Species-specific loop-mediated isothermal amplification (LAMP) for diagnosis of trypanosomosis.
In this study, we developed loop-mediated isothermal amplification (LAMP) for the specific detection of both animal and human trypanosomosis using primer sets that are designed from 5.8S rRNA-internal transcribed spacer 2 (ITS2) gene for Trypanosoma brucei gambiense, 18S rRNA for both T. congolense and T. cruzi, and VSG RoTat 1.2 for T. evansi. These LAMP primer sets are highly sensitive and are capable of detecting down to 1 fg trypanosomal DNA, which is equivalent to approximately 0.01 trypanosomes. LAMP is a rapid and simple technique since it can be carried out in 1 h and requires only a simple heating device for incubation. Therefore, LAMP has great potential of being used for diagnosis of trypanosomosis in the laboratory and the field, especially in countries that lack sufficient resources needed for application of molecular diagnostic techniques.
Thekisoe
Oriel M M
OM
National Research Center for Protozoan Diseases, Obihiro University of Agriculture and Veterinary Medicine, Obihiro, Hokkaido 080 8555, Japan.
Kuboki
Noritaka
N
Nambota
Andrew
A
Fujisaki
Kozo
K
Sugimoto
Chihiro
C
Igarashi
Ikuo
I
Yasuda
Jun
J
Inoue
Noboru
N
D000818:Animals; D001483:Base Sequence; D017931:DNA Primers; D016054:DNA, Protozoan / Q000235:genetics; D021903:DNA, Ribosomal Spacer / Q000235:genetics; D008969:Molecular Sequence Data; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D012680:Sensitivity and Specificity; D013045:Species Specificity; D014345:Trypanosoma / Q000235:genetics* / Q000302:isolation & purification*; D014352:Trypanosomiasis / Q000175:diagnosis* / Q000469:parasitology*
D017931:DNA Primers; D016054:DNA, Protozoan; D021903:DNA, Ribosomal Spacer