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This study assessed the frequency of\n                      <jats:italic toggle=\"yes\">pfcrt<\/jats:italic>\n                      mutations and\n                      <jats:italic toggle=\"yes\">pfplasmepsin2\/3<\/jats:italic>\n                      gene amplifications across Southeast Asia over 25 years, which is critical for tracking resistant parasites. A total of 898\n                      <jats:italic toggle=\"yes\">P<\/jats:italic>\n                      .\n                      <jats:italic toggle=\"yes\">falciparum<\/jats:italic>\n                      isolates collected from Thailand, Myanmar, Cambodia, Laos, and Vietnam between 1995 and 2023 were analyzed for\n                      <jats:italic toggle=\"yes\">pfcrt<\/jats:italic>\n                      mutations,\n                      <jats:italic toggle=\"yes\">pfplasmepsin2<\/jats:italic>\n                      copy number, and microsatellite variation around the\n                      <jats:italic toggle=\"yes\">pfcrt<\/jats:italic>\n                      locus. During the study period,\n                      <jats:italic toggle=\"yes\">pfcrt<\/jats:italic>\n                      mutations in Cambodia across the study period showed that H97Y had the highest prevalence at 15%, followed by T93S at 8%, while I218F was common in 24% of specimens in Srisaket and Ubon in Thailand at 24.44%, followed by G353V at 20%, T93S at 17%, F145I at 13%, and H97Y at 4%. In Tak, Thailand, a mutation was found only in 1995 with 40% at I218F and the remaining wild-type. Yala in Thailand and Kayin State in Myanmar remained wild-type. Vietnam showed frequent T93S at 21%. The prevalence of\n                      <jats:italic toggle=\"yes\">pfcrt<\/jats:italic>\n                      mutations observed in this study changed throughout the study period. Over 65% of parasites with\n                      <jats:italic toggle=\"yes\">pfcrt<\/jats:italic>\n                      mutations at positions 93, 97, 145, 218, 343, and 353 also had\n                      <jats:italic toggle=\"yes\">pfplasmepsin2<\/jats:italic>\n                      amplification. Temporal analysis revealed that in Cambodia,\n                      <jats:italic toggle=\"yes\">pfplasmepsin2<\/jats:italic>\n                      CNV emerged first, peaking at 88% prevalence in 2015 after the introduction of DHA-piperaquine, with the subsequent increase of\n                      <jats:italic toggle=\"yes\">pfcrt<\/jats:italic>\n                      mutations H97Y and G353V. In Eastern Thailand, both\n                      <jats:italic toggle=\"yes\">pfcrt<\/jats:italic>\n                      mutations and\n                      <jats:italic toggle=\"yes\">pfplasmepsin2<\/jats:italic>\n                      CNV were highly prevalent, exceeding 70% during 2015\u20132018. In Vietnam, rapid increases in both markers were observed after 2012, reaching peak by 2017. Microsatellite analysis revealed reduced genetic diversity around mutant\n                      <jats:italic toggle=\"yes\">pfcrt<\/jats:italic>\n                      alleles, indicating selective sweeps. This study demonstrates that frequencies of\n                      <jats:italic toggle=\"yes\">pfcrt<\/jats:italic>\n                      mutations and\n                      <jats:italic toggle=\"yes\">pfplasmepsin2<\/jats:italic>\n                      amplification linked to piperaquine resistance have changed over time, highlighting the importance of ongoing genetic monitoring to inform strategies for preserving artemisinin-based combination therapy efficacy.\n                    <\/jats:p>\n                  <\/jats:sec>","DOI":"10.1128\/aac.01117-25","type":"journal-article","created":{"date-parts":[[2026,2,17]],"date-time":"2026-02-17T14:00:29Z","timestamp":1771336829000},"update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":3,"title":["Changing genetic profiles of\n                    <i>Plasmodium falciparum<\/i>\n                    piperaquine resistance in Southeast Asia over 25 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