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Development and Evaluation of a Loop-mediated Isothermal Amplification(LAMP) Assay for Rapid Detection of Chinese Giant Salamander ranavirus

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Asian Herpetological Research 2015年第1期6卷 59-65页

作者： Yi GENG Xingxing LIU Yan ZHOU Kaiyu WANG Xi PENG Zhijun ZHONG Xiaoli HUANG Defang CHEN College of Veterinary Medicine Sichuan Agricultural University Department of Aquaculture Sichuan Agricultural University

Loop-mediated isothermal ampliifcation （LAMP） is a novel nucleic acid diagnostic method that can amplify rapidly a target template under isothermal conditions. In this study, a LAMP assay for rapid detection of Chinese giant salamander ranavirus（CGSRV） was developed from culture isolates and clinical samples. The LAMP assay was developed by designing one set of four speciifc primers, targeting the major capsid protein （MCP） gene of CGSRV. Reaction time and temperature were optimal for 40 min at 62°C. The developed LAMP assay is speciifc and highly sensitive for CGSRV detection, the detection limit could reach about 5 copies of cloned viral genomic fragments. The sensitivity of the LAMP assay was about 1000 and 10-fold higher than that of both conventional and nested PCR, respectively. The LAMP ampliifcation produces a typical ladder-like pattern of products on an agarose gel that can be visually inspected after addition of ethidium bromide. The LAMP assay was evaluated further with clinical samples, and the results indicated the suitability and simplicity of the test as a rapid diagnostic tool for the detection of CGSRV.

关键词： CGSRV Loop-mediated isothermal ampliifcation （LAMP） ranavirus Chinese giant salamander

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Pathological Changes in Andrias davidianus Infected with Chinese Giant Salamander ranavirus

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Asian Herpetological Research 2016年第4期7卷 258-264页

作者： Yi GENG Matthew James GRAY Kaiyu WANG Defang CHEN Ping OUYANG Xiaoli HUANG Changliang HE Zhijun ZHONG Zexiao YANG College of Veterinary Medicine Sichuan Agricultural University Center for Wildlife Health University of Tennessee Department of Aquaculture Sichuan Agricultural University

Chinese giant salamander ranavirus （CGSRV） is an emerging pathogen in captive populations of the Chinese giant salamander （Andrias davidianus）. We processed 140 morbid Chinese giant salamanders from seven captive breeding populations over five years, and describe the disease associated with CGSRV infection. The most common gross signs were significant swelling of the legs and coelomic cavity, erythema of the legs and ventrum in juveniles; cutaneous erosions and ulcerations in adults, particularly the limbs and the head; and marked petechial or ecchymotic hemorrhages of the internal organs, particularly the liver, spleen and kidney. Histological examination showed degeneration, necrosis, and inflammation in many organs, particularly in the organs where hemorrhage was observed. There was evidence of eosinophilic inclusion bodies in degenerated and necrotic cells. We identified virus particles and empty capsids without viral nucleoid in the inclusion bodies using electron microscopy. Virus particles were hexagonal or round shape, and appeared in paracrystalline arrays, aggregates, or singly. All enveloped viral particles were 140-160 nm. Polymerase chain reaction followed by sequencing verified that the virus particles were CGSRV. These results collectively support that CGSRV was the etiologic agent responsible for these mass die-offs of the Chinese giant salamander. The pathology described herein will be useful in diagnosing cases of ranaviral disease caused by CGSRV, and provide evidence that this pathogen is a significant threat to the Chinese giant salamander.

关键词： pathology CGSRV Andrias davidianus ranavirus China

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Susceptibility of farmed juvenile giant grouper Epinephelus lanceolatus to a newly isolated grouper iridovirus（genus ranavirus）

Susceptibility of farmed juvenile giant grouper Epinephelus ...

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2014年中国水产学会学术年会

作者： Chao Peng Hongling Ma Youlu Su Weigeng Wen Juan Feng Zhixun Guo Lihua Qiu Key Laboratory of South China Sea Fishery Resources Exploitation & Utilization Ministry of Agriculture The South China Sea Fisheries Research InstituteChinese Academy of Fishery Sciences College of Fisheries and Life Science Shanghai Ocean University

A ranavirus was isolated from the diseased farmed groupers, Epinephelus hybrids（blotchy rock cod, Epinephelus fuscoguttatus ♀× giant grouper, E. lanceolatus ♂）, in Sanya, Hainan, in July 2013. In this study, susceptibility of E. lanceolatus to GIV-R was determined by intraperitoneally injection. The cumulative mortality reached to 81% at 5 day post infection. Histologically, severe degeneration with massive pycnotic nuclei in spleen and kidney tissues was observed,and some small-size inclusion body-bearing cells（IBCs） existed in spleen. Hemorrhage and infiltration of inflammatory cells were presented in gill, liver and heart along with tissue degeneration and necrosis of varying severity. The results of immunohistochemistry analysis showed that the strongest immunolabellings were obtained from the kidney and spleen tissues, while intermediate intensity signals were observed in the heart, stomach, gill and liver tissues, and the weakest signals were obtained from the intestine and brain, but no signal was obtained in eyes. In surviving fish, abnormal hypertrophic cells were observed in both splenic corpuscle and renal corpuscle, while no hypertrophic cell was observed in the other parts of spleen and kidney tissues. Moreover, immunolabellings only stained the hypertrophic cells in splenic corpuscle and renal corpuscle.

关键词： Epinephelus lanceolatus Histopathology Immunohistochemistry ranavirus

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Molecular basis of pathogenesis of emerging viruses infecting aquatic animals

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Aquaculture and Fisheries 2018年第1期3卷 1-5页

作者： Lang Gui V.Gregory Chinchar Qiya Zhang College of Fisheries and Ljfe Science Shanghai Ocean University.Shanghai 201306China Department of Microbiolbgy.University of Miss tsippi Medical Center 2500 North State StreetJacksonMS 39216USA State Key Laboratory of Freshwater Ecology and Botechnology Institute of Hydrobiology.Chinese Acodemy of SciencesWuhan 430072China

Aquatic vertebrates are very abundant in the world,and they are of tremendous importance in providing global food security and nutrition.However,emergent and resurgent viruses,such as ranavirus(e.g.,Rana grylio virus,RGV and Andriasd avidianus ranavirus,ADRV),herpesvirus(e.g.,Carassius carassius herpesvirus,CaHV),reovirus(e.g.,grass carp reovirus 109,GCRV-109,Scophthal musmaximus reovirus,SMReV and Micropterus salmoides reovirus,MsReV),and rhabdovirus(e.g.,Siniper cachuatsi rhabdovirus,SCRV and Scophthal musmaximus rhabdovirus,SMRV)can cause severe diseases in aquaculture animals and wild lower vertebrates,such as frogs,giant salamanders,fish,and so on.Here,we will briefly describe the symptoms produced by the aforementioned viruses and the molecular basis of the virus ehost interactions.This manuscript aims to provide an overview of viral diseases in lower vertebrates with an emphasis on visible symptomatic manifestations and pathogenesis.

关键词： ranavirus Herpesvirus Reovirus Rhabdovirus Aquatic animal

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