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A detection method for grass carp hemorrhagic virus (GCHV) based on a reverse transcription polymerase chain reaction
Li, J; Wang, TH; Yi, YL; Liu, HQ; Lu, RH; Chen, HX
1997
Source PublicationDISEASES OF AQUATIC ORGANISMS
Volume29Issue:1Pages:7-12
AbstractA rapid, sensitive and highly specific detection method for grass carp hemorrhagic virus (GCHV) based on a reverse transcription-polymerase chain reaction (RT-PCR) has been developed. Two pairs of PCR primers were synthesized according to the cloned cDNA sequences of the GCHV-861 strain. For each primer combination, only one specific major product was obtained when amplification was performed by using the genomic dsRNA of GCHV-861 strain. The lengths of their expected products were 320 and 223 bp, respectively. No products were obtained when nucleic acids other than GCHV-861 genomic RNA were used as RT-PCR templates. To assess the sensitivity of the method, dilutions of purified GCHV-861 dsRNA total genome (0.01 pg up to 1000 pg) were amplified and quantities of as little as 0.1 pg of purified dsRNA were detectable when the amplification product was analyzed by 1.5% agarose gel electrophoresis. This technique could detect GCHV-861 not only in infected cell culture fluids, but also in infected grass carp Ctenopharyngodon idellus and rare minnow Gobiocypris rarus with or without hemorrhagic symptoms. The results show that the RT-PCR amplification method is useful for the direct detection of GCHV.
KeywordHemorrhagic Virus Of Grass Carp (Gchv) Virus Detection Reverse Transcription Polymerase Chain Reaction
DepartmentCHINESE ACAD SCI,INST HYDROBIOL,WUHAN 430072,PEOPLES R CHINA
Subject AreaFisheries ; Veterinary Sciences
Indexed BySCI
WOS IDWOS:A1997XE32700002
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Document Type期刊论文
Identifierhttp://ir.ihb.ac.cn/handle/152342/10266
Collection期刊论文
Recommended Citation
GB/T 7714
Li, J,Wang, TH,Yi, YL,et al. A detection method for grass carp hemorrhagic virus (GCHV) based on a reverse transcription polymerase chain reaction[J]. DISEASES OF AQUATIC ORGANISMS,1997,29(1):7-12.
APA Li, J,Wang, TH,Yi, YL,Liu, HQ,Lu, RH,&Chen, HX.(1997).A detection method for grass carp hemorrhagic virus (GCHV) based on a reverse transcription polymerase chain reaction.DISEASES OF AQUATIC ORGANISMS,29(1),7-12.
MLA Li, J,et al."A detection method for grass carp hemorrhagic virus (GCHV) based on a reverse transcription polymerase chain reaction".DISEASES OF AQUATIC ORGANISMS 29.1(1997):7-12.
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