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Detection of Herpesvirus anguillae (Ang HV ‐1) in European eel Anguilla anguilla (L.) originating from northern Poland—assessment of suitability of selected diagnostic methods
Author(s) -
Nguyen Tuan Thuc,
Jin Yeonhwa,
Kiełpińska Jolanta,
Bergmann Sven M.,
Lenk Matthias,
Panicz Remigiusz
Publication year - 2017
Publication title -
journal of fish diseases
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.819
H-Index - 85
eISSN - 1365-2761
pISSN - 0140-7775
DOI - 10.1111/jfd.12689
Subject(s) - biology , nested polymerase chain reaction , polymerase chain reaction , virology , virus , real time polymerase chain reaction , microbiology and biotechnology , gene , genetics
The Community Action Plan requests EU member states to implement measures that ensure the recovery of the severely depleted European eel stocks. One of the main threats is posed by Anguillid herpesvirus 1 (Ang HV ‐1) leading to increased mortality in both wild and farmed eels. Following recommendations of the OIE to minimize the risk of obtaining false‐negative results, the main aim of the study was to optimize diagnostic methods for Ang HV ‐1 detection using conventional PCR , nested PCR and in situ hybridization assay. While 53.3% of the individual organ samples were tested positive for Ang HV ‐1 by PCR , the additional virus analysis via nested PCR revealed that the actual prevalence was 93.3%. In the cell cultivation passages, a cytopathic effect was hardly found in the first two rounds. In the third passage onto cell cultures, a lytic CPE was detected. The identification and confirmation of the viruses obtained from cell cultures as well as directly from the organ tissues were proceeded by PCR , nested PCR and sequencing of the PCR products. While no positive signal was detectable in the first round by PCR using samples from the third cell culture passages, the nested PCR provided weak but visible positive signals.