The root-knot nematode Meloidogyne incognita is a polyphagous pest distributed from temperate to tropical regions. However, the lack of suitable markers leads to a poor knowledge of its population genetic structure and colonization process. Here we describe the first characterization of 15 microsatellite loci from this nematode, that were developed from an enriched genomic library. Although the variability of these microsatellites was generally low, three of them exhibited a significant level of intrapopulation polymorphism, with three to seven alleles detected. The observed and expected heterozygosities ranged from 0.025 to 0.385 and from 0.024 to 0.779, respectively. Thus, these new microsatellite markers have potential value for the implementation of genotyping experiments in this nematode. Furthermore, successful cross-amplification of the variable microsatellite loci in seven other Meloidogyne species provides the opportunity of using these markers for population genetic studies in these damaging plant-parasitic nematodes.