一种用于检测蚁属和毛蚁属蚂蚁体内枝双腔吸虫的DNA探针

A DNA probe for the detection of Dicrocoelium dendriticum in ants of Formica spp. and Lasius spp.

作者信息

Heussler V, Kaufmann H, Glaser I, Ducommun D, Müller C, Dobbelaere D

机构信息

University of Bern, Institute of Animal Pathology, Switzerland.

出版信息

Parasitol Res. 1998 Jun;84(6):505-8. doi: 10.1007/s004360050437.

DOI:10.1007/s004360050437

PMID:9660142

Abstract

Repetitive DNA sequences present in the genome of Dicrocoelium dendriticum were identified by hybridization of genomic DNA that had been digested with different restriction enzymes with 32P-labeled genomic D. dendriticum DNA. DNA fragments containing repetitive sequences were isolated from PstI-digested D. dendriticum DNA and were subcloned into a plasmid vector. Plasmids containing repetitive sequences were identified by colony hybridization. One of these plasmids, designated Ddr-IV, was isolated and used as a probe in further studies. Ddr-IV is specific for D. dendriticum since it does not hybridize to DNA isolated from other trematodes. In addition, Ddr-IV was capable of detecting D. dendriticum metacercariae in ants (Formica cunicularia, F. rufibarbis, and Lasius sp.), which act as second intermediate hosts in the parasite's life cycle. Since metacercariae constitute the infectious stage of the parasite for grazing animals, Ddr-IV will provide a useful tool for epidemiology studies of dicrocoeliosis.

摘要

通过用不同限制性内切酶消化的基因组 DNA 与经 32P 标记的枝双腔吸虫基因组 DNA 杂交，鉴定了枝双腔吸虫基因组中存在的重复 DNA 序列。从经 PstI 消化的枝双腔吸虫 DNA 中分离出含有重复序列的 DNA 片段，并将其亚克隆到质粒载体中。通过菌落杂交鉴定含有重复序列的质粒。其中一个质粒，命名为 Ddr-IV，被分离出来并用作进一步研究的探针。Ddr-IV 对枝双腔吸虫具有特异性，因为它不与从其他吸虫分离的 DNA 杂交。此外，Ddr-IV 能够检测蚂蚁（穴居蚁、红褐林蚁和毛蚁属）体内的枝双腔吸虫囊蚴，这些蚂蚁在寄生虫的生命周期中充当第二中间宿主。由于囊蚴是寄生虫对放牧动物的感染阶段，Ddr-IV 将为双腔吸虫病的流行病学研究提供一个有用的工具。