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In Ujjain, central India, the frequency of mutations associated with increased drug tolerance, but not overt resistance to sulfadoxine and pyrimethamine were 9% and\u2009&gt;\u200980%, respectively, in 2009 and 2010, just prior to the introduction of ASP. The frequency of drug resistance associated mutations in Ujjain in 2015\u20132016 after 3\u20134\u00a0years of ASP use, are reported.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Methods<\/jats:title>\n                    <jats:p>\n                      Blood samples from patients with\n                      <jats:italic>P. falciparum<\/jats:italic>\n                      mono-infection verified by microscopy were collected on filter-paper at all nine major pathology laboratories in Ujjain city. Codons\n                      <jats:italic>pfdhfr<\/jats:italic>\n                      16\u2013185,\n                      <jats:italic>pfdhps<\/jats:italic>\n                      436\u2013632 and K13 407\u2013689 were identified by sequencing.\n                      <jats:italic>Pfcrt<\/jats:italic>\n                      K76T and\n                      <jats:italic>pfmdr1<\/jats:italic>\n                      N86Y were identified by restriction fragment length polymorphism.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Results<\/jats:title>\n                    <jats:p>\n                      Sulfadoxine\u2013pyrimethamine resistance-associated\n                      <jats:italic>pfdhfr<\/jats:italic>\n                      108\u00a0N and 59R alleles were found in 100\/104 (96%) and 87\/91 (96%) samples, respectively.\n                      <jats:italic>Pfdhps<\/jats:italic>\n                      437G was found in 10\/105 (10%) samples. Double mutant\n                      <jats:italic>pfdhfr<\/jats:italic>\n                      59R\u2009+\u2009108\u00a0N were found in 75\/81 (93%) samples. Triple mutant\n                      <jats:italic>pfdhfr<\/jats:italic>\n                      59R\u2009+\u2009108\u00a0N and\n                      <jats:italic>pfdhps<\/jats:italic>\n                      437G were found in 6\/78 (8%) samples. Chloroquine-resistance-associated\n                      <jats:italic>pfcrt<\/jats:italic>\n                      76T was found in 102\/102 (100%).\n                      <jats:italic>Pfmdr1<\/jats:italic>\n                      N86 and 86Y were identified in 83\/115 (72%) and 32\/115 (28%) samples, respectively.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Conclusion<\/jats:title>\n                    <jats:p>\n                      The frequency of\n                      <jats:italic>P. falciparum<\/jats:italic>\n                      with reduced susceptibility to sulfadoxine\u2013pyrimethamine remained high, but did not appear to have increased significantly since the introduction of ASP. No polymorphisms in K13 associated with decreased artemisinin susceptibility were found. ASP probably remained effective, supporting continued ASP use.\n                    <\/jats:p>\n                  <\/jats:sec>","DOI":"10.1186\/s12936-020-03274-w","type":"journal-article","created":{"date-parts":[[2020,8,14]],"date-time":"2020-08-14T09:03:00Z","timestamp":1597395780000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":10,"title":["Stable high frequencies of sulfadoxine\u2013pyrimethamine resistance associated mutations and absence of K13 mutations in Plasmodium falciparum 3 and 4\u00a0years after the introduction of artesunate plus sulfadoxine\u2013pyrimethamine in Ujjain, Madhya Pradesh, India"],"prefix":"10.1186","volume":"19","author":[{"given":"Ashish","family":"Pathak","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Andreas","family":"M\u00e5rtensson","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Sudhir","family":"Gawariker","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ashish","family":"Sharma","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Vishal","family":"Diwan","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Manju","family":"Purohit","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5508-9327","authenticated-orcid":false,"given":"Johan","family":"Ursing","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"297","published-online":{"date-parts":[[2020,8,14]]},"reference":[{"key":"3274_CR1","volume-title":"World Malaria Report Country Profiles","author":"WHO","year":"2019","unstructured":"WHO. 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