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There have been stories of increasing numbers of circumstances of malaria among migrants and travelers. Although microscopic examination of blood smears stays the "gold normal" in prognosis, this method suffers from insufficient sensitivity and requires considerable expertise. To improve analysis, a multiplex actual-time PCR was developed. One set of generic primers focusing on a extremely conserved area of the 18S rRNA gene of the genus Plasmodium was designed; the primer set was polymorphic enough internally to design four species-particular probes for P. falciparum, P. vivax, P. malarie, and P. ovale. Real-time PCR with species-particular probes detected one plasmid copy of P. falciparum, P. vivax, P. malariae, and P. ovale particularly. The identical sensitivity was achieved for all species with real-time PCR with the 18S screening probe. Ninety-seven blood samples have been investigated. For sixty six of them (60 patients), microscopy and real-time PCR outcomes have been compared and had a crude settlement of 86% for the detection of plasmodia. Discordant outcomes were reevaluated with clinical, molecular, and sequencing information to resolve them. All 9 discordances between 18S screening PCR and microscopy have been resolved in favor of the molecular method, as had been eight of nine discordances on the species level for the species-specific PCR among the 31 samples positive by both methods. The opposite 31 blood samples had been examined to monitor the antimalaria remedy in seven patients. The number of parasites measured by actual-time PCR fell rapidly for six out of seven patients in parallel to parasitemia decided microscopically. This suggests a job of quantitative PCR for the monitoring of patients receiving antimalaria therapy.
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