title: Monitoring chloroquine resistance using Plasmodium falciparum parasites isolated from wild mosquitoes in Tanzania.
abstract: Monitoring antimalarial drug resistance is a useful epidemiologic tool and provides early detection of resistance foci. Using DNA extracted from the head/thorax of wild mosquitoes collected from Bagamoyo Coastal Tanzania, samples infected by Plasmodium falciparum (N = 89, in 2002 and N = 249 in 2004) were screened by nested polymerase chain reaction (PCR)-restriction fragment length polymorphism (RFLP) assay for mutations at Pfcrt76 and Pfmdr1-86 associated with chloroquine (CQ) resistance. The majority of isolates were of single infection (71%), and the prevalence of mutant alleles of Pfcrt76 decreased from 64.5% in 2002 to 16% in 2004; likewise, mutant Pfmdr1-86 alleles decreased from 46.6% to 2.7%. Overall, there was a decline of mutant isolates by a factor of 17 and 4 for Pfmdr1 and Pfcrt, respectively. In contrast, isolates with wild-type alleles increased significantly from < 20% in 2002 to 67.6% for Pfcrt76 and 83.5% for Pfmdr1-86 in 2004. This observation suggest a biologic trend of decrease of CQ mutants and a subsequent increase of CQ susceptible parasites in circulation after the discontinued use of CQ in 2001 as a first-line drug in Tanzania. High prevalence of susceptible P. falciparum found in circulation not only supports other reports of a decline of mutant parasites after a reduction of drug selection pressure but suggests that the fitness cost is high in mutant parasites. Typing parasite isolates from infected mosquitoes, an alternative means of data collection, has the potential to increase the spatial and temporal coverage, and this approach is practical in highly endemic regions of Africa.
authors.affiliation: Department of Vector Ecology and Environment, Institute of Tropical Medicine, Nagasaki University, Nagasaki, Japan. emmatemu@net.nagasaki-u.ca.jp
authors.forename: Emmanuel A
authors.initials: EA
authors.lastname: Temu
authors.forename: Issac
authors.initials: I
authors.lastname: Kimani
authors.forename: Nabuko
authors.initials: N
authors.lastname: Tuno
authors.forename: Hitoshi
authors.initials: H
authors.lastname: Kawada
authors.forename: Japhet N
authors.initials: JN
authors.lastname: Minjas
authors.forename: Masahiro
authors.initials: M
authors.lastname: Takagi
mesh_terms: D000818:Animals; D000962:Antimalarials / Q000494:pharmacology; D002738:Chloroquine / Q000494:pharmacology*; D009033:Culicidae / Q000469:parasitology*; D016903:Drug Monitoring / Q000379:methods; D004351:Drug Resistance; D018432:Drug Resistance, Multiple / Q000235:genetics; D005838:Genotype; D010963:Plasmodium falciparum / Q000187:drug effects* / Q000235:genetics / Q000302:isolation & purification; D013636:Tanzania
chemical_list: D000962:Antimalarials; D002738:Chloroquine