State Key Laboratory of Agricultural Microbiology, College of Veterinary Medicine, Huazhong Agricultural University, Hubei, Wuhan, 430070, PR China.
Vet Parasitol. 2010 Jul 15;171(1-2):159-62. doi: 10.1016/j.vetpar.2010.02.046. Epub 2010 Mar 3.
Theileria is a widespread, intraerythrocytic tick-borne protozoan parasite. Loop-mediated isothermal amplification (LAMP) is a novel nucleic acid amplification method offering rapid, accurate and cost-effective diagnosis of infectious diseases. In this study, the LAMP method was developed for detecting Theileria sergenti. Four primers were designed from six distinct regions of the target gene, a 33-kDa major piroplasm surface protein (p33) gene in T. sergenti. The specificity assay showed it was specific for T. sergenti whilst the LAMP was able to detect a parasitemia level of 0.000002% which was more sensitive than conventional PCR. 154 field samples from water buffalo and 159 field samples from cattle were analyzed using the LAMP method. About 60.4% (96/159) of cattle samples were positive by LAMP, compared to 30.0% (46/154) of water buffalo samples that were positive. Compared with conventional PCR, the LAMP method exhibited higher detection abilities than conventional PCR. All the results indicated that the LAMP assay is a simple and convenient diagnostic tool for theileriosis.
泰勒虫是一种广泛存在的、红细胞内寄生的蜱传原生动物寄生虫。环介导等温扩增（LAMP）是一种新型的核酸扩增方法，为传染病的快速、准确和经济有效的诊断提供了可能。本研究中，建立了一种检测瑟氏泰勒虫的 LAMP 方法。根据瑟氏泰勒虫 33kDa 主要血孢子表面蛋白（p33）基因的 6 个不同区域设计了 4 条引物。特异性试验显示，该方法具有特异性，可检测到 0.000002%的疟原虫血症水平，比常规 PCR 更敏感。使用 LAMP 方法分析了来自水牛的 154 个现场样本和来自牛的 159 个现场样本。结果显示，LAMP 检测牛样本的阳性率为 60.4%（96/159），而水牛样本的阳性率为 30.0%（46/154）。与常规 PCR 相比，LAMP 方法具有更高的检测能力。所有结果表明，LAMP 检测方法是一种简单方便的泰勒虫病诊断工具。
