Laila R A Barbosa1,2, Emanuelle L da Silva1,3, Anne C G de Almeida1,2,4, Yanka E A R Salazar1, André M Siqueira5, Maria das Graças Costa Alecrim1,2, José Luiz Fernandes Vieira6, Quique Bassat7,8,9,10,11, Marcus V G de Lacerda1,2,12, Wuelton M Monteiro1,2, Gisely C Melo1,2. 1. Fundação de Medicina Tropical Dr. Heitor Vieira Dourado, Instituto de Pesquisa Clínica Carlos Borborema, Manaus 69040-200, AM, Brazil. 2. Programa de Pós-Graduação em Medicina Tropical, Universidade do Estado do Amazonas, Manaus 69040-200, AM, Brazil. 3. UNINILTONLINS-Universidade Nilton Lins, Unicenter, Manaus 69058-030, AM, Brazil. 4. FAMETRO-Faculdade Metropolitana de Manaus, Campus Central, Av. Constantino Nery, Chapada, Manaus 69050-000, AM, Brazil. 5. Fiocruz-Manguinhos-Fundação Oswaldo Cruz, Instituto de Pesquisa Clínica Evandro Chagas, Rio de Janeiro 21040-900, RJ, Brazil. 6. Instituto de Ciências da Saúde, Universidade Federal do Pará, Belém 66010-010, PA, Brazil. 7. ISGlobal, Hospital Clínic, Universitat de Barcelona, 08036 Barcelona, Spain. 8. Centro de Investigação em Saúde de Manhiça (CISM), Fundação Clínic per la Recerca Biomédica, Maputo 1929, Mozambique. 9. Catalan Institution for Research and Advanced Studies (ICREA), Campus Clínic, Pg. Lluís Companys 23, 08010 Barcelona, Spain. 10. Pediatric Infectious Diseases Unit, Pediatrics Department, Hospital Sant Joan de Déu, University of Barcelona, 1867 Barcelona, Spain. 11. Consorcio de Investigación Biomédica en Red de Epidemiología y Salud Pública (CIBERESP), 28029 Madrid, Spain. 12. Instituto Leônidas & Maria Deane, ILMD-Fiocruz-Fundação Oswaldo Cruz, Manaus 69057-070, AM, Brazil.
Abstract
BACKGROUND: Early recurrence of Plasmodium vivax is a challenge for malaria control in the field, particularly because this species is associated with lower parasitemia, which hinders diagnosis and monitoring through blood smear testing. Early recurrences, defined as the persistence of parasites in the peripheral blood despite adequate drug dosages, may arise from resistance to chloroquine. The objective of the study was to estimate early recurrence of P. vivax in the Brazilian Amazon by using a highly-sensitive detection method, in this case, PCR. METHODS: An ultra-sensitive qPCR that targeted mitochondrial DNA was used to compare a standard qPCR that targeted 18S rDNA to detect early recurrence of P. vivax in very low densities in samples from patients treated with chloroquine. RESULTS: Out of a total of 312 cases, 29 samples (9.3%) were characterized as recurrences, from which 3.2% (10/312) were only detected through ultra-sensitive qPCR testing. CONCLUSIONS: Studies that report the detection of P. vivax early recurrences using light microscopy may severely underestimate their true incidence.
BACKGROUND: Early recurrence of Plasmodium vivax is a challenge for malaria control in the field, particularly because this species is associated with lower parasitemia, which hinders diagnosis and monitoring through blood smear testing. Early recurrences, defined as the persistence of parasites in the peripheral blood despite adequate drug dosages, may arise from resistance to chloroquine. The objective of the study was to estimate early recurrence of P. vivax in the Brazilian Amazon by using a highly-sensitive detection method, in this case, PCR. METHODS: An ultra-sensitive qPCR that targeted mitochondrial DNA was used to compare a standard qPCR that targeted 18S rDNA to detect early recurrence of P. vivax in very low densities in samples from patients treated with chloroquine. RESULTS: Out of a total of 312 cases, 29 samples (9.3%) were characterized as recurrences, from which 3.2% (10/312) were only detected through ultra-sensitive qPCR testing. CONCLUSIONS: Studies that report the detection of P. vivax early recurrences using light microscopy may severely underestimate their true incidence.
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