Leishmania DNA is rapidly degraded following parasite death: an analysis by microscopy and real-time PCR
| Autor: | Eric Prina, Geneviève Milon, Emeric Roux, Denise Mattei |
|---|---|
| Přispěvatelé: | Immunophysiologie et Parasitisme Intracellulaire, Institut Pasteur [Paris] (IP), Biologie des Interactions Hôte-Parasite, Institut Pasteur [Paris] (IP)-Centre National de la Recherche Scientifique (CNRS), Institut Pasteur [Paris], Institut Pasteur [Paris]-Centre National de la Recherche Scientifique (CNRS) |
| Rok vydání: | 2007 |
| Předmět: |
MESH: Microscopy
030231 tropical medicine Immunology Microbiology Parasite load Polymerase Chain Reaction law.invention 03 medical and health sciences Mice 0302 clinical medicine law MESH: Drug treatment Leucine parasitic diseases Parasite hosting Animals [SDV.MP.PAR]Life Sciences [q-bio]/Microbiology and Parasitology/Parasitology Amastigote MESH: Mice Polymerase chain reaction 030304 developmental biology Leishmania 0303 health sciences Microscopy biology Cell Death Macrophages MESH: DNA Kinetoplastida MESH: Macrophages Antipruritics DNA Protozoan biology.organism_classification Virology 3. Good health Infectious Diseases Real-time polymerase chain reaction MESH: Persistence Kinetoplast MESH: Real-time PCR Female MESH: Leishmania amazonensis MESH: SYBR Green I |
| Zdroj: | Microbes and Infection Microbes and Infection, 2007, 9 (11), pp.1307-15. ⟨10.1016/j.micinf.2007.06.005⟩ Microbes and Infection, Elsevier, 2007, 9 (11), pp.1307-15. ⟨10.1016/j.micinf.2007.06.005⟩ |
| ISSN: | 1286-4579 |
| DOI: | 10.1016/j.micinf.2007.06.005⟩ |
| Popis: | Control of human leishmaniases relies on appropriate diagnosis and reliable methods for monitoring chemotherapy. The current method used for estimation of parasite burden during chemotherapy patient follow-up as well as in pharmacological studies performed in experimental models involves PCR-based assays. Compared to time-consuming conventional methods, this type of Leishmania DNA detection-based method is extremely sensitive, but could fail in distinguishing viable Leishmania from slowly degenerating ones. We have used an in vitro model to monitor the duration of Leishmania DNA persistence in mouse macrophages following exposure to l -leucine ester, a molecule otherwise known to rapidly kill intracellular Leishmania amazonensis amastigotes. At 1 h of post l -leucine ester exposure, more than 98% of amastigote-loaded macrophages harbored killed parasites and parasite remnants, as assessed by microscopy. This dramatic decrease in parasite load and the microscopic parasite follow-up over the 120 h time period studied were correlated with Leishmania DNA as quantified by real-time PCR. Our results indicate that kinetoplast and nuclear parasite DNA degradation occurs very rapidly after amastigote death. These data add further weight to the argument that PCR assays represent not only a robust method for diagnosis but can also be reliable for monitoring parasite size reduction rate post any intervention (Leishmania-targeting molecules, immunomodulators…). |
| Databáze: | OpenAIRE |
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