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Goose parvovirus detection kit and method based on loop-mediated isothermal amplification technology

A goose parvovirus, ring-mediated isothermal technology, applied in the field of biological detection kits, to achieve the effect of strong specificity, low detection cost, simple and fast operation

Inactive Publication Date: 2010-10-27
CHONGQING ACAD OF ANIMAL SCI +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

But so far there is no report on the detection of goose parvovirus based on the loop-mediated isothermal amplification technique

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] 1. Goose parvovirus detection kit based on loop-mediated isothermal amplification technology, consisting of the following reagents:

[0033] a. The upstream internal primer FIP aqueous solution and the downstream internal primer BIP aqueous solution with a concentration of 5 μmol / L, the upstream external primer F3 aqueous solution and the downstream external primer B3 aqueous solution with a concentration of 20 μmol / L: the primer sequences are as follows:

[0034] Upstream inner primer FIP: 5'-tcgcaatgccaatttcccgagg-ggagctatgggcgactct-3';

[0035] Downstream inner primer BIP: 5'-gaccaccagaacctgggtcc-gcatcttgagaggttccgc-3';

[0036] Upstream outer primer F3: 5'-aatggcagaggggaggagg-3';

[0037] Downstream outer primer B3: 5'-cccaggggggtactgtatcc-3';

[0038] b. An aqueous solution of Bst DNA polymerase with a concentration of 8 U / μl;

[0039] c, 10× thermal polymerization reaction buffer: Tris-HCl with a concentration of 250mmol / L and a pH of 8.8, potassium chloride wi...

Embodiment 2

[0050] 1. The goose parvovirus detection kit based on the loop-mediated isothermal amplification technology is different from the kit described in Example 1 in that the specific primer sequences are different. The primer sequences of this kit are as follows:

[0051] Upstream inner primer FIP: 5'-ggccaaatcctccgagattcgg-cagggacctattggggca-3';

[0052] Downstream inner primer BIP: 5'-caatccaccaccgcaggtgt-ccacttctggtgcacgtatt-3';

[0053] Upstream outer primer F3: 5'-ggtttggcagaacagggata-3';

[0054] Downstream outer primer B3: 5'-gcccgtagagtactgggtta-3'.

[0055] 2. The method for detecting goose parvovirus using the goose parvovirus detection kit based on the loop-mediated isothermal amplification technology is the same as the method described in Example 1. It was found that the color of the blank control group was yellow, indicating that it did not contain Goose parvovirus; the color of the experimental group changed to green, indicating that it contained goose parvovirus, w...

Embodiment 3

[0057] 1. The goose parvovirus detection kit based on the loop-mediated isothermal amplification technology is different from the kit described in Example 1 in that the specific primer sequences are different. The primer sequences of this kit are as follows:

[0058] Upstream inner primer FIP: 5'-ggccaaatcctccgagattcgg-tggggcaaaaataccgaaga-3';

[0059] Downstream inner primer BIP: 5'-caatccaccaccgcaggtgt-ccacttctggtgcacgtatt-3';

[0060] Upstream outer primer F3: 5'-ggtttggcagaacagggata-3';

[0061] Downstream outer primer B3: 5'-gcccgtagagtactgggtta-3'.

[0062] 2. The method for detecting goose parvovirus using the goose parvovirus detection kit based on the loop-mediated isothermal amplification technology is the same as the method described in Example 1. It was found that the color of the blank control group was yellow, indicating that it did not contain Goose parvovirus; the color of the experimental group changed to green, indicating that it contained goose parvovirus,...

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PUM

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Abstract

The invention discloses goose parvovirus detection kit and method based on a loop-mediated isothermal amplification technology. Based on six specific regions of a VP3 gene conserved region of a goose parvovirus, two specific primers and two specific outer primers are designed in the kit so as to ensure the high specificity and the reliability of a detection result of loop-mediated isothermal amplification. The invention detects the goose parvovirus on the basis of the loop-mediated isothermal amplification technology, can amplify a target sequence rapidly, efficiently and specifically under the isothermal condition, has simple and convenient operation and does not use expensive instruments and reagents; an amplification product can be developed directly by using a fluorescent dye and a result can be judged with naked eyes; the detection cost is low; and the invention is particularly suitable for small and medium size units and field tests.

Description

technical field [0001] The invention relates to a biological detection kit and method, in particular to a goose parvovirus detection kit and method based on loop-mediated isothermal amplification technology. Background technique [0002] Goose Parvovirus (GPV) is a non-enveloped single-stranded DNA virus belonging to the Parvoviridae family (Parvoviridate) and the genus Parvovirus (Parvovirus Genus). , also known as gosling plague. The disease is a highly contagious infectious disease with high morbidity and mortality, which has become prevalent in the world, causing serious harm and great economic losses to the goose industry. [0003] At present, the detection methods of goose parvovirus mainly include isolation and culture identification, polymerase chain reaction (PCR) and fluorescence quantitative PCR (FQ-PCR). The PCR method requires expensive instruments and reagents, and the detection cost is high, so it is not suitable for small and medium-sized units and on-site ...

Claims

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Application Information

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IPC IPC(8): C12Q1/70C12Q1/68C12R1/93
Inventor 杨金龙杨睿杨松全程安春付利芝杨柳沈克飞
Owner CHONGQING ACAD OF ANIMAL SCI
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