[Progress towards the use of DNA microarray technology for the study of wild Plasmodium strains].
Plasmodium falciparum is still a major cause of mortality in the world. Due to growing drug resistance in most endemic countries, the use of tools allowing large-scale analysis of P. falciparum biology is increasingly urgent. In addition to gene sequence data, post-genomic methods including microarray-based transcript profiling allow complete Plasmodium gene expression. However, application of this technology has been limited to study of samples presenting large quantities of total RNA (8 microg to 50 microg). Indeed at least two replicas (one technical and one biological) are necessary to ensure the statistical strength of results. This constraint excludes the use of biological materials hardly available and of wild strain samples. Many methods have been developed to facilitate the use of microarray technology with smaller quantities of total RNA. Currently the use of amplification techniques, various fluorochrome markers, and labelled cDNA and/or RNA probes avoiding all amplification steps, to enhance detection sensitivity has enabled reliable assays to be performed with less material. However these enhancement techniques appear to have a biasing effect on results and the use of powerful new markers is still limited for technical and practical reasons. At the present time radioactive labeling is the most reliable technique for assays using small quantities of total RNA. This approach is not only compatible with competitive hybridization but also enables all microarray screening criteria. Findings from our laboratory support the effectiveness of radioactive labeling for microarray-based determinations on P. falciparum.
Refour
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P
Unité INSERM U511, Immuno-Biologie Cellulaire et Moleculaire des Infections Parasitaires, CHU Pitié-salpêtrière, Université Pierre et Marie Curie Paris VI, Paris, France. refour@ext.jussieu.fr
Gissot
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M
Siau
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A
Mazier
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D
Vaquero
C
C
D000818:Animals; D020411:Oligonucleotide Array Sequence Analysis; D010961:Plasmodium / Q000145:classification* / Q000235:genetics*; D012333:RNA, Messenger / Q000032:analysis
D012333:RNA, Messenger