挪威龙虾（Nephrops norvegicus）及其他甲壳类动物中血红哈卡藻（Hematodinium spp.）的分子检测

Molecular detection of Hematodinium spp. in Norway lobster Nephrops norvegicus and other crustaceans.

作者信息

Small H J, Neil D M, Taylor A C, Atkinson R J A, Coombs G H

机构信息

Division of Environmental and Evolutionary Biology, Institute of Biomedical and Life Sciences, University of Glasgow, UK.

出版信息

Dis Aquat Organ. 2006 Apr 6;69(2-3):185-95. doi: 10.3354/dao069185.

DOI:10.3354/dao069185

PMID:16724562

Abstract

The Norway lobster Nephrops norvegicus (L.) from the coastal waters of Scotland is seasonally infected by a parasitic dinoflagellate of the genus Hematodinium. Methods used to detect infection include a morphological index (pleopod diagnosis) and several immunoassays. The present study describes the development and application of a set of Hematodinium-specific polymerase chain reaction (PCR) primers and DNA probes based on Hematodinium ribosomal DNA (rDNA). In the PCR assay, a diagnostic band of 380 bp was consistently amplified from total genomic DNA isolated from Hematodinium-infected N. norvegicus. The sensitivity of the assay was 1 ng DNA, which is equivalent to 0.6 parasites. The primer pair also detected Hematodinium DNA in preparations of the amphipod Orchomene nanus, indicating that the amphipod may be infected with the same Hematodinium sp. infecting N. norvegicus. DNA probes detected Hematodinium parasites in heart, hepatopancreas and gill tissues from N. norvegicus, and hepatopancreas and gill tissues from Carcinus maenas, confirming Hematodinium infection in the latter.

摘要

来自苏格兰沿海水域的挪威龙虾（Nephrops norvegicus (L.)）会季节性地感染一种名为血卵涡鞭虫属（Hematodinium）的寄生性双鞭毛虫。用于检测感染的方法包括形态学指标（腹足诊断）和几种免疫测定法。本研究描述了一组基于血卵涡鞭虫核糖体DNA（rDNA）的血卵涡鞭虫特异性聚合酶链反应（PCR）引物和DNA探针的开发与应用。在PCR测定中，从感染血卵涡鞭虫的挪威龙虾分离的总基因组DNA中始终能扩增出一条380 bp的诊断条带。该测定的灵敏度为1 ng DNA，相当于0.6个寄生虫。该引物对还在双足虾Orchomene nanus的制剂中检测到了血卵涡鞭虫DNA，这表明双足虾可能感染了与感染挪威龙虾相同的血卵涡鞭虫物种。DNA探针在挪威龙虾的心脏、肝胰腺和鳃组织以及平背蜞的肝胰腺和鳃组织中检测到了血卵涡鞭虫寄生虫，证实了后者存在血卵涡鞭虫感染。