PCR method for quickly amplifying whole genome sequence of porcine circovirus type 2 and application thereof

A porcine circovirus and whole genome technology, applied in the field of molecular biology, can solve problems such as rapid amplification and acquisition of unfavorable whole genome sequences, homologous recombination, etc.

Inactive Publication Date: 2015-12-02
INST OF ANIMAL HEALTH GUANGDONG ACADEMY OF AGRI SCI
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  • Abstract
  • Description
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AI Technical Summary

Problems solved by technology

However, when amplifying the whole genome of porcine circovirus type 2 in these studies, the amplification primers used are often multiple primers, which often leads to artificial homologous recombination. Rapid Amplification and Acquisition of the Whole Genome Sequence of Cyclovirus Type 2

Method used

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  • PCR method for quickly amplifying whole genome sequence of porcine circovirus type 2 and application thereof
  • PCR method for quickly amplifying whole genome sequence of porcine circovirus type 2 and application thereof
  • PCR method for quickly amplifying whole genome sequence of porcine circovirus type 2 and application thereof

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Experimental program
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Embodiment Construction

[0018] The present invention will be further described below in conjunction with specific embodiments, and its operation steps will be further clarified.

[0019] Optimized embodiment:

[0020] (1) Viral DNA extraction

[0021] Take buffalo and Rattus norvegicus samples that were identified as positive for porcine circovirus type 2 by PCR, grind, homogenate, dilute at 1:2, freeze and thaw repeatedly 3 times, and carry out according to the virus DNA extraction kit (Axygen Company) DNA extraction, finally, use 50-100 μL RNase-free double-distilled water for DNA elution, and store in a -20°C refrigerator;

[0022] (2) Primer design and synthesis

[0023] Through the alignment of the whole genome sequences of multiple genotypes of porcine circovirus type 2, a pair of primers (chemically synthesized by Suzhou Jinweizhi Biotechnology Co., Ltd.) were designed using Primer Premier 5.0 software. The specific primer sequences are shown in Table 1. "F" stands for forward primer and "R...

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Abstract

The invention discloses a PCR method for quickly amplifying a whole genome sequence of porcine circovirus type 2 and application thereof. Primers used for amplifying the whole genome sequence the porcine circovirus type 2 include primers shown as SEQ ID No.1 and SEQ ID No.2. The PCR method is high in specificity and sensitivity, capable of quickly and effectively amplifying target segments and conducive to studying genetic evolution, molecular epidemiology and infectious clone of the porcine circovirus type 2 with different animal sources.

Description

technical field [0001] The patent of the present invention belongs to the field of molecular biology, and specifically relates to a rapid amplification method of porcine circovirus type 2 whole genome sequence and its application. Background technique [0002] Porcine circovirus type 2 is the main pathogen of porcine circovirus disease. Clinically, it often leads to piglet multisystem failure syndrome, porcine dermatitis and nephrotic syndrome, reproductive disorders in sows, interstitial pneumonia, etc., which seriously endangers pigs. Healthy development of the industry (Zhai et al., 2014, Virology Journal, 11:88). Porcine circovirus has two types: porcine circovirus type 1 and porcine circovirus type 2, and porcine circovirus type 2 has multiple genotypes, such as PCV2a, PCV2b, PCV2c, PCV2d and PCV2e. Porcine circovirus type 1 is not pathogenic to pigs, but porcine circovirus type 2 is pathogenic to pigs and is the focus of scientific research. Porcine circovirus is ubi...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/70C12Q1/68C12R1/93
CPCC12Q1/6869C12Q1/701C12Q2531/113
Inventor 翟少伦魏文康温肖会吕殿红
Owner INST OF ANIMAL HEALTH GUANGDONG ACADEMY OF AGRI SCI
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