The Andean native potato is an important food security crop and constitutes a gene pool for potato-breeding. To establish the current state of the native potatoes diversity in the influence zone of the Camisea Gas Project, North Ayacucho, Peru, landraces (n = 144) were collected. Three sampling populations (Anco, Chungui and Ticllas) were resolved using 10 simple sequence repeats (SSRs) markers that amplified 67 polymorphic alleles. Principal component, correspondence and cluster analysis revealed a minimum set of six SSRs to achieve DNA fingerprinting and cost-effective genetic diversity analysis. Analysis of molecular variance (AMOVA), average fixation index (FST = 0.279), and genetic distance index (SB) indicated that the genetic diversity of the native potatoes is high at the intra- and inter-population levels, and each of the three sampling populations constituted closed populations. The three populations were genetically distinct and contained unique genotypes and exclusive alleles. Misidentified native potatoes (10.41%) were found.