Loop-mediated isothermal amplification (LAMP) assays for detection of Theileria parva infections targeting the PIM and p150 genes.
We have developed two loop-mediated isothermal amplification (LAMP) assays for the detection of Theileria parva, the causative agent of East Coast fever (ECF), an economically important cattle disease in eastern, central and southern Africa. These assays target the polymorphic immunodominant molecule (PIM) and p150 LAMP genes. The primer set for each gene target consists of six primers, and each set recognises eight distinct regions on the target gene to give highly specific detection of T. parva. The detection limit of each primer set is 1fg, which is equivalent to one copy of the PIM and p150 T. parva genes. These PIM and p150 LAMP primer sets amplify DNA of T. parva isolates from cattle and buffalo from different countries including Kenya, South Africa, Tanzania, Rwanda, Uganda and Burundi, indicating their ability to detect T. parva from different countries. With the advantages of simplicity, rapidity and cost effectiveness, these LAMP assays are good candidates for molecular epidemiology studies and for monitoring control programs in ECF-endemic, resource poor countries.
Thekisoe
Oriel M M
OM
National Research Center for Protozoan Diseases, Obihiro University of Agriculture and Veterinary Medicine, Obihiro, Hokkaido 080-8555, Japan.
Rambritch
Natasha E
NE
Nakao
Ryo
R
Bazie
Raoul S
RS
Mbati
Peter
P
Namangala
Boniface
B
Malele
Imna
I
Skilton
Robert A
RA
Jongejan
Frans
F
Sugimoto
Chihiro
C
Kawazu
Shin-Ichiro
S
Inoue
Noboru
N
D000818:Animals; D000953:Antigens, Protozoan / Q000235:genetics*; D002020:Buffaloes; D002063:Burundi; D002417:Cattle; D002418:Cattle Diseases / Q000175:diagnosis* / Q000469:parasitology; D017931:DNA Primers; D016054:DNA, Protozoan / Q000032:analysis / Q000235:genetics / Q000302:isolation & purification; D007630:Kenya; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D015800:Protozoan Proteins / Q000235:genetics*; D012432:Rwanda; D012680:Sensitivity and Specificity; D013019:South Africa; D013636:Tanzania; D016797:Theileria parva / Q000235:genetics / Q000302:isolation & purification*; D013801:Theileriasis / Q000175:diagnosis* / Q000469:parasitology; D013997:Time Factors; D014454:Uganda
D000953:Antigens, Protozoan; D017931:DNA Primers; D016054:DNA, Protozoan; D015800:Protozoan Proteins; C095023:antigen PIM, Theileria parva