We isolated a strain of lymphocystis disease virus （LCDV） from Japanese flounder （Paralichthys olivaceus） cultured in northern China. Based on published sequences of major capsid protein （MCP） gene of LCDV-cn （...We isolated a strain of lymphocystis disease virus （LCDV） from Japanese flounder （Paralichthys olivaceus） cultured in northern China. Based on published sequences of major capsid protein （MCP） gene of LCDV-cn （GenBank： AF126405）, we designed two primer sets P1/P2 and P3/P4. We then used one-step or nested PCR and in-situ hybridization （ISFI） to detect LCDV and identify the target tissues or cells in infected Japanese flounder. The PCR products were positive in purified viral supematant, skin nodules, gut, gill, kidney, spleen, stomach, heart, and liver of Japanese flounder. We compared the DNA sequence with 14 MCP nucleotide sequences from GenBank, including Megalocytivirus （OFIV and RSIV）, lridovirus （CzlV and W/V）, Ranavirus （TFV and FV3）, and Lymphocystivirus （8 LCDV）. Based on the alignment, we confirmed the PCR product was from Lymphocystivirus （GenBank accession number DQ279090 （LCDV-HD））. Using ISH, we noted the presence of LCDV in the skin nodules, gut, gill, spleen, stomach, and heart of spontaneously infected Japanese flounders. We successfully amplified LCDV fragments from Schlegel＇s black rockfish （Sebastes schlegeli Higendorf）, redwing sea robin （Lepidotrigla microptera Gtinther） and turbot （Scophthalmus maximus） using the one-step and nested PCR, suggesting the target genes can be widely detected in fish using this method.展开更多
Iridovirus infection often causes death and considerable economic losses in the aquaculture industry. This research applies the co-agglutination method that is fast, cheap and accurate in confirming the diagnosis of t...Iridovirus infection often causes death and considerable economic losses in the aquaculture industry. This research applies the co-agglutination method that is fast, cheap and accurate in confirming the diagnosis of the cause of an outbreak of illness caused by iridovirus in the field, so that remedial action can be taken quickly and appropriately to minimize the impact of wider losses. Samples were taken from the grouper and pomfret star farms that are experiencing outbreaks of infectious diseases in the months from May to August 2015, Tanjungpinang, Indonesia. The sick and allegedly attacked by iridovirus samples showed abnormal swimming clinical symptoms, weakness and the swollen spleen. The swollen spleen of sick fish created suspension in phosphate buffered saline （PBS） with pH 7.2, and then centrifuged at 8,000 rpm for I0 rain. The supernatant after centrifuge was used as the test sample. On a clean object glass, 50 μL of the supematant was treated with 50 μL kit co-agglutination pre-prepared. The positive results were shown by the agglutination reaction after 10-15 rain, while as a negative control, PBS was reacted with co-agglutination kit that looked homogeneous （no agglutination）. It was showed that the grouper （Epinepkelus sp.） on four farms and pomfret star （Thracinotus blochii） on one farm that experienced an outbreak of infectious disease in Tanjungpinang showed positively infected iridovirus. The same positive iridovirus result was also demonstrated by examination using polymerase chain reaction （PCR） at 570 bp. So, the causative agent of plague on grouper and pomfret star was iridovirus. In addition, the co-agglutination test based on serology is more quick, cheap and accurate.展开更多
基金Supported by the National Natural Science Foundation of China (No 30771648)the National High Technology Research and Development Program of China (863 Program) (No 2006AA100306)
文摘We isolated a strain of lymphocystis disease virus （LCDV） from Japanese flounder （Paralichthys olivaceus） cultured in northern China. Based on published sequences of major capsid protein （MCP） gene of LCDV-cn （GenBank： AF126405）, we designed two primer sets P1/P2 and P3/P4. We then used one-step or nested PCR and in-situ hybridization （ISFI） to detect LCDV and identify the target tissues or cells in infected Japanese flounder. The PCR products were positive in purified viral supematant, skin nodules, gut, gill, kidney, spleen, stomach, heart, and liver of Japanese flounder. We compared the DNA sequence with 14 MCP nucleotide sequences from GenBank, including Megalocytivirus （OFIV and RSIV）, lridovirus （CzlV and W/V）, Ranavirus （TFV and FV3）, and Lymphocystivirus （8 LCDV）. Based on the alignment, we confirmed the PCR product was from Lymphocystivirus （GenBank accession number DQ279090 （LCDV-HD））. Using ISH, we noted the presence of LCDV in the skin nodules, gut, gill, spleen, stomach, and heart of spontaneously infected Japanese flounders. We successfully amplified LCDV fragments from Schlegel＇s black rockfish （Sebastes schlegeli Higendorf）, redwing sea robin （Lepidotrigla microptera Gtinther） and turbot （Scophthalmus maximus） using the one-step and nested PCR, suggesting the target genes can be widely detected in fish using this method.
文摘Iridovirus infection often causes death and considerable economic losses in the aquaculture industry. This research applies the co-agglutination method that is fast, cheap and accurate in confirming the diagnosis of the cause of an outbreak of illness caused by iridovirus in the field, so that remedial action can be taken quickly and appropriately to minimize the impact of wider losses. Samples were taken from the grouper and pomfret star farms that are experiencing outbreaks of infectious diseases in the months from May to August 2015, Tanjungpinang, Indonesia. The sick and allegedly attacked by iridovirus samples showed abnormal swimming clinical symptoms, weakness and the swollen spleen. The swollen spleen of sick fish created suspension in phosphate buffered saline （PBS） with pH 7.2, and then centrifuged at 8,000 rpm for I0 rain. The supernatant after centrifuge was used as the test sample. On a clean object glass, 50 μL of the supematant was treated with 50 μL kit co-agglutination pre-prepared. The positive results were shown by the agglutination reaction after 10-15 rain, while as a negative control, PBS was reacted with co-agglutination kit that looked homogeneous （no agglutination）. It was showed that the grouper （Epinepkelus sp.） on four farms and pomfret star （Thracinotus blochii） on one farm that experienced an outbreak of infectious disease in Tanjungpinang showed positively infected iridovirus. The same positive iridovirus result was also demonstrated by examination using polymerase chain reaction （PCR） at 570 bp. So, the causative agent of plague on grouper and pomfret star was iridovirus. In addition, the co-agglutination test based on serology is more quick, cheap and accurate.
