Triple polymerase chain reaction (PCR) detecting primer group, reagent kit and method for diagnosing grass carp reovirus
A technology of reovirus and detection kit, which is applied in the detection primer set, triple PCR detection of different subtypes of grass carp reovirus, and the field of kits, which can solve the complicated operation, the inability to quickly diagnose the virus, and the unfavorable grass carp hemorrhagic disease Prevention and other problems, to achieve the effect of cost savings
Inactive Publication Date: 2012-07-04
PEARL RIVER FISHERY RES INST CHINESE ACAD OF FISHERY SCI
View PDF1 Cites 12 Cited by
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
[0003] Virus isolation, electron microscope observation, and nucleic acid band type analysis are still commonly used methods to detect grass carp reovirus, but these methods are relatively complicated to operate and cannot quickly diagnose the virus, which is not conducive to timely and effective prevention of grass carp haemorrhagic disease
Method used
the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View moreImage
Smart Image Click on the blue labels to locate them in the text.
Smart ImageViewing Examples
Examples
Experimental program
Comparison scheme
Effect test
Embodiment 1
[0035] 1. Collection of test strains and disease materials
[0036] Koi herpesvirus type 3 (KHV), infectious hematopoietic necrosis virus (IHNV), largemouth bass iridescent virus (LMBV), infectious spleen and kidney necrosis virus (ISKNV), GCRV HZ08 strain, GCRV JX09-01, GCRV JX10 -01 strain, GCRV JX09-02, and GCRV 104 strain were isolated and preserved in this experiment, and GSRV was purchased from ATCC. The disease material used to isolate the pathogen is collected from the visceral tissue of the fish suffering from the corresponding fish disease. The disease material tissue is collected aseptically and stored in the phosphate buffer solution containing double antibody. Each tube contains 25 ~ 30ml buffer solution, -80℃ save.
[0037] Primer design and synthesis
[0038] All the gene sequences of GCRV existing in GENBANK were analyzed, and a 532bp conserved sequence of the S6 gene of JX09-01 and 873 strains, a 196bp conserved sequence of the S6 gene of HZ08 and GD10 str...
the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More PUM
Login to View More Abstract
The invention discloses a triple polymerase chain reaction (PCR) detecting primer group, a reagent kit and a method for diagnosing grass carp reovirus. The method disclosed by the invention is a triple RR-PCR detecting method capable of simultaneously detecting three kinds of subtypes of the grass carp reovirus: HZ08 strain types, 104 strain types and JX09-01 strain types, the subtypes of the grass carp reovirus can be identified when the grass carp reovirus is detected, the valuable time is obtained for the providing of prevention and control measures in a targeted way and more precise treatment according to diseases, and the important significance is realized on the effective control of grass carp bleeding diseases. When being applied to the detection of the grass carp reovirus, the primer group and the reagent kit disclosed by the invention have the advantages of accuracy, flexibility, high speed, cost saving and the like.
Description
technical field [0001] The invention belongs to the field of virus molecular biology detection, in particular to triple PCR detection of different subtypes of grass carp reovirus, including a detection primer set, a kit and a method. Background technique [0002] Grass carp reovirus (GCRV) belongs to the genus Aquatic Reovirus, a new member of Reoviridae, and the first fish virus isolated from mainland China. The virus mainly causes haemorrhagic disease in freshwater cultured grass carp species in China, Vietnam, Myanmar and other Asian countries, and the mortality rate can be as high as 60%. The virus is widespread, harmful, high in mortality, and has a long onset season, which seriously threatens fishery production. Grass carp reovirus has a double-layer capsid, the average diameter of the virion is 60 nm-70 nm, icosahedral symmetry, no envelope, and the genome consists of 11 segmented double-stranded RNAs. At present, nearly 20 isolates have been reported, including GCR...
Claims
the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More Application Information
Patent Timeline
Login to View More IPC IPC(8): C12Q1/70C12Q1/68C12N15/11
Inventor 曾伟伟王庆吴淑勤王英英张乐生石存斌
Owner PEARL RIVER FISHERY RES INST CHINESE ACAD OF FISHERY SCI