title: Comparison of loop-mediated isothermal amplification (LAMP) and real-time PCR method targeting a 529-bp repeat element for diagnosis of toxoplasmosis.
abstract: Loop-mediated isothermal amplification (LAMP) is a simple method that can amplify DNA with high specificity, sensitivity, and rapidity. In this study, we compared the performance of LAMP and real-time PCR assays for diagnosis of toxoplasmosis. We designed a real-time PCR assay targeting a 529 bp element repeated 200-300 times in the Toxoplasma gondii genome. The detection limits of the LAMP and real-time PCR assays were 10 fg/μL and 1 fg/μL of T. gondii DNA, respectively. Conventional PCR, LAMP, and real-time PCR methods were applied to detect T. gondii DNA in blood samples from 284 pigs and 292 sheep. Positive results were obtained with 0.4%, 3.2%, and 4.2% of the pig samples and 3.8%, 17.1%, and 17.8% of the sheep samples with conventional PCR, LAMP, and real-time PCR analyses, respectively. The real-time PCR assay provided the most sensitive diagnosis of toxoplasmosis, but the LAMP assay has potential as an alternative tool for detection of T. gondii in the field.
authors: Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences, No. 518 Ziyue Road, Minhang District, Shanghai 200241, China.
authors: Zhibing
authors: Z
authors: Lin
authors: Yanlei
authors: Y
authors: Zhang
authors: Houshuang
authors: H
authors: Zhang
authors: Yongzhi
authors: Y
authors: Zhou
authors: Jie
authors: J
authors: Cao
authors: Jinlin
authors: J
authors: Zhou
mesh_terms: D000818:Animals; D021141:Nucleic Acid Amplification Techniques / Q000662:veterinary*; D060888:Real-Time Polymerase Chain Reaction / Q000662:veterinary*; D012680:Sensitivity and Specificity; D012756:Sheep; D012757:Sheep Diseases / Q000097:blood / Q000175:diagnosis / Q000469:parasitology; D013552:Swine; D013553:Swine Diseases / Q000097:blood / Q000175:diagnosis / Q000469:parasitology; D014122:Toxoplasma / Q000235:genetics* / Q000302:isolation & purification*; D014124:Toxoplasmosis, Animal / Q000175:diagnosis*