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KMID : 0425120160540030253
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Parasites, Hosts and Diseases
2016 Volume.54 No. 3 p.253 ~ p.259
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Effective High-Throughput Blood Pooling Strategy before DNA Extraction for Detection of Malaria in Low-Transmission Settings
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Myat Htut Nyunt
Myat Phone Kyaw
Kyaw Zin Thant
Thinzer Shein
Han Soe-Soe
Ni Ni Zaw
Kim Jung-Yeon
Cho Shin-Hyeong
Lee Sang-Eun
Yang Eun-Jeong
Han Eun-Taek
Chulhun L. Chang
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Abstract
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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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KEYWORD
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Plasmodium falciparum, Plasmodium vivax, malaria, pooling strategy, low-transmission setting, nested PCR
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