Prevalence of pfcrt mutations in Congolese and Malawian Plasmodium falciparum isolates as determined by a new Taqman assay

A real-time PCR assay was developed to detect the K76T point mutation in the Plasmodium falciparum putative chloroquine resistance transporter gene. The assay was used with malaria positive clinical isolates from Rutshuru in the eastern part of the Democratic Republic of the Congo (DRC) and from Mal...

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Bibliographic Details
Published in:Acta tropica 2005, Vol.93 (1), p.97-106
Main Authors: Wilson, Paul E., Kazadi, Walter, Kamwendo, Deborah Demster, Mwapasa, Victor, Purfield, Anne, Meshnick, Steven R.
Format: Article
Language:English
Subjects:
Animals
Biological and medical sciences
Child, Preschool
Chloroquine
Chloroquine - therapeutic use
Chloroquine resistance transporter
Democratic Republic of the Congo
DNA, Protozoan - genetics
Drug resistance
Drug Resistance - genetics
Female
General aspects
Genotype
Humans
Infant
Malaria, Falciparum - drug therapy
Malaria, Falciparum - parasitology
Malawi
Medical sciences
Membrane Proteins - genetics
Membrane Transport Proteins
Plasmodium falciparum
Plasmodium falciparum - genetics
Plasmodium falciparum - isolation & purification
Point Mutation
Polymerase Chain Reaction
Pregnancy
Protozoan Proteins
Sensitivity and Specificity
Sequence Analysis, DNA
Single nucleotide polymorphism
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Summary:A real-time PCR assay was developed to detect the K76T point mutation in the Plasmodium falciparum putative chloroquine resistance transporter gene. The assay was used with malaria positive clinical isolates from Rutshuru in the eastern part of the Democratic Republic of the Congo (DRC) and from Malawi. The K76T mutation was found in 52/56 (93%) clinical isolates from the DRC, where chloroquine resistance is high, but in none of the 12 isolates tested from Malawi where chloroquine is now rarely used. Sixteen percent of specimens from the DRC had detectable levels of both wild-type and mutant alleles. The real-time PCR results were compared to results from a nested allele-specific PCR assay and from direct DNA sequencing. Using allele-specific PCR as the reference method, the new assay is 100% sensitive and specific towards the mutant allele. In addition to its low per-test cost, the new assay is fast, easily automated, sensitive and well-suited to large-scale epidemiological studies.
ISSN:0001-706X
1873-6254
DOI:10.1016/j.actatropica.2004.09.010