College of Veterinary Medicine, South China Agricultural University, Guangzhou, Guangdong Province 510642, People's Republic of China.
Parasitol Res. 2010 May;106(6):1379-83. doi: 10.1007/s00436-010-1812-3. Epub 2010 Mar 16.
The fish-borne clonorchiasis caused by the oriental liver fluke Clonorchis sinensis is endemic in a number of countries with over 35 million people being infected globally. Rapid and accurate detection of C. sinensis in its intermediate host fish is important for the control and prevention of clonorchiasis in areas where the disease is endemic. In the present study, we established a loop-mediated isothermal amplification (LAMP) approach for the sensitive and rapid detection of C. sinensis metacercariae in fish. The specificity and sensitivity of primers designed from the C. sinensis cathepsins B3 gene were evaluated, and specific amplification products were obtained with C. sinensis, while no amplification products were detected with DNA of related trematodes, demonstrating the specificity of the assay. The LAMP assay was proved to be 100 times more sensitive than a conventional polymerase chain reaction for detection of C. sinensis. The established LAMP assay provides a useful tool for the rapid and sensitive detection of C. sinensis in fish, which has important implications for the effective control of human clonorchiasis.
由东方肝吸虫 Clonorchis sinensis 引起的鱼类肝吸虫病在多个国家流行,全球有超过 3500 万人感染。快速准确地检测中间宿主鱼类中的 C. sinensis 对于控制和预防该疾病流行地区的肝吸虫病非常重要。在本研究中,我们建立了一种环介导等温扩增 (LAMP) 方法,用于敏感和快速检测鱼类中的 C. sinensis 囊蚴。从 C. sinensis 组织蛋白酶 B3 基因设计的引物的特异性和灵敏度进行了评估,并且仅用 C. sinensis 的 DNA 获得了特异性的扩增产物,而用相关吸虫的 DNA 则未检测到扩增产物,证明了该测定方法的特异性。LAMP 检测比常规聚合酶链反应检测 C. sinensis 的灵敏度高 100 倍。所建立的 LAMP 检测方法为快速灵敏地检测鱼类中的 C. sinensis 提供了一种有用的工具,这对于有效控制人类肝吸虫病具有重要意义。