comparative analysis of multiplex/semi-nested pcr and microscopy for accurate human malaria species diagnosis

Iran is currently striving to eliminate malaria, making it essential to improve the accurate and rapid diagnosis of suspected cases. objectives: this study aimed to evaluate the effectiveness of the multiplex/semi-nested polymerase chain reaction (pcr) method, using specific primers, for the precise diagnosis of human malaria species. methods: seventy-two blood samples from patients and suspected malaria cases were selected and stored at -80°c in the national malaria laboratory. dna extraction was performed to obtain the genetic material for further analysis. multiplex/seminested pcr was conducted, and the results were compared with microscopic examination. results: out of 72 samples, 36 were positive by microscopic analysis, which included: 16 plasmodium falciparum, 16 p. vivax, 1 p. ovale, 1 p. malariae, 1 mixed p. falciparum - p. vivax, and 1 mixed p. falciparum - p. malariae. thirty-three cases were diagnosed through molecular analysis: 16 p. falciparum, 13 p. vivax, 1 p. ovale, 1 p. malariae, and 2 mixed p. falciparum - p. vivax. conclusions: issues, such as false positives, underreporting of mixed infections, and mismatched species identified by microscopic methods need to be addressed and improved to ensure accurate diagnoses.