(1) Background: Malaria is a public health problem worldwide. Despite global efforts to control it, antimalarial drug resistance remains a great challenge. In 2009, our team identified, for the first time in Brazil, chloroquine (CQ)-susceptiblePlasmodium falciparumparasites in isolates from the Brazilian Amazon. The present study extends those observations to include survey samples from 2010 to 2018 from the Amazonas and Acre states for the purpose of trackingpfcrtmolecular changes inP. falciparumparasites. (2) Objective: to investigate SNPs in theP. falciparumgene associated with chemoresistance to CQ (pfcrt).(3) Methods: Sixty-sixP. falciparumsamples from the Amazonas and Acre states were collected from 2010 to 2018 in patients diagnosed at the Reference Research Center for Treatment and Diagnosis of Malaria (CPD-Mal/Fiocruz), FMT-HVD and Acre Health Units. These samples were subjected to PCR and DNA Sanger sequencing to identify mutations inpfcrt(C72S, M74I, N75E, and K76T).(4) Results: Of the 66P. falciparumsamples genotyped forpfcrt, 94% carried CQ-resistant genotypes and only 4 showed a CQpfcrtsensitive-wild type genotype, i.e., 1 from Barcelos and 3 from Manaus. (5) Conclusion: CQ-resistantP. falciparumpopulations are fixed, and thus, CQ cannot be reintroduced in malaria falciparum therapy.