Discrimination of Opisthorchis viverrini from Haplorchis taichui using COI sequence marker
Discrimination of Opisthorchis viverrini from Haplorchis taichui using COI sequence marker
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DOI：
10.1016/j.actatropica.2007.05.006
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发表时间：
2007-07-01
期刊：
影响因子：
2.7
通讯作者：
Waikagul, Jitra
中科院分区：
文献类型：
--
作者：
Thaenkham, Urusa;Visetsuk, Kasidis;Waikagul, Jitra
本研究旨在基于线粒体细胞色素c氧化酶亚基I （COI）基因对泰国常见的两种鱼源性吸虫（viverrini Opisthorchis和Haplorchis taichui）的感染进行区分。设计的引物（COI- ov - hap - F&R引物）在不同发育阶段（成虫、囊蚴和卵）扩增出了O. viverrini和H. taichui的部分COI片段，灵敏度较高。采用低基因组DNA浓度（约10(-4)ng），在50℃和56℃的退火温度下，分别获得了O. viverrini和H. taichi的PCR扩增子。在50℃条件下，也获得了华支睾吸虫和太水睾吸虫的COI片段，但其敏感性低于O. viverrini。在56℃条件下，只扩增出太水嗜血杆菌，并与pumilio、H. yogokawai、O. viverrini和C. sinensis区分。在50 ~ 56℃范围内，pumilio和H. yogokawai的PCR扩增产物特异性较低，敏感性较低。采用聚合酶链反应- rflp方法对产弧菌、中华弧菌和太水弧菌的遗传性状进行了比较。PCR-RFLP图谱可用于诊断猪瘟弧菌和太水弧菌混合感染，以及在鱼类宿主感染流行病学调查中检测猪瘟弧菌、中华弧菌和太水弧菌的囊蚴。(c) 2007 Elsevier B.V.版权所有
This study aimed to discriminate infections of two common fish-borne trematodes in Thailand, Opisthorchis viverrini from Haplorchis taichui, based on mitochondrial cytochrome c oxidase subunit I (COI) gene. Designed primers (COI-OV-Hap F&R primers) amplified partial COI fragments of O. viverrini and H. taichui with high sensitivity in different developmental stages (adult, metacercaria, and egg). Polymerase chain reaction (PCR) amplicons were generated with low genomic DNA concentration (approximate to 10(-4) ng)of O. viverrini and H. taichiii at 50 and 56 degrees C annealing temperatures, respectively. At 50 degrees C, COI fragments of Clonorchis sinensis and H. taichui were also obtained, but this was less sensitive than O. viverrini. At 56 degrees C, only H. taichui could be amplified and discriminated from H. pumilio, H. yogokawai, O. viverrini, and C. sinensis. Between 50 and 56 degrees C, the PCR amplicons of H. pumilio and H. yogokawai were amplified with low specificity and low sensitivity. The genetic characters among O. viverrini, C. sinensis, and H. taichui were distinguished by PCR-RFLP method. The PCR-RFLP profiles might be useful for diagnosing mixed O. viverrini and H. taichui infections in endemic areas, and for detecting metacercariae of O. viverrini, C. sinensis and H. taichui in epidemiological surveys of infections in fish hosts. (c) 2007 Elsevier B.V. All rights reserved.