Effective High-Throughput Blood Pooling Strategy before DNA Extraction for Detection of Malaria in Low-Transmission Settings |
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| Authors: | Myat Htut Nyunt Myat Phone Kyaw Kyaw Zin Thant Thinzer Shein Soe Soe Han Ni Ni Zaw Jin-Hee Han Seong-Kyun Lee Fauzi Muh Jung-Yeon Kim Shin-Hyeong Cho Sang-Eun Lee Eun-Jeong Yang Chulhun L Chang Eun-Taek Han |
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| Institution: | 1Department of Medical Environmental Biology and Tropical Medicine, School of Medicine, Kangwon National University, Chuncheon 24341, Korea;2Department of Medical Research, Yangon, Republic of the Union of Myanmar;3Division of Malaria and Parasitic Diseases, National Institute of Health, Centers for Disease Control and Prevention, Osong 28159, Korea;4Department of Laboratory Medicine, Pusan National University Yangsan Hospital, Yansan 50612, Korea |
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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 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. |
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| Keywords: | Plasmodium falciparum Plasmodium vivax malaria pooling strategy low-transmission setting nested PCR |
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