一种用于诊断人类、猫和鱼类中华支睾吸虫感染的特异性聚合酶链反应检测方法。
A specific PCR assay for the diagnosis of Clonorchis sinensis infection in humans, cats and fishes.
作者信息
Huang Si-Yang, Tang Jian-Dong, Song Hui-Qun, Fu Bao-Quan, Xu Min-Jun, Hu Xu-Chu, Zhang Han, Weng Ya-Biao, Lin Rui-Qing, Zhu Xing-Quan
机构信息
State Key Laboratory of Veterinary Etiological Biology, Key Laboratory of Veterinary Parasitology of Gansu Province, Lanzhou Veterinary Research Institute, CAAS, Lanzhou, Gansu Province 730046, PR China.
出版信息
Parasitol Int. 2012 Mar;61(1):187-90. doi: 10.1016/j.parint.2011.07.010. Epub 2011 Jul 14.
Clonorchiasis caused by Clonorchis sinensis is a fish-borne parasitic disease which is endemic in a number of countries. Using the sequences of the internal transcribed spacers (ITS-1 and ITS-2) of nuclear ribosomal DNA (rDNA) of C. sinensis as genetic markers, a pair of C. sinensis-specific primers was designed and used to establish a specific PCR assay for the diagnosis of C. sinensis infection in humans, cats and fish. This approach allowed the specific identification of C. sinensis after optimizing amplification conditions, with no amplicons being amplified from related heterogeneous DNA samples, and sequencing of amplicons confirmed the identity of the sequences amplified. The detection limit of this assay was 1.03 pg of adult C. sinensis, 1.1 metacercariae per gram of fish filet, and a single egg in human and cat feces. The PCR assay should provide a useful tool for the diagnosis and molecular epidemiological investigation of clonorchiasis in humans and animals.
华支睾吸虫引起的华支睾吸虫病是一种在许多国家流行的食源性寄生虫病。以华支睾吸虫核糖体DNA（rDNA）的内转录间隔区（ITS-1和ITS-2）序列作为遗传标记，设计了一对华支睾吸虫特异性引物，并用于建立一种特异性PCR检测方法，用于诊断人、猫和鱼体内的华支睾吸虫感染。在优化扩增条件后，该方法能够特异性鉴定华支睾吸虫，未从相关异源DNA样本中扩增出扩增子，扩增子测序证实了扩增序列的同一性。该检测方法的检测限为成年华支睾吸虫1.03 pg、每克鱼片1.1个囊蚴以及人和猫粪便中的单个虫卵。该PCR检测方法应为人类和动物华支睾吸虫病的诊断和分子流行病学调查提供一种有用的工具。
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