Candida species are major opportunistic fungal causative agents for significant infections. While C. albicans is considered the most widespread and virulent species, the clinical importance of non-albicans Candida species has substantially risen. This study investigates Palestinian clinical isolates of Candida species by identifying their species type, genotype, mating type, and biofilm formation capability, as well as detecting virulence factors using polymerase chain reaction (PCR). One hundred clinical Candida isolates were obtained from four medical governmental hospitals in Central and Northern Palestine. The PCR technique was used to identify the Candida species and the ABC genotype for the 25S rDNA. The mating type was also determined, besides the detection of hyphal wall protein 1, agglutinin-like sequence genes, phospholipases, and secreted aspartic proteases. Genetic heterogeneity among C. albicans strains was determined by RAPD-PCR typing. C. albicans (53%) and C. tropicalis (15%) are highly represented species. Genotype A was the most prevalent among C. albicans isolates (27%). Notably, the Agglutinin-like sequence genes are present in 59% of the isolates. In contrast, 30% of the isolates have either the SAP9 or/and SAP10 genes. Therefore, the results of RAPD-PCR typing revealed that 14 fragments were produced using the OPI 06 primer with a 100% polymorphism. The 24 C. albicans isolates were clustered into four groups on a 50% cut-off similarity. C. tropicalis was the most frequently isolated non-albicans Candida species. Biofilm formation and the presence of SAP9 and/or SAP10 genes were mainly observed in C. albicans strains, which exhibited low genetic variability.