Young N D, Dyková I, Nowak B F, Morrison R N
School of Aquaculture, Tasmanian Aquaculture and Fisheries Institute, University of Tasmania and Aquafin CRC, Launceston, Tasmania, Australia.
J Fish Dis. 2008 Apr;31(4):285-95. doi: 10.1111/j.1365-2761.2008.00903.x.
The recent description of Neoparamoeba perurans as an aetiological agent of amoebic gill disease (AGD) advanced our understanding of the condition and has forced a re-evaluation of methods used for the diagnosis of AGD. Currently, there are no tools available that are both specific for N. perurans and suitable for a routine diagnostic procedure. Therefore, in this study we describe an assay to detect N. perurans. The assay, which utilizes PCR to amplify the N. perurans 18S rRNA gene, was shown to be specific and highly sensitive. Neoparamoeba perurans was detected in both gill samples and primary isolates of non-cultured gill-derived amoebae obtained during necropsy or biopsy from AGD-affected Atlantic salmon, Salmo salar L. The PCR-based assay provides a simple, flexible tool that will be a useful addition to the diagnostic repertoire for AGD. It may also be used for the genotypic screening of trophozoites during culture and could facilitate further epidemiological and ecological studies of AGD.
最近，秘鲁新帕拉变形虫被描述为阿米巴鳃病（AGD）的病原体，这加深了我们对该病的理解，并促使人们重新评估用于AGD诊断的方法。目前，尚无既对秘鲁新帕拉变形虫具有特异性又适用于常规诊断程序的工具。因此，在本研究中，我们描述了一种检测秘鲁新帕拉变形虫的检测方法。该检测方法利用聚合酶链反应（PCR）扩增秘鲁新帕拉变形虫的18S核糖体RNA（rRNA）基因，结果显示具有特异性和高度敏感性。在对受AGD影响的大西洋鲑（Salmo salar L.）进行尸检或活检时获得的鳃样本以及未培养的鳃源性变形虫的原始分离物中均检测到了秘鲁新帕拉变形虫。基于PCR的检测方法提供了一种简单、灵活的工具，将成为AGD诊断方法中的一项有用补充。它还可用于培养过程中滋养体的基因型筛选，并有助于进一步开展AGD的流行病学和生态学研究。
