Tehran University of Medical Sciences Iranian Journal of Parasitology 1735-7020 11 4 2016 12 25 463 470 Gholamreza HASSANPOUR Center for Research of Endemic Parasites of Iran, Tehran University of Medical Sciences, Tehran, Iran AND Department of Medical Parasitology & Mycology, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran Hossein MIRHENDI Department of Medical Parasitology & Mycology, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran Mehdi MOHEBALI Center for Research of Endemic Parasites of Iran, Tehran University of Medical Sciences, Tehran, Iran AND Department of Medical Parasitology & Mycology, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran Ahmad RAEISI Department of Medical Entomology & Vector Control, School of Public Health, Tehran University of Medical Sci-ences, Tehran, Iran Hojjat ZERAATI Department of Epidemiology and Biostatistics, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran Hossein KESHAVARZ Center for Research of Endemic Parasites of Iran, Tehran University of Medical Sciences, Tehran, Iran AND Department of Medical Parasitology & Mycology, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran 2016 12 27 2016 12 27 Background: We aimed to quicken and simplify the detection of Plasmodium in blood samples by developing and testing a pan-Plasmodium real-time PCR for accurate screening of individuals suspected of malaria. Methods: A single primer/probe set for pan-species Plasmodium-specific real time PCR targeting a conserved region of the small subunit 18S ribosomal DNA was designed and evaluated for rapid diagnosis and screening of malaria infections using dried blood smears. FTA cards were used for rapid and simple DNA extraction. Results: The primers and probes showed a positive response with the DNA extracted from bloods infected with P. falciparum and P. vivax but not with DNA extracted from various smears from uninfected blood samples. Seven positive cases positive by both microscopy and nested PCR were found among 280 blood samples taken from in South and Southeast Iran. Five samples were identified as positive for P. vivax and two as positive for P. falciparum. All positive samples were positive by real-time PCR. Furthermore, all 38-blood samples positive by microscopy were positive by real-time PCR. No microscopy-negative samples were positive by real-time PCR. Conclusion: By using a simple FTA card for DNA extraction and by application of the real-time PCR developed in this study, sensitivity similar to nested-PCR and microscopy was achieved. This format simplifies the detection of Plasmodium in large numbers of samples. https://ijpa.tums.ac.ir/index.php/ijpa/article/view/1364 https://ijpa.tums.ac.ir/index.php/ijpa/article/download/1364/640