title: A reliable and rapid method for molecular detection of malarial parasites using microwave irradiation and loop mediated isothermal amplification.
abstract: BACKGROUND
  Improved living conditions together with appropriate diagnosis can reduce avoidable malarial deaths substantially. Microscopy remains the gold standard in the diagnosis of malaria. However, rapid molecular diagnostic tests (RmDT) are becoming increasingly important and will, most likely, be the diagnostic techniques of choice in the next years.
  
  
  METHODS
  In this study, a rapid and reliable nucleic acid extraction procedure from human blood and malarial parasites using microwave irradiation as a promising platform is described. In addition, a tailored loop mediated isothermal amplification (LAMP) methodology that utilizes hydroxynaphthol blue (HNB) and Bst 2.0 DNA polymerases in molecular detection of malarial parasites is described.
  
  
  RESULTS
  Following microwave irradiation for DNA isolation, conventional PCR assays were able to detect up to five malaria parasites/μl. The LAMP methodology described here was capable to detect as low as one Plasmodium falciparum parasite/μl after DNA extraction by microwave irradiation. A turnover time of 45 minutes from nucleic acid extraction to final visual read-out was achieved.
  
  
  CONCLUSIONS
  The described procedure offers a cheap, simple and fast method of molecular detection of malaria parasites. This test can easily be performed in basic laboratories. The methodology has been validated as a proof of concept and has specifically be developed for use at low-resource settings. Such RmDTs may aid health providers to make timely therapeutic interventions in malaria endemic regions.
authors.forename: Julia R
authors.initials: JR
authors.lastname: Port
authors.forename: Christian
authors.initials: C
authors.lastname: Nguetse
authors.forename: Selorme
authors.initials: S
authors.lastname: Adukpo
authors.affiliation: Institute of Tropical Medicine, University of Tübingen, Wilhelmstrasse 27, 72074 Tübingen, Germany. velavan@medizin.uni-tuebingen.de.
authors.forename: Thirumalaisamy P
authors.initials: TP
authors.lastname: Velavan
mesh_terms: D000328:Adult; D003365:Costs and Cost Analysis; D016054:DNA, Protozoan / Q000235:genetics / Q000302:isolation & purification*; D006801:Humans; D016778:Malaria, Falciparum / Q000175:diagnosis* / Q000469:parasitology; D008872:Microwaves; D025202:Molecular Diagnostic Techniques / Q000191:economics / Q000379:methods*; D021141:Nucleic Acid Amplification Techniques / Q000191:economics / Q000379:methods*; D010963:Plasmodium falciparum / Q000235:genetics / Q000302:isolation & purification*; D013048:Specimen Handling / Q000191:economics / Q000379:methods*; D013997:Time Factors
chemical_list: D016054:DNA, Protozoan