Identification of Acanthamoeba spp. isolates maintained at the amibiasis laboratory of Universidad Central de Venezuela, USING PCR-RFLP

摘要

Acanthamoeba genus includes some species related to pathology in humans. Their identification and classification was made by morphology, but it is a subjective technique and has low sensitivity and specificity. For these reasons, it is important to incorporate molecular tools. The objective of this research was to characterize morphologic and molecular features of 24 Acanthamoeba isolates, maintained at Laboratorio de AmibiasisEscuela of BioanálisisUCV, previously identified as Acanthamoeba , using light microscopy and morphological criteria according to Pussard and Pons, 1977 and PCRRFLP (HinffI, HhaI and HaeIII) respectively. Morphologically, none of the strains belonged to group I. 45.8% of all isolates presented compatible morphology with Group II, other 45.8% with group III and 8.4% with both groups. Molecularly, 50% of the isolates amplified a 900 pb product and the other 50%, a 700 pb product. Isolates of Group III could not be molecularly characterized by PCRRFLP, since the digestion pattern have no coincidence with previous reports. Only 33% of strains were finally identified: A. polyphaga (A9, A12, A13, A14), A. castellanii (A26, A27, A28 and A29) and A. castellanii or A. polyphaga (A15, A25 and A30). The identified species are consistent with the most common pathogens described at the literature.