELISA could be used to detect serotonin levels. I hope this helps. Jon.
Hi Vilas and George. This is a thought provoking question f
Hi Vilas and George. This is a thought provoking question for the reason that if one could come up with a serotonin release assay (SRA) that could be performed using ELISA methods you could eliminate the need for all of the special handling, training and disposal of radioactive waste. In principle one would still need to incubate “donor platelets” with serotonin, perform the usual steps for a C-14 SRA (without the C-14) and then use the ELISA serotonin to pick up the released serotonin from the platelets. The last test system we used for platelet serotonin was from IBL and it included an overnight incubation. It was quite a cumbersome test. (their website shows a few shorter versions currently). The SRA is a complicated test process that demands strict attention to detail. There are current ELISA test kits that are adequate for most circumstances.
Regards, Herb Crown
St. Louis University Hospital Coagulation Lab