Document Detail


Real-time polymerase chain reaction assay for the rapid detection and characterization of chloroquine-resistant Plasmodium falciparum malaria in returned travelers.
MedLine Citation:
PMID:  16447106     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
BACKGROUND: Imported drug-resistant malaria is a growing problem in industrialized countries. Rapid and accurate diagnosis is essential to prevent malaria-associated mortality in returned travelers. However, outside of a limited number of specialized centers, the microscopic diagnosis of malaria is slow, unreliable, and provides little information about drug resistance. Molecular diagnostics have the potential to overcome these limitations. OBJECTIVE: We developed and evaluated a rapid, real-time polymerase chain reaction (PCR) assay to detect Plasmodium falciparum malaria and chloroquine (CQ)-resistance determinants in returned travelers who are febrile. METHODS: A real-time PCR assay based on detection of the K76T mutation in PfCRT (K76T) of P. falciparum was developed on a LightCycler platform (Roche). The performance characteristics of the real-time assay were compared with those of the nested PCR-restriction fragment-length polymorphism (RFLP) and the sequence analyses of samples obtained from 200 febrile returned travelers, who included 125 infected with P. falciparum (48 of whom were infected CQ-susceptible [K76] and 77 of whom were CQ-resistant [T76] P. falciparum), 22 infected with Plasmodium vivax, 10 infected with Plasmodium ovale, 3 infected with Plasmodium malariae malaria, and 40 infected with other febrile syndromes. All patient samples were coded, and all analyses were performed blindly. RESULTS: The real-time PCR assay detected multiple pfcrt haplotypes associated with CQ resistance in geographically diverse malaria isolates acquired by travelers. Compared with nested-PCR RFLP (the reference standard), the real-time assay was 100% sensitive and 96.2% specific for detection of the P. falciparum K76T mutation. CONCLUSION: This assay is rapid, sensitive, and specific for the detection and characterization of CQ-resistant P. falciparum malaria in returned travelers. This assay is automated, standardized, and suitable for routine use in clinical diagnostic laboratories.
Authors:
Gabriella A Farcas; Rainer Soeller; Kathleen Zhong; Alireza Zahirieh; Kevin C Kain
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't     Date:  2006-01-25
Journal Detail:
Title:  Clinical infectious diseases : an official publication of the Infectious Diseases Society of America     Volume:  42     ISSN:  1537-6591     ISO Abbreviation:  Clin. Infect. Dis.     Publication Date:  2006 Mar 
Date Detail:
Created Date:  2006-01-31     Completed Date:  2006-10-16     Revised Date:  2007-11-15    
Medline Journal Info:
Nlm Unique ID:  9203213     Medline TA:  Clin Infect Dis     Country:  United States    
Other Details:
Languages:  eng     Pagination:  622-7     Citation Subset:  IM    
Affiliation:
Tropical Disease Unit, University Health Network-Toronto General Hospital, Toronto, Ontario, Canada.
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MeSH Terms
Descriptor/Qualifier:
Animals
Antimalarials / pharmacology*
Chloroquine / pharmacology*
Drug Resistance*
Humans
India
Malaria, Falciparum / diagnosis*,  drug therapy,  parasitology*
Membrane Proteins / genetics
Membrane Transport Proteins
Mutation
Plasmodium falciparum / genetics
Polymerase Chain Reaction / methods*
Protozoan Proteins
Sensitivity and Specificity
Travel
Chemical
Reg. No./Substance:
0/Antimalarials; 0/Membrane Proteins; 0/Membrane Transport Proteins; 0/PfCRT protein, Plasmodium falciparum; 0/Protozoan Proteins; 54-05-7/Chloroquine
Comments/Corrections
Comment In:
Clin Infect Dis. 2006 Jun 15;42(12):1806-7   [PMID:  16705592 ]

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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