en
Scientific article
English

Gene expression profiling of Leishmania (Leishmania) donovani: overcoming technical variation and exploiting biological variation

Published inParasitology, vol. 135, no. 2, p. 183-194
Publication date2008
Abstract

Gene expression profiling is increasingly used in the field of infectious diseases for characterization of host, pathogen and the nature of their interaction. The purpose of this study was to develop a robust, standardized method for comparative expression profiling and molecular characterization of Leishmania donovani clinical isolates. The limitations and possibilities associated with expression profiling in intracellular amastigotes and promastigotes were assessed through a series of comparative experiments in which technical and biological parameters were scrutinized. On a technical level, our results show that it is essential to use parasite harvesting procedures that involve minimal disturbance of the parasite's environment in order to 'freeze' gene expression levels instantly; this is particularly a delicate task for intracellular amastigotes and for specific 'sensory' genes. On the biological level, we demonstrate that gene expression levels fluctuate during in vitro development of both intracellular amastigotes and promastigotes. We chose to use expression-curves rather than single, specific, time-point measurements to capture this biological variation. Intracellular amastigote protocols need further refinement, but we describe a first generation tool for high-throughput comparative molecular characterization of patients' isolates, based on the changing expression profiles of promastigotes during in vitro differentiation.

Keywords
  • Animals
  • DNA Primers/chemistry
  • Gene Expression Profiling/methods
  • Gene Expression Regulation, Developmental
  • Genes, Protozoan/physiology
  • Humans
  • Leishmania donovani/genetics/growth & development/physiology
  • Leishmaniasis, Visceral/parasitology
  • Life Cycle Stages/physiology
  • Polymerase Chain Reaction/methods
  • Time Factors
Citation (ISO format)
DECUYPERE, S. et al. Gene expression profiling of Leishmania (Leishmania) donovani: overcoming technical variation and exploiting biological variation. In: Parasitology, 2008, vol. 135, n° 2, p. 183–194. doi: 10.1017/S0031182007003782
Main files (1)
Article (Accepted version)
accessLevelRestricted
Identifiers
ISSN of the journal0031-1820
487views
0downloads

Technical informations

Creation06/04/2009 9:54:00 AM
First validation06/04/2009 9:54:00 AM
Update time03/14/2023 3:06:11 PM
Status update03/14/2023 3:06:11 PM
Last indexation01/15/2024 6:31:42 PM
All rights reserved by Archive ouverte UNIGE and the University of GenevaunigeBlack