title: Simplified Pan-species Real-time PCR-based Detection ofPlasmodiumSpp. in Blood Smear.
abstract: BACKGROUND
  We aimed to quicken and simplify the detection ofPlasmodiumin blood samples by developing and testing a pan-Plasmodiumreal-time PCR for accurate screening of individuals suspected of malaria.
  
  
  METHODS
  A single primer/probe set for pan-speciesPlasmodium-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 withP. falciparumandP. vivaxbut 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 forP. vivaxand two as positive forP. 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.
  
  
  CONCLUSIONS
  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 ofPlasmodiumin large numbers of samples.
authors.affiliation: Center for Research of Endemic Parasites of Iran, Tehran University of Medical Sciences, Tehran, Iran; Department of Medical Parasitology & Mycology, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran.
authors.forename: Gholamreza
authors.initials: G
authors.lastname: Hassanpour
authors.affiliation: Department of Medical Parasitology & Mycology, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran.
authors.forename: Hossein
authors.initials: H
authors.lastname: Mirhendi
authors.affiliation: Center for Research of Endemic Parasites of Iran, Tehran University of Medical Sciences, Tehran, Iran; Department of Medical Parasitology & Mycology, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran.
authors.forename: Mehdi
authors.initials: M
authors.lastname: Mohebali
authors.affiliation: Department of Medical Entomology & Vector Control, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran.
authors.forename: Ahmad
authors.initials: A
authors.lastname: Raeisi
authors.affiliation: Department of Epidemiology and Biostatistics, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran.
authors.forename: Hojjat
authors.initials: H
authors.lastname: Zeraati
authors.affiliation: Center for Research of Endemic Parasites of Iran, Tehran University of Medical Sciences, Tehran, Iran; Department of Medical Parasitology & Mycology, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran.
authors.forename: Hossein
authors.initials: H
authors.lastname: Keshavarz
keywords: Diagnosis; FTA cards; Malaria; Plasmodium; Real-time PCR