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Porcine circovirus type 2 rapid typing detection kit

A detection kit, porcine circovirus technology, applied in the field of porcine circovirus type 2 rapid typing detection kit, can solve the problems such as insufficient primer specificity

Active Publication Date: 2013-07-31
湖南派智生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, in the detection system of the PCV2 real-time quantitative PCR reaction system, due to the lack of specificity of the primers, probes are needed to ensure the specificity of the reaction

Method used

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  • Porcine circovirus type 2 rapid typing detection kit
  • Porcine circovirus type 2 rapid typing detection kit
  • Porcine circovirus type 2 rapid typing detection kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0016] Example 1, quantitative and qualitative PCV in real-time SMART fluorescence.

[0017] Step 1: In PCR real-time reaction tube a, add 10 μl 2X SMART Master Mix (including EvaGreen fluorescent dye, SMART buffer system, SMART Taq, dNTP, magnesium ions), 500 nM PCV2a forward primer and PCV2a reverse primer, and 1 μl To measure nucleic acid samples, add the total reaction volume to 20 μl with pure water.

[0018]Step 2: Add 10 μl 2X SMART Master Mix, 500 nM PCV2b forward primer and PCV2b reverse primer, and 1 μl nucleic acid sample to be tested in PCR real-time reaction tube b, and bring the total reaction volume to 20 μl with pure water.

[0019] Step 3: Add 10 μl 2X SMART Master Mix, 500 nM PCV2c forward primer and PCV2c reverse primer, and 1 μl nucleic acid sample to be tested in PCR real-time reaction tube c, and bring the total reaction volume to 20 μl with pure water.

[0020] Step 4: Place the above three tubes in ABI7500 (or ABI7900, or BioRad, Bioer, Roche, instrume...

Embodiment 2

[0024] Example 2, SMART has higher specificity in real-time fluorescent quantitative PCR method

[0025] The present invention compares the specificity of SMART PCR reaction and ordinary PCR represented by SYBR GreenER. The result is as figure 1 shown. figure 1 The upper panel shows that 10 μL SMART PCR reaction system contains 250 nM positive and negative primers for detecting PCV2a (shown in Table 1), and one million copies of PCV2a, or PCV2b, or PCV2c, or NTC . figure 1 The lower panel shows 10 μL of SYBR GreenER as a representative of the common reaction system containing 250 nM concentration of positive and negative primers for detecting PCV2a (shown in Table 1), and one million copies of PCV2a, or PCV2b, or It is PCV2c, or NTC.

[0026] Table 1

[0027]

[0028] figure 1 It clearly shows that the SMART PCR reaction is specific, and no cross-reaction is found, while the common reaction system represented by SYBR GreenER has cross-reaction with the PCV2b template....

Embodiment 3

[0029] Example 3, SMART has higher specificity in real-time fluorescence quantitative PCR method

[0030] The present inventors again compared the specificity of SMART PCR reaction and ordinary PCR represented by SYBR GreenER. The result is as figure 2 shown. figure 2 The upper panel shows that 10 μL of the SMART PCR reaction system contains 250 nM concentration, used to detect the positive and negative primers of PCV2b (shown in Table 1), and one million copies of PCV2a, or PCV2b, or PCV2c, respectively, Or NTC. figure 2 The lower panel shows that the common reaction system represented by 10μLSYBR GreenER contains 250nM positive and negative primers for detecting PCV2b (shown in Table 1), and one million copies of PCV2a, or PCV2b, or PCV2c respectively , or NTC.

[0031] figure 2 It clearly shows that the SMART PCR reaction and specificity have no cross-reaction, while the common reaction system represented by SYBR GreenER has cross-reaction with PCV2a template and P...

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Abstract

The invention relates to a porcine circovirus type 2 rapid typing detection kit which belongs to the field of biological detection and is a kit based on a SMART detection system method. By the adoption of the kit, the technical problem that a porcine circovirus type 2 (PCV2a, 2b, 2d, etc.) typing detection method requires a complex operational program, high experiment skill, long experimental period and the like is solved. A rapid and accurate molecular diagnosis technical product can be provided for porcine circovirus type 2 typing. The kit provided by the invention can be used in system diagnosis of porcine circovirus type 2 rapid typing and used to guide prevention and treatment of the disease caused by porcine circovirus type 2. And mutation detection of each type sequence of the virus can be realized, and the kit is beneficial to molecular epidemiological investigation of the porcine circovirus, meat product quarantine inspection, viral surveillance of forage and water quality in a pig farm and the like.

Description

technical field [0001] The invention relates to a porcine circovirus type 2 rapid typing detection kit, which belongs to the field of biological detection. Background technique [0002] Porcine circovirus (porcine circovirus type 2, PCV-2) belongs to the family Circoviridae, with a diameter of only 17nm, no envelope, icosahedral symmetry, covalently closed circular single-stranded DNA virus. PCV-2 is the main pathogen that causes postweaning multisystemic wasting syndrome (PMWS) in piglets. Since the first outbreak of the disease in Canadian pigs in 1991, it has been widely prevalent in pigs all over the world. The pig industry has caused huge economic losses. PCV-2 is often associated with porcine reproductive and respiratory syndrome virus (porcine reproductive and respiratory syndrome virus, PRRSV), porcine parvovirus (porcine parvovirus, PPV), porcine pseudorabies virus (pseudorabies virus, PRV), Haemophilus parasuis ( Haemophilus parasuis (HPs), Actinobacillus pleurop...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/70C12Q1/68C12R1/93
Inventor 杨毅忻星王乃东邓治邦薛立群湛洋首易君
Owner 湖南派智生物科技有限公司
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