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 parvovir

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Example Embodiment

[0031] Example 1

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

[0033] a. The upstream inner primer FIP aqueous solution and the downstream inner primer BIP aqueous solution each with a concentration of 5 μmol / L, the upstream outer primer F3 aqueous solution and the downstream outer primer B3 aqueous solution with a concentration of 20 μmol / L each: the primer sequence is 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’-aatggcagagggaggagg-3’;

[0037] Downstream outer primer B3: 5’-cccagggggtactgtatcc-3’;

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

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

Example Embodiment

[0049] Example 2

[0050] 1. The goose parvovirus detection kit based on loop-mediated isothermal amplification technology is different from the kit described in Example 1 in that the specific primer sequence is different. The primer sequence of this kit is 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 loop-mediated isothermal amplification technology-based goose parvovirus detection kit is the same as the method described in Example 1. As a result, the color of the blank control group is yellow, indicating that it does not contain Goose parvovirus; the color of the experimental group changes to green, indicating that it contains goose parvovirus, which is consis...

Example Embodiment

[0056] Example 3

[0057] 1. The goose parvovirus detection kit based on loop-mediated isothermal amplification technology is different from the kit described in Example 1 in that the specific primer sequence is different. The primer sequence of this kit is 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 loop-mediated isothermal amplification technology-based goose parvovirus detection kit is the same as the method described in Example 1. As a result, the color of the blank control group is yellow, indicating that it does not contain Goose parvovirus; the color of the experimental group changes to green, indicating that it contains goose parvovirus, which is cons...

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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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