title: A novel real-time PCR assay for the detection of Plasmodium falciparum and Plasmodium vivax malaria in low parasitized individuals.
abstract: The rapid, accurate diagnosis of Plasmodium spp. is essential for the effective control of malaria, especially in asymptomatic infections. In this study, we developed a sensitive, genus-specific, real-time quantitative PCR assay. It was compared with the microscopic examination of Giemsa-stained blood smears and two different molecular diagnostic techniques: nested PCR and multiplex PCR. For the effective quantitative detection of malaria parasites, all reagents were designed with a lyophilized format in one tube. Plasmodium was detected successfully in all 112 clinically suspected malaria patients, including 32 individuals with low parasitemia (1-100 parasites/μl). The sensitivity threshold was 0.2 parasites/μl and no PCR-positive reaction occurred when malaria parasites were not present. This may be a useful method for detecting malaria parasites in endemic areas.
authors.affiliation: Department of Parasitology, Inje University College of Medicine, Busan 614-735, South Korea.
authors.forename: Seung-Young
authors.initials: SY
authors.lastname: Hwang
authors.forename: So-Hee
authors.initials: SH
authors.lastname: Kim
authors.forename: Ga-Young
authors.initials: GY
authors.lastname: Lee
authors.forename: Vu Thi Thu
authors.initials: VT
authors.lastname: Hang
authors.forename: Chi-Sook
authors.initials: CS
authors.lastname: Moon
authors.forename: Jeong Hwan
authors.initials: JH
authors.lastname: Shin
authors.forename: Wan-Lim
authors.initials: WL
authors.lastname: Koo
authors.forename: Seong-Youl
authors.initials: SY
authors.lastname: Kim
authors.forename: Hae-Joon
authors.initials: HJ
authors.lastname: Park
authors.forename: Han-Oh
authors.initials: HO
authors.lastname: Park
authors.forename: Weon-Gyu
authors.initials: WG
authors.lastname: Kho
mesh_terms: D000818:Animals; D019411:Clinical Laboratory Techniques; D016054:DNA, Protozoan / Q000097:blood / Q000302:isolation & purification*; D006801:Humans; D008288:Malaria / Q000097:blood / Q000175:diagnosis*; D008853:Microscopy / Q000379:methods; D060885:Multiplex Polymerase Chain Reaction / Q000379:methods; D018512:Parasitemia / Q000175:diagnosis*; D010963:Plasmodium falciparum / Q000302:isolation & purification*; D010966:Plasmodium vivax / Q000302:isolation & purification*; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D013045:Species Specificity
chemical_list: D016054:DNA, Protozoan