Loop-mediated isothermal amplification primer and method for detecting type 2 porcine circovirus

A porcine circovirus, ring-mediated isothermal technology, applied in biochemical equipment and methods, microbial measurement/testing, DNA/RNA fragments, etc., can solve the problem of unsatisfactory specificity and sensitivity, prone to false positives, To detect problems such as low sensitivity, achieve the effect of shortening the reaction time, good specificity, and speeding up the reaction process

Inactive Publication Date: 2017-02-01
XUCHANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] The second technical problem to be solved by the present invention is that the method for detecting type II porcine circovirus in the prior art has low detection sensitivity, unsatisfactory specificity and sensitivity, cumbersome operation process, prone to false positive phenomenon, detection Therefore, it provides a method for detecting type Ⅱ porcine circovirus with high detection sensitivity, good specificity and sensitivity, simple operation and high detection rate.

Method used

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  • Loop-mediated isothermal amplification primer and method for detecting type 2 porcine circovirus
  • Loop-mediated isothermal amplification primer and method for detecting type 2 porcine circovirus
  • Loop-mediated isothermal amplification primer and method for detecting type 2 porcine circovirus

Examples

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Embodiment 1

[0028] Example 1 Design of loop-mediated isothermal amplification primers for detecting type Ⅱ porcine circovirus

[0029] In this embodiment, the loop-mediated isothermal amplification primers for detecting type II porcine circovirus include a first outer primer, a second outer primer, a first inner primer, and a second inner primer; wherein, the first outer primer ( hereinafter referred to as F3), having the sequence structure shown in SEQ ID No.1; the second outer primer (hereinafter referred to as B3), having the sequence structure shown in SEQ ID No.2; the first inner primer ( hereinafter referred to as FIP) has a sequence structure as shown in SEQ ID No.3; the second internal primer (hereinafter referred to as BIP) has a sequence structure as shown in SEQ ID No.4. Its specific sequence structure and its position on the ORF1 gene for detecting type II porcine circovirus are shown in the table below:

[0030] Table 1 The name, sequence and position of the ring-mediated is...

Embodiment 2

[0032] Example 2 Design of loop-mediated isothermal amplification primers for detection of type Ⅱ porcine circovirus

[0033]In this embodiment, the loop-mediated isothermal amplification primers for detecting type II porcine circovirus include a first outer primer, a second outer primer, a first inner primer, a second inner primer, and a loop primer; wherein, the first The outer primer (hereinafter referred to as F3) has the sequence structure shown in SEQ ID No.1; the second outer primer (hereinafter referred to as B3) has the sequence structure shown in SEQ ID No.2; the first inner primer The primer (hereinafter referred to as FIP) has the sequence structure shown in SEQ ID No.3; the second inner primer (hereinafter referred to as BIP) has the sequence structure shown in SEQ ID No.4; the loop primer ( Hereinafter referred to as LB) has a sequence structure as shown in SEQ ID No.5. Its specific sequence structure and its position on the ORF1 gene for detecting type II porci...

Embodiment 3

[0036] Example 3 Detection of type Ⅱ porcine circovirus by loop-mediated isothermal amplification primers

[0037] The PK15 cells used in this example were preserved by the Key Laboratory of Animal Immunology, Henan Academy of Agricultural Sciences; the PCV2-ZM strain was clinically isolated by the Key Laboratory of Animal Immunology, Henan Academy of Agricultural Sciences; the DNA extraction kit was purchased from Bioengineering (Shanghai ) Ltd. It should be noted that PK15 cells, PCV2-ZM strains, and DNA extraction kits are all disclosed in the prior art, and their acquisition methods include but are not limited to the methods disclosed in this example, the above-mentioned products obtained from different raw materials, sources, and preparation methods The product does not affect the implementation and effect of the present invention.

[0038] The loop-mediated isothermal amplification primers described in Examples 1 and 2 were used to detect type II porcine circovirus, and...

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Abstract

The invention provides a loop-mediated isothermal amplification primer for detecting type 2 porcine circovirus. The loop-mediated isothermal amplification primer comprises a first outer primer, a second outer primer, a first inner primer and a second inner primer. The invention further provides a method for detecting the type 2 porcine circovirus by the loop-mediated isothermal amplification primer. The loop-mediated isothermal amplification primer can rapidly detect the type 2 porcine circovirus and is high in detection sensitivity, good in specificity, high in detection rate, short in reaction time and simple to operate, and results are conveniently observed.

Description

technical field [0001] The invention belongs to the field of veterinary biomolecular detection, in particular to a ring-mediated isothermal amplification primer and a method for detecting type II porcine circovirus. Background technique [0002] Porcine circovirus (PCV) is one of the smallest animal viruses discovered so far. It is known that PCV has two serotypes, PCV1 and PCV2, wherein PCV1 is a non-pathogenic virus, and PCV2 is a pathogenic virus. PCV2 is the main pathogen of postweaning multisystemic wasting syndrome (PostweaningMultisystemicWastingSyndrome, PMWS), dermatitis and nephrotic syndrome (PNDS). Since Clark first isolated Porcine circovirus type 2 (PCV2) in 1997, the virus has caused huge economic losses to the swine industry all over the world. At the 20th International Swine Disease Conference in 2008, people even listed it as the number one disease in the current pig industry. Therefore, it is necessary to establish a rapid on-site detection method for ty...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/70C12Q1/68C12N15/11
CPCC12Q1/6844C12Q1/6848C12Q1/701C12Q2531/119
Inventor 王德国王永真宋春美肖付刚张永清
Owner XUCHANG UNIV
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