Rognlie M C, Dimke K L, Knapp S E
Montana State University, Bozeman 59717-0360.
J Parasitol. 1994 Oct;80(5):748-55.
Fasciola hepatica, the common bile duct fluke, is an economically important parasite of domestic livestock. Current research interest is directed toward an understanding of the parasite's biology at the intermediate host level. To permit study of seasonal transmission patterns and parasite/intermediate host interactions, a fasciolid-specific assay has been developed to detect infected snail vectors. This assay uses the reverse transcriptase-polymerase chain reaction (RT-PCR) to amplify specifically a region of F. hepatica small-subunit rRNA, followed by hybridization to an F. hepatica-specific probe. The assay does not cross-react with 2 trematodes outside of the Fasciolidae but does detect Fascioloides magna rRNA. Sequence alignment with additional small-subunit rRNAs shows Fasciolopsis buski would also cross-react with the assay. The detection limit of the assay is 10 fg of fluke total RNA with 5 micrograms of snail RNA added as background. Additionally, the assay detects individual infected snails immediately after miracidial exposure and throughout the parasite's development period.
肝片吸虫，即常见的胆管吸虫，是家畜中一种具有重要经济意义的寄生虫。当前的研究兴趣集中在了解该寄生虫在中间宿主层面的生物学特性。为了便于研究季节性传播模式以及寄生虫与中间宿主的相互作用，已开发出一种针对片形吸虫的检测方法来检测受感染的蜗牛媒介。该检测方法利用逆转录聚合酶链反应（RT-PCR）特异性扩增肝片吸虫小亚基rRNA的一个区域，随后与肝片吸虫特异性探针杂交。该检测方法不会与片形科以外的2种吸虫发生交叉反应，但能检测到巨片形吸虫的rRNA。与其他小亚基rRNA的序列比对显示，布氏姜片吸虫也会与该检测方法发生交叉反应。该检测方法的检测限为10 fg的吸虫总RNA，加入5微克蜗牛RNA作为背景。此外，该检测方法在毛蚴暴露后立即以及在寄生虫的整个发育期间都能检测到单个受感染的蜗牛。
