Parasites, Hosts and Diseases

The Korea Society for Parasitology and Tropical Medicine (대한기생충학·열대의학회)
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Effective High-Throughput Blood Pooling Strategy before DNA Extraction for Detection of Malaria in Low-Transmission Settings

  • Nyunt, Myat Htut (Department of Medical Environmental Biology and Tropical Medicine, School of Medicine, Kangwon National University) ;
  • Kyaw, Myat Phone (Department of Medical Research) ;
  • Thant, Kyaw Zin (Department of Medical Research) ;
  • Shein, Thinzer (Department of Medical Research) ;
  • Han, Soe Soe (Department of Medical Research) ;
  • Zaw, Ni Ni (Department of Medical Research) ;
  • Han, Jin-Hee (Department of Medical Environmental Biology and Tropical Medicine, School of Medicine, Kangwon National University) ;
  • Lee, Seong-Kyun (Department of Medical Environmental Biology and Tropical Medicine, School of Medicine, Kangwon National University) ;
  • Muh, Fauzi (Department of Medical Environmental Biology and Tropical Medicine, School of Medicine, Kangwon National University) ;
  • Kim, Jung-Yeon (Division of Malaria and Parasitic Diseases, National Institute of Health, Centers for Disease Control and Prevention) ;
  • Cho, Shin-Hyeong (Division of Malaria and Parasitic Diseases, National Institute of Health, Centers for Disease Control and Prevention) ;
  • Lee, Sang-Eun (Division of Malaria and Parasitic Diseases, National Institute of Health, Centers for Disease Control and Prevention) ;
  • Yang, Eun-Jeong (Division of Malaria and Parasitic Diseases, National Institute of Health, Centers for Disease Control and Prevention) ;
  • Chang, Chulhun L. (Department of Laboratory Medicine, Pusan National University Yangsan Hospital) ;
  • Han, Eun-Taek (Department of Medical Environmental Biology and Tropical Medicine, School of Medicine, Kangwon National University)
  • Received : 2016.04.18
  • Accepted : 2016.05.27
  • Published : 2016.06.30
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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, $20{\mu}l$ in 1 sample, was optimal, and the parasite density as low as $2p/{\mu}l$ 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.

Keywords

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