title: Minority-variant pfcrt K76T mutations and chloroquine resistance, Malawi.
abstract: Genotyping of the chloroquine-resistance biomarker pfcrt (Plasmodium falciparum chloroquine resistance transporter gene) suggests that, in the absence of chloroquine pressure, Plasmodium falciparum parasites in Malawi have reverted to chloroquine sensitivity. However, malaria infections in Africa are commonly polyclonal, and standard PCRs cannot detect minority genotypes if present in <20% of the parasites in an individual host. We have developed a multiple site-specific heteroduplex tracking assay (MSS-HTA) that can detect pfcrt 76T mutant parasites consisting of as little as 1% of the parasite population. In clinical samples, no pfcrt 76T was detected in 87 pregnant Malawian women by standard PCR. However, 22 (25%) contained minority-variant resistant genotypes detected by the MSS-HTA. These results were confirmed by subcloning and sequencing. This finding suggests that the chloroquine-resistant genotype remains common in Malawians and that PCR-undetectable drug-resistant genotypes may be present in disease-endemic populations. Surveillance for minority-variant drug resistant mutations may be useful in making antimalarial drug policy.
authors.affiliation: Department of Epidemiology, University of North Carolina, Chapel Hill, North Carolina 27599-7435, USA.
authors.forename: Jonathan J
authors.initials: JJ
authors.lastname: Juliano
authors.forename: Jesse J
authors.initials: JJ
authors.lastname: Kwiek
authors.forename: Kathryn
authors.initials: K
authors.lastname: Cappell
authors.forename: Victor
authors.initials: V
authors.lastname: Mwapasa
authors.forename: Steven R
authors.initials: SR
authors.lastname: Meshnick
mesh_terms: D000293:Adolescent; D000328:Adult; D000818:Animals; D000962:Antimalarials / Q000494:pharmacology*; D002738:Chloroquine / Q000494:pharmacology*; D004351:Drug Resistance / Q000235:genetics*; D005260:Female; D005838:Genotype; D020180:Heteroduplex Analysis; D006801:Humans; D016778:Malaria, Falciparum / Q000188:drug therapy / Q000453:epidemiology* / Q000235:genetics; D008295:Malawi / Q000453:epidemiology; D026901:Membrane Transport Proteins / Q000235:genetics*; D009692:Nucleic Acid Heteroduplexes / Q000032:analysis / Q000235:genetics*; D010963:Plasmodium falciparum / Q000187:drug effects / Q000235:genetics*; D017354:Point Mutation / Q000235:genetics; D016133:Polymerase Chain Reaction; D011247:Pregnancy; D015995:Prevalence; D015800:Protozoan Proteins / Q000235:genetics*; D012680:Sensitivity and Specificity
chemical_list: D000962:Antimalarials; D026901:Membrane Transport Proteins; D009692:Nucleic Acid Heteroduplexes; C416369:PfCRT protein, Plasmodium falciparum; D015800:Protozoan Proteins; D002738:Chloroquine