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Loop-mediated isothermal amplification reaction primer for detecting porcine parvovirus II

A loop-mediated isothermal, parvovirus technology, applied in microorganisms, microorganism-based methods, biochemical equipment and methods, etc., can solve problems such as inability to be widely used in grass-roots laboratories, and improve the field usability and applicability. , the effect of high sensitivity

Inactive Publication Date: 2013-05-08
FUJIAN AGRI & FORESTRY UNIV
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Problems solved by technology

[0003] For the detection of porcine parvovirus type 2 based on the molecular level, specific primers are usually designed according to the characteristics of the virus gene for PCR amplification, which can timely and accurately carry out the differential diagnosis of pathogens. However, ordinary PCR and fluorescent quantitative PCR with higher sensitivity need to use precision Instruments have high technical requirements for experimenters, etc., and cannot be widely used in grassroots laboratories

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  • Loop-mediated isothermal amplification reaction primer for detecting porcine parvovirus II
  • Loop-mediated isothermal amplification reaction primer for detecting porcine parvovirus II

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Embodiment 1

[0018] 1. Design of primers for LAMP experiments

[0019] Download all porcine parvovirus type 2 gene sequences registered in GenBank and the porcine parvovirus type 2 gene sequences detected by our laboratory from the field pig farms in our province for analysis and comparison, and select the conserved region fragment of the front part of the VP1 gene (GenBankJX101461) , use the online website (http: / / primerexplorer.jp / e / ) designed by LAMP primers to design primers, including 1 pair of outer primers and 1 pair of inner primers. For specific sequences, see SEQ ID NO.1, SEQ ID NO.2, SEQ ID NO. ID NO.3 and SEQ ID NO.4.

[0020] 2. Virus strain

[0021] Porcine parvovirus, porcine circovirus, porcine pseudorabies virus, and swine fever virus were all purchased from China Veterinary Microbiological Culture Collection and Management Center. The positive DNA of porcine parvovirus type 2 was detected and preserved on-site by the pig farm in our province (Fujian Province).

[0022]...

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Abstract

The invention belongs to the filed of animal molecular virology. The invention discloses a loop-mediated isothermal amplification reaction primer for detecting porcine parvovirus II (PPV II). According to a method, two pairs of primers consisting of outer primers F3 and B3 and inner primers FIP and BIP are designed according to a forepart conserved sequence of a porcine parvovirus II VP1 gene. The specific sequences of the two pairs of primers are shown as SEQ ID NO. 1, SEQ ID NO. 2, SEQ ID NO. 3 and SEQ ID NO. 4. A loop-mediated isothermal amplification method for detecting porcine parvovirus II (PPV II) is established according to the designed primer. No cross reaction occurs in porcine ordinary infectious disease viruses according to the detection method. The detection method has strong on-site practicability without requiring expensive instrument and equipment, high accuracy, strong specificity and high sensitivity.

Description

technical field [0001] The invention relates to a loop-mediated isothermal amplification reaction primer for detecting porcine parvovirus type 2 (PPV2), belonging to the field of animal molecular virology. Background technique [0002] Porcine parvovirus 2 (PPV2) was first detected in a pig with hepatitis E in Myanmar in 2001. Since then, it has been detected in pigs suffering from "high fever" in southern my country (China) in 2006-2007. Recent studies of the disease in Hungary and the US military have reported. At present, studies on the pathogenicity of PPV2 are relatively small, and the pathogenic mechanism has not yet been clarified. Therefore, it is of great significance to establish a simple and practical detection method for the prevention and control of porcine parvovirus type 2. [0003] For the detection of porcine parvovirus type 2 based on the molecular level, specific primers are usually designed according to the characteristics of the virus gene for PCR amp...

Claims

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Application Information

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IPC IPC(8): C12Q1/70C12Q1/68C12N15/11C12R1/93
Inventor 李家玉张奇林志华
Owner FUJIAN AGRI & FORESTRY UNIV
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