Journal
ACTA TROPICA
Volume 93, Issue 1, Pages 97-106Publisher
ELSEVIER SCIENCE BV
DOI: 10.1016/j.actatropica.2004.09.010
Keywords
Plasmodium falciparum; single nucleotide polymorphism; membrane proteins/genetics; genotype; chloroquine; chloroquine; resistance transporter; polymerase chain reaction; drug resistance; Democratic Republic of the Congo; Malawi
Categories
Funding
- NIAID NIH HHS [R01 AI 49084] Funding Source: Medline
- NICHD NIH HHS [U01 HD043475-01] Funding Source: Medline
- PHS HHS [S1935-21/21] Funding Source: Medline
- EUNICE KENNEDY SHRIVER NATIONAL INSTITUTE OF CHILD HEALTH &HUMAN DEVELOPMENT [U01HD043475] Funding Source: NIH RePORTER
- NATIONAL INSTITUTE OF ALLERGY AND INFECTIOUS DISEASES [R01AI049084] Funding Source: NIH RePORTER
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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. (C) 2004 Elsevier B.V. All rights reserved.
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