Mitochondrial DNA targets increase sensitivity of malaria detection using loop-mediated isothermal amplification.
Loop-mediated isothermal amplification (LAMP) of DNA offers the ability to detect very small quantities of pathogen DNA following minimal tissue sample processing and is thus an attractive methodology for point-of-care diagnostics. Previous attempts to diagnose malaria by the use of blood samples and LAMP have targeted the parasite small-subunit rRNA gene, with a resultant sensitivity for Plasmodium falciparum of around 100 parasites per microl. Here we describe the use of mitochondrial targets for LAMP-based detection of any Plasmodium genus parasite and of P. falciparum specifically. These new targets allow routine amplification from samples containing as few as five parasites per microl of blood. Amplification is complete within 30 to 40 min and is assessed by real-time turbidimetry, thereby offering rapid diagnosis with greater sensitivity than is achieved by the most skilled microscopist or antigen detection using lateral flow immunoassays.
Polley
Spencer D
SD
Department of Clinical Parasitology, Hospital for Tropical Diseases, London Hospitals University College NHS Foundation Trust, Mortimer Market, Capper Street, London WC1E 6JB, United Kingdom. Spencer.Polley@UCLH.NHS.UK
Mori
Yasuyoshi
Y
Watson
Julie
J
Perkins
Mark D
MD
González
Iveth J
IJ
Notomi
Tsugunori
T
Chiodini
Peter L
PL
Sutherland
Colin J
CJ
D004272:DNA, Mitochondrial / Q000235:genetics*; D016054:DNA, Protozoan / Q000235:genetics*; D006801:Humans; D008288:Malaria / Q000175:diagnosis*; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D010961:Plasmodium / Q000235:genetics / Q000302:isolation & purification*; D012680:Sensitivity and Specificity; D013696:Temperature; D013997:Time Factors
D004272:DNA, Mitochondrial; D016054:DNA, Protozoan