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Primer set for detecting infectious spleen and kidney necrosis virus and siniperca chuatsi rhabdovirus and kit

A technology of spleen and kidney necrosis virus and rhabdovirus, which is applied in the direction of recombinant DNA technology, microbial measurement/testing, biochemical equipment and methods, etc., can solve the problems of cost increase, detection time extension, delay of mandarin fish, etc., and achieve saving Cost, the effect of simplifying the operation procedure

Pending Publication Date: 2019-05-17
PEARL RIVER FISHERY RES INST CHINESE ACAD OF FISHERY SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The PCR detection methods of ISKNV and SCRV have been widely established, but the general PCR reaction can only detect one corresponding pathogenic virus at a time, and the phenomenon of mixed infection of two viruses usually occurs in fish, which requires multiple detection and analysis. Sequencing can determine the strain type, which prolongs the detection time, increases the cost, and delays the control of ISKNV and SCRV in mandarin fish

Method used

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  • Primer set for detecting infectious spleen and kidney necrosis virus and siniperca chuatsi rhabdovirus and kit
  • Primer set for detecting infectious spleen and kidney necrosis virus and siniperca chuatsi rhabdovirus and kit
  • Primer set for detecting infectious spleen and kidney necrosis virus and siniperca chuatsi rhabdovirus and kit

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Experimental program
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Effect test

Embodiment 1

[0029] 1. Design of primers

[0030] Analyze the sequences of the MCP gene of ISKNV and the N gene of SCRV in GENBANK, design the first pair of primers ISKNV-1300MCP-F / R for the MCP gene of ISKNV; design the second pair of primers ISKNV-550- for the conserved region of the MCP gene of ISKNV F / R, the sequence length is 550bp; the third pair of primers SCRV-QY-N-F / R is designed for the N gene of SCRV, and the sequence length is 1290bp; the fourth pair of primers SCRV-280-F / R is designed for the conserved region of the N gene of SCRV , the sequence length is 280bp. The primers were synthesized by Guangzhou Qingke Biotechnology Co., Ltd., and the sequences are as follows:

[0031]

[0032]

[0033] After the primers were synthesized, the specificity of the primers was verified by single PCR and double PCR amplification by mixing two plasmid templates and two pairs of primers designed according to the conserved regions. PCR reaction system 25 μL: take P-N 0.5 μL, P-MCP 1 μL...

Embodiment 2

[0049] Embodiment 2 double PCR identification method application

[0050] Using the 20 suspected ISKNV and SCRV samples collected and preserved in major mandarin fish farms such as Foshan and Dongguan in 2018 by our laboratory, we used the previously established double PCR method for detection.

[0051] 20 suspected ISKNV and SCRV samples kept in the laboratory were detected by ISKNV single PCR, SCRV single PCR, ISKNV and SCRV double PCR detection at the same time, and the effects of the double PCR and single PCR detection methods established by the present invention were compared.

[0052] The result is as Figure 8-10 as shown, Figure 8 Among them, 3 SCRVs were detected positive, and the positive rate was 15%; Figure 9 Among them, 11 cases were positive for ISKNV, and the positive rate was 55%; Figure 10 Among them, 2 samples were positive for mixed infection of SCRV and ISKNV, and the positive rate was 10%.

[0053] Visible single PCR detection method can only detect...

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Abstract

The invention provides a primer set for detecting infectious spleen and kidney necrosis virus and siniperca chuatsi rhabdovirus. The primer set includes two pairs of primers, and the sequences of thefirst pair of primers are shown as SEQ ID NO: 3 and SEQ ID NO: 4, the sequences of the second pair of primers are shown as SEQ ID NO: 7, SEQ ID NO: 8. According to the primer set, the infectious spleen and kidney necrosis virus and siniperca chuatsi rhabdovirus can be quickly detected simultaneously, operation procedures can be simplified, and the cost is reduced.

Description

technical field [0001] The invention belongs to the technical field of detection of aquatic disease microorganisms, and in particular relates to a primer set and a kit for detecting infectious spleen and kidney necrosis virus and mandarin fish rhabdovirus. Background technique [0002] Infectious spleen and kidney necrosis virus (ISKNV) and mandarin fish rhabdovirus (Siniperca chuatsi rhabdovirus, SCRV) are the main viral pathogens affecting mandarin fish aquaculture in recent years. Fish diseases have the consequences of high morbidity, high infectivity and high lethality, which bring huge economic losses to the aquaculture industry. [0003] Infectious spleen-kidney necrosis virus (ISKNV) is a double-stranded DNA virus belonging to the Iridoviridae genus Swollen cell. ISKNV is mainly prevalent in sea and freshwater cultured fish such as grouper and mandarin fish, and the lethality rate to mandarin fish can be as high as 100%. Siniperca mandarin rhizome virus (SCRV) was f...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/70C12Q1/686C12N15/11
Inventor 梁红茹范芷仪蔡秀珠李宁求林强付小哲刘礼辉黄志斌牛银杰
Owner PEARL RIVER FISHERY RES INST CHINESE ACAD OF FISHERY SCI
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