In-vitro maintenance and microscopic characterization of Plasmodium vivax using human reticulocyte culture: implications for diagnostic and drug development

Increasing reports of drug-resistant Plasmodium vivax have intensified the need for efficient In-vitro cultivation systems to support diagnostic and therapeutic research. To establish and evaluate a reproducible In-vitro cultivation protocol for P. vivax using human reticulocytes and assess parasite growth through microscopy and rapid diagnostic testing (RDT). To establish and evaluate a reproducible in vitro cultivation protocol for P. vivax using human reticulocytes and assess parasite growth through microscopy and RDT. Cultured parasites exhibited typical P. vivax stages single chromatin rings, amoeboid trophozoites, and schizonts with an observed parasitaemia increase of 2.5-fold within 72 h. Infected erythrocytes appeared enlarged and deformed, consistent with active infection. pLDH antigen detection confirmed metabolic activity throughout culture cycles. The optimized In-vitro culture method reliably supports successive P. vivax generations, enabling consistent production of native pLDH for diagnostic validation, antimalarial screening, and genomic applications. This approach may facilitate new strategies for combating P. vivax resistance.