Toxoplasma gondii: sensitive and rapid detection of infection by loop-mediated isothermal amplification (LAMP) method.
Loop-mediated isothermal amplification (LAMP) method amplifies DNA with high specificity, sensitivity and rapidity. In this study, we used a conserved sequence in the 200- to 300-fold repetitive 529 bp gene of Toxoplasma gondii to design primers for LAMP test. Detection limit of T. gondii LAMP assay with the primers is 1 pg/microL of T. gondii DNA, which was evaluated using 10-fold serially diluted DNA of cultured parasites. Furthermore, LAMP and conventional PCR methods were applied for amplification of the T. gondii DNA extracted from the lymph nodes taken from pigs which were suspected to be Toxoplasma infection. As a result, 76.9% (70/91) and 85.7% (78/91) of the samples were positive on PCR and LAMP analyzes, respectively. Therefore, the LAMP has a potential to be applied as an alternative molecular diagnostic tool for detection of T. gondii infection from veterinary samples. This is the first study, which applies the LAMP method to diagnose Toxoplasma from veterinary samples.
Zhang
Houshuang
H
National Research Center for Protozoan Diseases, Obihiro University of Agriculture and Veterinary Medicine, Inada-cho, Obihiro, Hokkaido 080-8555, Japan.
Thekisoe
Oriel M M
OM
Aboge
Gabriel O
GO
Kyan
Hisako
H
Yamagishi
Junya
J
Inoue
Noboru
N
Nishikawa
Yoshifumi
Y
Zakimi
Satoshi
S
Xuan
Xuenan
X
D000818:Animals; D002522:Chlorocebus aethiops; D016054:DNA, Protozoan / Q000032:analysis*; D021141:Nucleic Acid Amplification Techniques / Q000379:methods / Q000592:standards / Q000662:veterinary*; D016133:Polymerase Chain Reaction / Q000662:veterinary; D012680:Sensitivity and Specificity; D013047:Specific Pathogen-Free Organisms; D013552:Swine; D013553:Swine Diseases / Q000175:diagnosis / Q000469:parasitology; D014122:Toxoplasma / Q000235:genetics / Q000302:isolation & purification*; D014124:Toxoplasmosis, Animal / Q000175:diagnosis*; D014709:Vero Cells
D016054:DNA, Protozoan