PoSaV GIII type strain whole genome amplification primer and amplification method thereof

A technology for whole-genome amplification and amplification primers, which is applied in the field of whole-genome amplification primers and amplification of PoSaVGIII strains, and can solve problems such as the complexity of PoSaV diagnosis and control, and the difficulty of eliminating or preventing natural infection of piglets

Pending Publication Date: 2022-07-22
YUNNAN AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Nearly all PoSaV genotypes are circulating in Asia, Europe, and North America, and PoSaV diagnosis and control are becoming increasingly complex
The prevention and control of the disease mainly relies on comprehensive measures, and it is difficult to eliminate or prevent natural infection of piglets in pig herds

Method used

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  • PoSaV GIII type strain whole genome amplification primer and amplification method thereof
  • PoSaV GIII type strain whole genome amplification primer and amplification method thereof
  • PoSaV GIII type strain whole genome amplification primer and amplification method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment

[0059] 1 Materials and methods

[0060] 1.1 Materials

[0061] 1.1.1 Sample source

[0062] Pig Sapporo virus-positive disease materials were isolated from fecal samples of pigs with diarrhea, provided by the Animal Infectious Disease Laboratory, School of Veterinary Medicine, Yunnan Agricultural University. The positive disease materials were numbered YNJD158 and YNAN326.

[0063] 1.1.2 Main reagents

[0064] TRipure Reagent total RNA extraction reagent was purchased from Beijing Biotech Co., Ltd.; columnar plasmid DNA mini-extraction kit and gel recovery kit were purchased from Sangon Bioengineering (Shanghai) Co., Ltd.; Easyscript RT / RI Enzyme MIX, 2× Es Reaction MIX, 2×TranTaq HiFi PCR Super Mix II were purchased from Beijing Quanshijin Biotechnology Co., Ltd.; pMD18-T Vector, E.coli DH5α Competent Cells, restriction enzymes, etc. were purchased from Bao Bio (Dalian) Co., Ltd.; other Reagents were provided by the Animal Infectious Diseases Laboratory, School of Veterina...

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Abstract

The invention discloses a PoSaV GIII type strain whole genome amplification primer and an amplification method thereof. The amplification method comprises the following steps: 1) RNA extraction, 2) whole genome segmentation amplification, 3) disease sample sequencing, and 4) whole genome sequence analysis to analyze genetic variation characteristics of a disease sample; the invention provides a research basis for the development of the PoSaV vaccine, and provides a scientific basis for effectively preventing and controlling the prevalence of the disease in regions.

Description

technical field [0001] The present invention relates to the field of biotechnology, in particular to a PoSaV GIII type strain whole genome amplification primer and an amplification method thereof. Background technique [0002] Sapporo virus (Sapovirus, SaV), also known as Zaru virus, belongs to Calicivirus family, Sapporovirus genus, and is considered to be one of the viral pathogens causing acute gastroenteritis worldwide. found in the baby room. In 1980, American researchers used an electron microscope to observe the diarrhea intestinal contents of a 27-day-old suckling pig and identified the first porcine Sapporo virus (Porcine Sapovirus, PoSaV), which was named Cowden strain, Zhang Wen in 2008. In 2009, Huang Zebin and other investigations showed that the virus existed in pig herds in Hunan and Guangdong provinces, but the prevalence of the disease in Yunnan pig herds has not been reported. [0003] SaV divides Sapporo virus into five genotypes based on the gene sequen...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/70C12Q1/686C12N15/11C12R1/93
CPCC12Q1/701C12Q1/686C12Q2521/107C12Q2565/125
Inventor 李小兵尹革芬赵谦毕峻宇刘应华刘霄
Owner YUNNAN AGRICULTURAL UNIVERSITY
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