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Primer combination for detecting porcine circovirus 3 by aid of loop-mediated isothermal amplification method

A ring-mediated isothermal, porcine circovirus technology, which is applied in the field of animal infectious disease detection to achieve the effect of improving field usability, reducing the possibility of pollution and good applicability

Active Publication Date: 2017-11-03
INST OF ANIMAL HUSBANDRY & VETERINARY FUJIAN ACADEMY OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there is no method for loop-mediated isothermal amplification (LAMP) reaction, but there are corresponding LAMP detection methods for other types of viruses in this genus, such as PCV1 and PCV2

Method used

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  • Primer combination for detecting porcine circovirus 3 by aid of loop-mediated isothermal amplification method
  • Primer combination for detecting porcine circovirus 3 by aid of loop-mediated isothermal amplification method

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Experimental program
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Embodiment 1

[0018] 1. Experimental methods and steps

[0019] 1.1. Design of primers for LAMP experiments

[0020] According to all the gene sequences of porcine circovirus type 3 registered in GenBank and the genome sequences of porcine circovirus type 3 detected by our laboratory from the field pig farms in our province (Fujian Province), we used molecular biology software to analyze and compare , select the conserved region of the genome of porcine circovirus type 3 (GenBank accession number KT869077), and use the online website for LAMP primer design (https: / / primerexplorer.jp / lampv5 / index.html) to design primers, including a pair of outer primers and 1 pair of internal primers, see SEQ ID NO.1, SEQ ID NO.2, SEQ ID NO.3 and SEQ ID NO.4 for specific sequences.

[0021] 1.2. Virus strains

[0022] Porcine parvovirus (PPV), porcine circovirus type 1 (PCV1), porcine circovirus type 2 (PCV2), porcine pseudorabies virus (PRV), porcine reproductive and respiratory syndrome virus (PRRSV) an...

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Abstract

The invention belongs to the field of animal infectious disease detection, and discloses a primer combination for detecting porcine circovirus 3 (PCV3) by the aid of a loop-mediated isothermal amplification (LAMP) method. The loop-mediated isothermal amplification method includes designing two pairs of primers including outer primers F3 and B3 and inner primers FIP and BIP according to genome conserved sequences of the porcine circovirus 3. The particular sequences are shown as SEQ ID NO.1, SEQ ID NO.2, SEQ ID NO.3 and SEQ ID NO.4. The loop-mediated isothermal amplification method for detecting the porcine circovirus 3 is established according to the designed primers. The primer combination has the advantages that cross reaction with pathogens of common porcine infectious diseases can be prevented; expensive instruments and equipment can be omitted, the loop-mediated isothermal amplification method is convenient to use in situ and high in practicality; the primer combination is high in accuracy, specificity and sensitivity.

Description

technical field [0001] The invention relates to a primer set used for detecting porcine circovirus type 3 loop-mediated isothermal amplification reaction, and belongs to the field of animal infectious disease detection. Background technique [0002] Porcine circoviruses (Porcine circoviruses, PCV) are single-stranded circular DNA viruses with a genome length of about 1.7 kb, and are one of the smallest animal DNA viruses. Two types of PCV have been identified, circovirus type 1 (PCV1) and circovirus type 2 (PCV2). PCV1 was first identified as a contaminant in PK cell cultures in 1974 and is not pathogenic in pigs. PCV2 was first reported in 1998. It can cause porcine circovirus associated diseases (PCVAD) in pigs under clinical conditions, mainly causing piglet multisystem failure syndrome, pneumonia, porcine dermatitis, nephrotic syndrome and Reproductive disorders, mainly manifested as respiratory, urinary, intestinal, lymphatic, cardiovascular, nervous, reproductive sys...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/70C12Q1/68C12N15/11
CPCC12Q1/6844C12Q1/701C12Q2600/16C12Q2531/119C12Q2537/143
Inventor 陈如敬陈秋勇吴学敏车勇良王晨燕周伦江严山王隆柏魏宏
Owner INST OF ANIMAL HUSBANDRY & VETERINARY FUJIAN ACADEMY OF AGRI SCI