title: Improved assay to detect Plasmodium falciparum using an uninterrupted, semi-nested PCR and quantitative lateral flow analysis.
abstract: BACKGROUND
  A rapid, non-invasive, and inexpensive point-of-care (POC) diagnostic for malaria followed by therapeutic intervention would improve the ability to control infection in endemic areas.
  
  
  METHODS
  A semi-nested PCR amplification protocol is described for quantitative detection of Plasmodium falciparum and is compared to a traditional nested PCR. The approach uses primers that target the P. falciparum dihydrofolate reductase gene.
  
  
  RESULTS
  This study demonstrates that it is possible to perform an uninterrupted, asymmetric, semi-nested PCR assay with reduced assay time to detect P. falciparum without compromising the sensitivity and specificity of the assay using saliva as a testing matrix.
  
  
  CONCLUSIONS
  The development of this PCR allows nucleic acid amplification without the need to transfer amplicon from the first PCR step to a second reaction tube with nested primers, thus reducing both the chance of contamination and the time for analysis to < two hours. Analysis of the PCR amplicon yield was adapted to lateral flow detection using the quantitative up-converting phosphor (UCP) reporter technology. This approach provides a basis for migration of the assay to a POC microfluidic format. In addition the assay was successfully evaluated with oral samples. Oral fluid collection provides a simple non-invasive method to collect clinical samples.
authors: Department of Basic Science, NYU College of Dentistry, New York, NY 10010, USA.
authors: Serge Y
authors: SY
authors: Ongagna-Yhombi
authors: Paul
authors: P
authors: Corstjens
authors: Eran
authors: E
authors: Geva
authors: William R
authors: WR
authors: Abrams
authors: Cheryl A
authors: CA
authors: Barber
authors: Daniel
authors: D
authors: Malamud
authors: Sungano
authors: S
authors: Mharakurwa
mesh_terms: D006801:Humans; D016778:Malaria, Falciparum / Q000175:diagnosis* / Q000469:parasitology*; D059208:Parasite Load; D010963:Plasmodium falciparum / Q000302:isolation & purification*; D016133:Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity