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RDT blood spots provide a source of parasite DNA for molecular analysis. In this study, the utility of RDTs for molecular analysis and the performance of different methods for whole genome amplification were investigated.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Methods<\/jats:title>\n                    <jats:p>\n                      Positive\n                      <jats:italic>P. falciparum<\/jats:italic>\n                      RDTs were collected from Kayin, Myanmar from August 2014 to January 2016.\u00a0The RDT samples were stored for 6\u00a0months, 9\u00a0months, 20\u00a0months, 21\u00a0months, and 32\u00a0months before DNA extraction and subsequent molecular analysis of\n                      <jats:italic>P. falciparum kelch 13<\/jats:italic>\n                      (\n                      <jats:italic>pfkelch13<\/jats:italic>\n                      ) mutations,\n                      <jats:italic>P. falciparum multidrug resistance 1<\/jats:italic>\n                      (\n                      <jats:italic>pfmdr1)<\/jats:italic>\n                      , and\n                      <jats:italic>P. falciparum plasmepsin 2<\/jats:italic>\n                      (\n                      <jats:italic>pfplasmepsin2<\/jats:italic>\n                      ) gene amplification. In addition, performance of four whole genome amplification (WGA) kits were compared, including REPLI-g\n                      <jats:sup>\u00ae<\/jats:sup>\n                      , MALBAC\n                      <jats:sup>TM<\/jats:sup>\n                      , PicoPLEX\n                      <jats:sup>\u00ae<\/jats:sup>\n                      , and GenomePlex\n                      <jats:sup>\u00ae<\/jats:sup>\n                      , for which DNA quantity and quality were compared between original DNA and post-WGA products.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Results<\/jats:title>\n                    <jats:p>\n                      The proportion of successful amplification of the different molecular markers was similar between blood spots analysed from RDTs stored for 6, 9, 20, 21, or 32\u00a0months. Successful amplification was dependent on the molecular markers fragment length (\n                      <jats:italic>p value\u2009<\/jats:italic>\n                      &lt;\u20090.05): 18% for a 1245\u00a0bp fragment of\n                      <jats:italic>pfkelch13<\/jats:italic>\n                      , 71% for 364\u00a0bp of\n                      <jats:italic>pfkelch13<\/jats:italic>\n                      , 81% for 87\u00a0bp of\n                      <jats:italic>pfmdr1<\/jats:italic>\n                      , 81% for 108\u00a0bp of\n                      <jats:italic>pfplasmepsin2<\/jats:italic>\n                      . Comparison of the four WGA assay kits showed that REPLI-g\n                      <jats:sup>\u00ae<\/jats:sup>\n                      , MALBAC\n                      <jats:sup>TM<\/jats:sup>\n                      , and PicoPLEX\n                      <jats:sup>\u00ae<\/jats:sup>\n                      increased the quantity of DNA 60 to 750-fold, whereas the ratio of parasite DNA amplification over human DNA was most favourable for MALBAC\n                      <jats:sup>\u00ae<\/jats:sup>\n                      . Sequencing results of\n                      <jats:italic>pfkelch13<\/jats:italic>\n                      ,\n                      <jats:italic>P. falciparum chloroquine resistance transporter<\/jats:italic>\n                      (\n                      <jats:italic>pfcrt<\/jats:italic>\n                      ),\n                      <jats:italic>P. falciparum dihydrofolate reductase<\/jats:italic>\n                      (\n                      <jats:italic>pfdhfr<\/jats:italic>\n                      ) and six microsatellite markers assessed from the post-WGA product was the same as from the original DNA.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Conclusions<\/jats:title>\n                    <jats:p>\n                      Blood spots from RDTs are a good source for molecular analysis of\n                      <jats:italic>P. falciparum<\/jats:italic>\n                      , even after storage up to 32\u00a0months. WGA of RDT-derived parasite DNA reliably increase DNA quantity with sufficient quality for molecular analysis of resistance markers.\n                    <\/jats:p>\n                  <\/jats:sec>","DOI":"10.1186\/s12936-020-03259-9","type":"journal-article","created":{"date-parts":[[2020,5,27]],"date-time":"2020-05-27T07:03:40Z","timestamp":1590563020000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":15,"title":["Utility of Plasmodium falciparum DNA from rapid diagnostic test kits for molecular analysis and whole genome amplification"],"prefix":"10.1186","volume":"19","author":[{"given":"Suttipat","family":"Srisutham","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kanokon","family":"Suwannasin","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Vivek Bhakta","family":"Mathema","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kanlaya","family":"Sriprawat","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Frank M.","family":"Smithuis","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Francois","family":"Nosten","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Nicholas J.","family":"White","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Arjen M.","family":"Dondorp","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0857-1855","authenticated-orcid":false,"given":"Mallika","family":"Imwong","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2020,5,27]]},"reference":[{"key":"3259_CR1","doi-asserted-by":"publisher","first-page":"1491","DOI":"10.1099\/jmm.0.052506-0","volume":"62","author":"JC Mouatcho","year":"2013","unstructured":"Mouatcho JC, Goldring JP. 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