Whole genome sequence of Seneca valley virus SVV/CH/ZZ/2016 and amplification primer thereof

A whole-genome, Seneca technology, applied in the field of molecular biology, can solve the problems of economic loss and major risks in the listing of infected pigs

Active Publication Date: 2018-06-22
JINYUBAOLING BIO PHARMA CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

From November 2014 to April 2015, there were several SVV-infected pig herds in Brazil with severe clinical morbidity and death symptoms, causing serious economic ...

Method used

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  • Whole genome sequence of Seneca valley virus SVV/CH/ZZ/2016 and amplification primer thereof
  • Whole genome sequence of Seneca valley virus SVV/CH/ZZ/2016 and amplification primer thereof
  • Whole genome sequence of Seneca valley virus SVV/CH/ZZ/2016 and amplification primer thereof

Examples

Experimental program
Comparison scheme
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Embodiment 1

[0041] Example 1. A primer combination designed for one-step RT-PCR amplification of 7 nucleotide sequence fragments of Seneca Valley virus SVV / CH / ZZ / 2016 whole genome

[0042] One of the objectives of the present invention is to obtain the whole genome sequence of Seneca Valley virus SVV / CH / ZZ / 2016, so that the molecular genetic evolution trend and prevalence of Seneca Valley virus can be more comprehensive at the level of the whole genome sequence. Understand systematically for further in-depth research.

[0043] According to the whole genome nucleotide sequence of the reference strain of Seneca Valley virus in NCBI, such as SVA / CH / 01 / 2015 (GenBank: KT321458.1), SVA / CH / 02 / 2015 (GenBank: KX173339.1), SVA / CH / DL / 01 / 2016 (GenBank: KX751944.1), SVA / CH / GXI09 / 2016 (GenBank: KY038016.1), SVA / CH / LX / 01 / 2016 (GenBank: KX751945.1), SVA / CH / ZW / 01 / 2016(GenBank:KX751946.1), SVA / HLJ / CHA / 2016(GenBank:KY419132.1), SVA / CH / FJ / 2017(GenBank:KY747510.1), SVA / CH / HN / 2017(GenBank:KY747511.1), SVA / CH / HN...

Embodiment 2

[0047] Example 2: Obtaining the whole genome sequence of Seneca Valley virus SVV / CH / ZZ / 2016

[0048] Based on the primer combination obtained in Example 1, the present invention can obtain the whole genome sequence of Seneca Valley virus SVV / CH / ZZ / 2016, and the obtaining method includes the following steps:

[0049] 1. Extract the total RNA of Seneca Valley virus SVV / CH / ZZ / 2016

[0050] Follow Axyprep TM Instructions for Body Fluid Viral DNA / RNA Miniprep Kit (purchased from AXYGEN), extract the total RNA of Seneca valley virus SVV / CH / ZZ / 2016 (preservation number: CGMCC 14886), the specific steps are as follows:

[0051] (1) Reagent preparation: Prepare isopropanol containing 1% (V / V) glacial acetic acid in advance; add a specified volume of absolute ethanol to the reagents Buffer W1A and Buffer W2 respectively. That is, add 17 mL of absolute ethanol to 24 mL of Buffer W1A; add 56 mL of absolute ethanol to 24 mL of Buffer W2.

[0052] (2) Take 200 uL of the sample to be tested (the v...

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Abstract

The invention discloses a whole genome sequence of Seneca valley virus SVV/CH/ZZ/2016 and an amplification primer thereof. The whole genome sequence of Seneca valley virus SVV/CH/ZZ/2016 is obtained by the following steps: amplifying 7 nucleotide sequence fragments (S1, S2, S3, S4, S5, S6 and S7) of the strain of SVV/CH/ZZ/2016 by a one-step RT-PCR method; performing clone sequencing on the 7 nucleotide sequence fragments; sequentially splicing, editing and correcting the DNA sequences of the 7 nucleotide sequence fragments, to finally obtain the whole genome sequence of Seneca valley virus SVV/CH/ZZ/2016. The whole genome sequence of Seneca valley virus SVV/CH/ZZ/2016 obtained in the invention is favorable for further studying the pathogenic mechanism, molecular epidemiology, reverse genetics and the like of the Seneca valley virus, and provides important data support and theoretical basis for the diagnostic reagent development, vaccine development and the like of the Seneca valley virus.

Description

Technical field [0001] The present invention belongs to the whole genome sequence of Seneca valley virus in the field of molecular biology, and particularly relates to the whole genome sequence of Seneca valley virus SVV / CH / ZZ / 2016 and its amplification primers. Background technique [0002] Seneca Valley Virus (SVV) is also called Seneca Valley Virus (SVV), Porcine Seneca Valley Virus (SVV), and Senecavirus A (SVA). SVV is a single-stranded, non-segmented, non-enveloped RNA virus, and is the only member of the genus Seneca virus, a picornavirus. SVV virus particles are typically icosahedral symmetry, with a diameter of about 27nm and a molecular weight of about 30KD. The SVV genome is about 7.2 kb in length, including two conserved non-coding regions 5'-UTR, 3'-UTR and an open reading frame (ORF), ending with a poly (A) at the 3'end. [0003] Seneca Valley virus (SVV) is a newly emerged virus that can infect piglets and sows and cause piglet death. SVV infection can cause blist...

Claims

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Application Information

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IPC IPC(8): C12N15/41C12N7/00C12N15/11C12N15/10C12R1/93
CPCC12N7/00C12N15/10C12N15/1003C07K14/005C12N2770/32021C12N2770/32022
Inventor 陈君彦王秀明魏学峰刘国英关平原陈九连范秀丽张燕红张贵刚王云凌王艳杰张宸刘建奇武瑾贤杜宇荣谢雪岑
Owner JINYUBAOLING BIO PHARMA CO LTD
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