Development Of A Plasmodium PCR For Monitoring Efficacy Of Antimalarial Treatment

There have been reviews of accelerating numbers of cases of malaria amongst migrants and travelers. Although microscopic examination of blood smears remains the "gold commonplace" in prognosis, this methodology suffers from insufficient sensitivity and requires considerable experience. To enhance diagnosis, a multiplex real-time PCR was developed. One set of generic primers focusing on a extremely conserved region of the 18S rRNA gene of the genus Plasmodium was designed; the primer set was polymorphic sufficient internally to design 4 species-specific probes for BloodVitals test 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 specifically. 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 66 of them (60 patients), microscopy and real-time PCR outcomes have been in contrast and had a crude agreement of 86% for the detection of plasmodia. Discordant results have been reevaluated with clinical, molecular, and sequencing data to resolve them. All 9 discordances between 18S screening PCR and microscopy had been resolved in favor of the molecular technique, as were eight of 9 discordances at the species degree for the species-particular PCR among the 31 samples positive by each methods. The opposite 31 blood samples have been examined to monitor the antimalaria therapy in seven patients. The variety of parasites measured by actual-time PCR fell quickly for six out of seven patients in parallel to parasitemia determined microscopically. This suggests a job of quantitative PCR for the monitoring of patients receiving antimalaria therapy.



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