A HIGHLY SENSITIVE AND RAPID PROCEDURE FOR DIRECT PCR DETECTION OF LEISHMANIA-INFANTUM WITHIN HUMAN PERIPHERAL-BLOOD MONONUCLEAR-CELLS

We have developed a highly sensitive, simple and rapid procedure to de tect Leishmania infantum within human macrophages. It only requires fi coll preparation of peripheral blood mononuclear cells from the patien t, and their direct use for Leishmania kDNA amplification by polymeras e chain reaction. Under these conditions, about one parasite can be de tected in a one million human cell environment. Results, including tho se of a hybridization step to confirm the diagnosis specificity, are o btained within 24 h, a very short period as compared to current diagno stic methods. This procedure is of particular interest for early detec tion and early drug treatment of leishmaniasis, especially in the case of HIV coinfection. Furthermore, the method could be useful for monit oring the efficiency of new leishmaniasis treatments in infected patie nts.