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Lytic assays including flow cytometry

The complement-mediated lysis of trypomastigotes test, developed by Krettli and Brener in the 1980s,65 was used as a diagnostic tool and evaluation of treated patients. The rationale of this test was to screen for lytic antibodies, present in infected individuals, but absent in those successfully treated. This antibody would disappear before those detected by conventional tests.89 In practice it is a rather simple test that reacts alive trypomastigotes from irradiated mice with sera. If lytic antibodies are present, the parasites will be killed after the addition of complement when counted in a cytometer (Neubauer chamber). This apparently simple test has several drawbacks that made it unsuitable for routine use. Among them, the use of irradiated mice or cell lines to grow trypomastigotes poses a problem and the use of living parasites makes it potentially dangerous. The use of complement adds more problems, similar to those described with the CF test.

More recently, the adaptation of this test with flow cytometric techniques shows good results with the group of treated patients, but the test has been employed only in one research institution and is not available in the market.90-92

 
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