Duck enteritis virus detection kit and method based on loop-mediated isothermal amplification technology

A ring-mediated isothermal and duck enteritis virus technology, which is applied in the direction of microorganism-based methods, biochemical equipment and methods, and microorganism measurement/inspection, to achieve the effects of strong specificity, low detection cost, simple and fast operation

Inactive Publication Date: 2013-01-09
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 duck enteritis virus based on the loop-mediated isothermal amplification technique

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] 1. A detection kit for duck enteritis virus based on loop-mediated isothermal amplification technology, which consists of the following reagents:

[0037] 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 each with a concentration of 20 μmol / L: The primer sequences are as follows:

[0038] Upstream inner primer FIP: 5'-tccacgtcagtttgccgttc-atgtttggcattctacattcg-3';

[0039] Downstream inner primer BIP: 5'-cagcgtaccacagataagtattgaa-tcgatattttagcagaagttcc-3';

[0040] Upstream outer primer F3: 5'-ggataccgtctaatggctc-3';

[0041] Downstream outer primer B3: 5'-tcaaaagctgcgtagagc-3';

[0042]b. Bst DNA polymerase aqueous solution with a concentration of 8 U / μl;

[0043] c. 10× thermal polymerization reaction buffer: Tris-HCl with a concentration of 250 mmol / L and pH of 8.8, potassium ...

Embodiment 2

[0054] 1. The duck enteritis virus detection kit based on 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:

[0055] Upstream inner primer FIP: 5'-actcagcgaggaggaggaaa-tctgctaaaatatcgaaagctct-3';

[0056] Downstream inner primer BIP: 5'-cctgggtacaagcgcacttc-ctgtgagagtgacgaaacc-3';

[0057] Upstream outer primer F3: 5'-ccttaatttcgatggggaact-3';

[0058] Downstream outer primer B3: 5'-aaacgctttattccagaaaca-3'.

[0059] 2. The method of using the duck enteritis virus detection kit based on the loop-mediated isothermal amplification technology to detect duck enteritis virus is the same as the method described in Example 1. The results found that the color of the blank control group was yellow, indicating that it did not contain Duck enteritis virus; the color of the experimental group changed to green, indicating that it contained...

Embodiment 3

[0061] 1. The duck enteritis virus detection kit based on 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:

[0062] Upstream inner primer FIP: 5'-gcgtagagctttcgatattttagca-atttattgtaaaatgtgcgacct-3';

[0063] Downstream inner primer BIP: 5'-cgattttcctcctcctcgct-gcagcactgctatcttcg-3';

[0064] Upstream outer primer F3: 5'-cgtaccacagataagtattgaag-3';

[0065] Downstream outer primer B3: 5'-tgtgagagtgacgaaacc-3'.

[0066] 2. The method of using the duck enteritis virus detection kit based on the loop-mediated isothermal amplification technology to detect duck enteritis virus is the same as the method described in Example 1. The results found that the color of the blank control group was yellow, indicating that it did not contain Duck enteritis virus; the color of the experimental group changed to green, indicating that it contai...

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PUM

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Abstract

The invention discloses duck enteritis virus detection kit and method based on a loop-mediated isothermal amplification technology. Based on six specific regions of a gene conserved region of a duck enteritis virus, 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 duck enteritis virus on the basis of the loop-mediated isothermal amplification technology, can amplify a target sequence rapidly, efficiently and specificallyunder 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 dyeand 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 duck enteritis virus detection kit and method based on loop-mediated isothermal amplification technology. Background technique [0002] Duck enteritis virus (DEV), also known as duck plague virus (DPV), mainly infects ducks, geese and a variety of Anseriformes and causes an acute and fatal infectious disease - duck viral enteritis (Duck virus enteritis). Viral enteritis (DVE), commonly known as Duck plague (DP), often causes huge economic losses and seriously affects the healthy development of the waterfowl breeding industry. [0003] At present, the detection methods of duck enteritis virus mainly include isolation and culture identification method, polymerase chain reaction (PCR) method and fluorescence quantitative PCR (FQ-PCR) method, but the isolation and culture identification method is cumbersome and time-consuming. The PCR method requires expensive instruments a...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/70C12Q1/68C12R1/93
Inventor 杨金龙程安春付利芝杨睿杨松全沈克飞杨柳
Owner CHONGQING ACAD OF ANIMAL SCI
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