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System for simultaneously detecting six pig viruses and special LAMP primer for system

A technology of swine fever virus and porcine foot-and-mouth disease virus, applied in the field of the system and its special LAMP primers, can solve the problems of difficult to make accurate judgment, loss of pig industry, misjudgment and misjudgment, etc.

Active Publication Date: 2017-08-11
BOAO BIOLOGICAL CO LTD +1
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  • Summary
  • Abstract
  • Description
  • Claims
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AI Technical Summary

Problems solved by technology

[0002] Porcine foot-and-mouth disease virus (Foot-and-mouth disease virus, FMDV), classical swine fever virus (CSFV), porcine reproductive and respiratory syndrome virus (Porcine reproductive and respiratory syndrome virus, PRRSV), porcine circovirus type Ⅱ ( Porcine circovirus Ⅱ, PCV-2), porcine parvovirus (PPV), porcine pseudorabies virus (Porcine pseudorabies virus, PRV), the diseases caused by these viruses often present similar symptoms of respiratory and reproductive disorders clinically , and often in mixed infection, causing heavy losses to the pig industry
However, it is difficult to make accurate judgments with conventional etiological detection and clinical pathological changes, especially when multiple pathogens are mixed with infection, it is difficult to carry out early and rapid detection, which is prone to misjudgment and misjudgment

Method used

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  • System for simultaneously detecting six pig viruses and special LAMP primer for system
  • System for simultaneously detecting six pig viruses and special LAMP primer for system
  • System for simultaneously detecting six pig viruses and special LAMP primer for system

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0234] Embodiment 1, complete set of primers

[0235] LAMP primer set for detection of 6 porcine viruses, consisting of 6 primer sets packaged individually. The sequences of each primer in the 6 primer sets are shown in Table 2.

[0236] Table 2

[0237]

[0238]

[0239] Y stands for T or C.

Embodiment 2

[0240] Embodiment 2, preparation detection device

[0241] A microfluidic chip with 24 reaction pools was taken, and agarose aqueous solution was used as an embedding agent, and the primers were embedded and dried.

[0242] The primers embedded in the 24 reaction pools of the microfluidic chip are as follows:

[0243] (1) Blank;

[0244] (2) negative control;

[0245] (3) negative control;

[0246] (4) A primer set for detecting porcine circovirus type Ⅱ;

[0247] (5) A primer set for detecting porcine circovirus type Ⅱ;

[0248] (6) negative control;

[0249] (7) primer set for detecting porcine foot-and-mouth disease virus O / A / Asia I type;

[0250] (8) primer set for detecting porcine foot-and-mouth disease virus O / A / Asia I type;

[0251] (9) negative control;

[0252] (10) a primer set for detecting porcine parvovirus;

[0253] (11) a primer set for detecting porcine parvovirus;

[0254] (12) negative control;

[0255] (13) a primer set for detecting porcine pseu...

Embodiment 3

[0268] Example 3, Detection of Nucleic Acid Samples Using Constant Temperature Amplification Microfluidic Chips

[0269] The viruses to be tested are: porcine foot-and-mouth disease virus O / A / Asia I type, classical swine fever virus, porcine reproductive and respiratory syndrome virus American type, porcine circovirus type II, porcine parvovirus or porcine pseudorabies virus. For details, see Table 1.

[0270] 1. Extract the nucleic acid of the virus to be tested to obtain a nucleic acid concentration of 5×10 3 copies / μl of template solution.

[0271] 2. Mix 9 microliters of the template solution with 16 microliters of the pre-amplification system to obtain a sample injection solution.

[0272] Pre-amplification system (16 μl): containing 12.5 μl of 2×reaction solution (RM), 1.5 μl of enzyme mixture (EM), 19 μM calcein and 0.8 μM manganese chloride.

[0273] The above pre-amplification system is a loop-mediated isothermal amplification pre-amplification system.

[0274] 3....

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Abstract

The invention discloses a system for simultaneously detecting six pig viruses and a special LAMP primer for the system. The system comprises an LAMP complete primer for detecting the six pig viruses and / or a reagent required for LAMP and / or an instrument and / or an amplified product data processor, wherein the amplified product data processor is used for distinguishing whether a to-be-detected sample amplification product obtained through loop-mediated isothermal amplification on a to-be-detected sample through the LAMP complete primer for detecting the six pig viruses comprises a specific amplified product or not. The system for detecting the six pig viruses and the special LAMP primer for the system can meet simultaneous and rapid detection of a pig foot-and-mouth disease virus O / A / Asia type I, a hog cholera virus, a porcine respiratory and reproductive syndrome virus American type, a porcine circovirus type II, a porcine parvovirus and a porcine pseudorabies virus, the detection system is simple in operation, the result is convenient to observe and a strong technical support is provided for epidemic disease monitoring of a pig farm.

Description

technical field [0001] The invention belongs to the technical field of nucleic acid amplification, and relates to a system for simultaneously detecting six kinds of porcine viruses and special LAMP primers thereof. Background technique [0002] Porcine foot-and-mouth disease virus (Foot-and-mouth disease virus, FMDV), classical swine fever virus (CSFV), porcine reproductive and respiratory syndrome virus (Porcine reproductive and respiratory syndrome virus, PRRSV), porcine circovirus type Ⅱ ( Porcine circovirus Ⅱ, PCV-2), porcine parvovirus (PPV), porcine pseudorabies virus (Porcine pseudorabies virus, PRV), the diseases caused by these viruses often present similar symptoms of respiratory and reproductive disorders clinically , and often in mixed infection, causing heavy losses to the pig industry. However, it is difficult to make accurate judgments with conventional etiological detection and clinical pathological changes, especially when multiple pathogens are mixed with ...

Claims

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Application Information

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IPC IPC(8): C12Q1/70C12Q1/68C12N15/11
CPCC12Q1/6844C12Q1/701C12Q2600/16C12Q2531/119C12Q2537/143
Inventor 许俊泉胥慧袁向芬范晓攀吴绍强徐洪涛孙晶华邓企飞王彩霞欧阳兆槐
Owner BOAO BIOLOGICAL CO LTD
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