title: Effective High-Throughput Blood Pooling Strategy before DNA Extraction for Detection of Malaria in Low-Transmission Settings.
abstract: In the era of (pre) elimination setting, the prevalence of malaria has been decreasing in most of the previously endemic areas. Therefore, effective cost- and time-saving validated pooling strategy is needed for detection of malaria in low transmission settings. In this study, optimal pooling numbers and lowest detection limit were assessed using known density samples prepared systematically, followed by genomic DNA extraction and nested PCR. Pooling strategy that composed of 10 samples in 1 pool, 20 µl in 1 sample, was optimal, and the parasite density as low as 2 p/µl for both falciparum and vivax infection was enough for detection of malaria. This pooling method showed effectiveness for handling of a huge number of samples in low transmission settings (<9% positive rate). The results indicated that pooling of the blood samples before DNA extraction followed by usual nested PCR is useful and effective for detection of malaria in screening of hidden cases in low-transmission settings.
authors.affiliation: Department of Medical Environmental Biology and Tropical Medicine, School of Medicine, Kangwon National University, Chuncheon 24341, Korea.|Department of Medical Research, Yangon, Republic of the Union of Myanmar.
authors.forename: Myat Htut
authors.initials: MH
authors.lastname: Nyunt
authors.affiliation: Department of Medical Research, Yangon, Republic of the Union of Myanmar.
authors.forename: Myat Phone
authors.initials: MP
authors.lastname: Kyaw
authors.affiliation: Department of Medical Research, Yangon, Republic of the Union of Myanmar.
authors.forename: Kyaw Zin
authors.initials: KZ
authors.lastname: Thant
authors.affiliation: Department of Medical Research, Yangon, Republic of the Union of Myanmar.
authors.forename: Thinzer
authors.initials: T
authors.lastname: Shein
authors.affiliation: Department of Medical Research, Yangon, Republic of the Union of Myanmar.
authors.forename: Soe Soe
authors.initials: SS
authors.lastname: Han
authors.affiliation: Department of Medical Research, Yangon, Republic of the Union of Myanmar.
authors.forename: Ni Ni
authors.initials: NN
authors.lastname: Zaw
authors.affiliation: Department of Medical Environmental Biology and Tropical Medicine, School of Medicine, Kangwon National University, Chuncheon 24341, Korea.
authors.forename: Jin-Hee
authors.initials: JH
authors.lastname: Han
authors.affiliation: Department of Medical Environmental Biology and Tropical Medicine, School of Medicine, Kangwon National University, Chuncheon 24341, Korea.
authors.forename: Seong-Kyun
authors.initials: SK
authors.lastname: Lee
authors.affiliation: Department of Medical Environmental Biology and Tropical Medicine, School of Medicine, Kangwon National University, Chuncheon 24341, Korea.
authors.forename: Fauzi
authors.initials: F
authors.lastname: Muh
authors.affiliation: Division of Malaria and Parasitic Diseases, National Institute of Health, Centers for Disease Control and Prevention, Osong 28159, Korea.
authors.forename: Jung-Yeon
authors.initials: JY
authors.lastname: Kim
authors.affiliation: Division of Malaria and Parasitic Diseases, National Institute of Health, Centers for Disease Control and Prevention, Osong 28159, Korea.
authors.forename: Shin-Hyeong
authors.initials: SH
authors.lastname: Cho
authors.affiliation: Division of Malaria and Parasitic Diseases, National Institute of Health, Centers for Disease Control and Prevention, Osong 28159, Korea.
authors.forename: Sang-Eun
authors.initials: SE
authors.lastname: Lee
authors.affiliation: Division of Malaria and Parasitic Diseases, National Institute of Health, Centers for Disease Control and Prevention, Osong 28159, Korea.
authors.forename: Eun-Jeong
authors.initials: EJ
authors.lastname: Yang
authors.affiliation: Department of Laboratory Medicine, Pusan National University Yangsan Hospital, Yansan 50612, Korea.
authors.forename: Chulhun L
authors.initials: CL
authors.lastname: Chang
authors.affiliation: Department of Medical Environmental Biology and Tropical Medicine, School of Medicine, Kangwon National University, Chuncheon 24341, Korea.
authors.forename: Eun-Taek
authors.initials: ET
authors.lastname: Han
mesh_terms: D001769:Blood / Q000469:parasitology*; D016054:DNA, Protozoan / Q000032:analysis* / Q000235:genetics; D006801:Humans; D008288:Malaria / Q000175:diagnosis*; D008403:Mass Screening / Q000379:methods*; D025202:Molecular Diagnostic Techniques / Q000379:methods; D010274:Parasitology / Q000379:methods*; D010961:Plasmodium / Q000235:genetics / Q000302:isolation & purification*; D016133:Polymerase Chain Reaction / Q000379:methods; D013048:Specimen Handling / Q000379:methods*
chemical_list: D016054:DNA, Protozoan
keywords: Plasmodium falciparum; Plasmodium vivax; low-transmission setting; malaria; nested PCR; pooling strategy