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Preparation method for standard sample of infectious hematopoietic organ necrosis virus molecule

A technology for hematopoietic organ necrosis and molecular standard samples, which is applied in the field of preparation of virus molecular standard samples, can solve problems such as difficulty in ensuring laboratory quality control, difficulty in maintaining stability and uniformity, time-consuming and labor-intensive, etc., and saves time in the preparation process Labor saving, good uniformity, guaranteed quality control effect

Inactive Publication Date: 2012-08-15
DALIAN NATIONALITIES UNIVERSITY
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  • Abstract
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  • Claims
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Problems solved by technology

However, since the virus is an RNA virus, its RNA is extremely easy to be degraded. It is not only time-consuming and labor-intensive to extract the RNA virus in each laboratory, but it is also difficult to maintain its stability and uniformity, and it is difficult to ensure the quality control of the laboratory.

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  • Preparation method for standard sample of infectious hematopoietic organ necrosis virus molecule
  • Preparation method for standard sample of infectious hematopoietic organ necrosis virus molecule
  • Preparation method for standard sample of infectious hematopoietic organ necrosis virus molecule

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Embodiment Construction

[0040] Target fragments were selected according to the fragments in OIE and "GB / T 15805.2-2008 Fish Quarantine Methods Part 2: Infectious Hematopoietic Necrosis Virus (IHNV)" for laboratory quality control.

[0041] a. Extraction of infectious hematopoietic necrosis virus RNA

[0042] Add 600 μl of Trizol lysate and 200 μl of infectious hematopoietic necrosis virus cell culture to the centrifuge tube. The culture was purchased from the China Center for Type Culture Collection (GDV012), and a positive control and a negative control were set at the same time; add 200 μl of chloroform and shake to mix. , centrifuge at 10 000 g for 15 min; absorb 500 μl of the supernatant and add it to 500 μl of isopropanol, invert and mix repeatedly, and centrifuge at 10 000 g for 15 min; gently discard the supernatant, place it upside down on absorbent paper, dry the liquid, add Wash with 600 μl of 75% ethanol upside down, centrifuge at 10 000 g for 10 min; discard the supernatant, place it upsi...

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Abstract

The invention discloses a preparation method for a standard sample of an infectious hematopoietic organ necrosis virus molecule. The preparation method is characterized by including following steps: extracting an infectious hematopoietic organ necrosis virus RNA (ribonucleic acid); performing RT-PCR (reverse transcription-polymerase chain reaction); purifying target genes; performing connection transformation of target segments; recycling plasmid; performing in vitro transcription; performing RNA purification after transcription; and performing phenol chloroform extraction. The preparation method is time-saving and labor-saving in a preparation process, the standard sample of prepared infectious hematopoietic organ necrosis virus ribonucleic acid is high in stability and fine in uniformity, standard samples can be provided for infectious hematopoietic organ necrosis virus detection researches, medicine researches, application researches and the like, so that comparison of different laboratory results is realized, and quality control of laboratories is guaranteed.

Description

technical field [0001] The invention relates to a preparation method of a virus molecular standard sample, in particular to a molecular standard sample of infectious hematopoietic organ necrosis virus which saves time and effort, has high stability and good uniformity. Background technique [0002] Infectious haematopoietic necrosis virus belongs to the genus Novirhabdovirus of the Infectious haematopoietic necrosis virus (IHNV) family, and aquatic animal viruses of the same genus include Hirame rhabdovirus (HRV), snakehead bomb Snakehead rhabdovirUS (SRV) and Viral haemorrhagic septicaemia (VHSV). IHNV is the representative species of this genus. The representative strain of IHNV is WRAC strain (ATCC code is VR-1392). The floating density in cesium sulfate is 1.59 g / ml, unstable to heat, acid and ether. [0003] Infectious haematopoietic necrosis (IHN) is a serious infectious disease in salmon, trout and other fish. Due to the great harm caused by the virus, OIE listed it...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/63C12N15/47
Inventor 杜雄伟李叶
Owner DALIAN NATIONALITIES UNIVERSITY
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