BACKGROUND Parasites of the family Spirorchiidae cause disease and mortality in marine and freshwater turtles; two species, Hapalotrema mistroides and Neospirorchis sp., are reported in the resident population of loggerhead turtles of the Mediterranean Sea, with the first being the most widespread. In vivo diagnosis of spirorchidiasis can represent a challenge in guaranteeing prompt control and treatment of the disease and is currently limited to copromicroscopy. The aim of this study was the development of a real time PCR assay with TaqMan probe for the detection of H. mistroides infection in the blood of live loggerhead turtles, Caretta caretta, hospitalized in rehabilitation centres. Its potential use for in vivo diagnosis is explored. RESULTS The developed real time PCR successfully detected H. mistroides DNA from both positive controls and experimental blood samples of live loggerhead sea turtles, showing good specificity, sensitivity and good reaction efficiency. Two out of three turtles which had demonstrated positivity at copromicroscopy also tested positive to this blood assay; DNA of H. mistroides was detected within the blood of one sea turtle, which tested negative for copromicroscopy. CONCLUSIONS This study describes a specific and rapid molecular assay to detect H. mistroides infection from live sea turtles and highlights for the first time the presence of DNA of this species in turtle blood samples. Since this assay is able to detect low amounts of the parasitic free DNA in blood samples, its application could be helpful for in vivo diagnosis of H. mistroides infection as well as for epidemiological purposes.

中文翻译：

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活海龟乌龟Caretta caretta血液样本中的Hapalotrema雾状病毒（Digenea：Spirorchiidae）感染的分子诊断的初步研究。

背景技术螺旋藻科的寄生虫在海洋和淡水龟中引起疾病​​和死亡。据报道，在地中海的龟常居种群中，有两个物种，Hapalotrema mistroides和Neospirorchis sp。，其中第一个是分布最广的。螺旋藻的体内诊断在保证疾病的快速控制和治疗方面可能是一个挑战，目前仅限于共显微技术。这项研究的目的是开发一种用TaqMan探针进行实时PCR检测的方法，用于检测在康复中心住院的活（Caretta caretta）血液中的类螺旋体感染。探索了其在体内诊断中的潜在用途。结果开发的实时荧光定量PCR成功检测到H。活的logger海龟的阳性对照和实验血样中的槲寄生DNA均显示出良好的特异性，敏感性和良好的反应效率。在辅助显微镜下显示阳性的三分之二的乌龟也对该血液检测呈阳性。在一只海龟的血液中检测到了雾状疱疹的DNA，该样品经共显微镜检查呈阴性。结论这项研究描述了一种特异性和快速的分子检测方法，可检测活海龟感染的类螺旋体，并首次突出显示了龟血样本中该物种的DNA的存在。由于该检测方法能够检测到血液样本中的少量游离寄生DNA，因此其应用可能有助于体内诊断类螺旋体感染和流行病学研究。