Duck hepatitis virus type I LAMP (loop-mediated isothermal amplification) detection kit

A technology for detecting kits and hepatitis viruses, which is applied in the determination/inspection of microorganisms, microorganisms, biochemical equipment and methods, etc. It can solve the problems of low sensitivity, time-consuming, difficult standardization, etc., and achieve high sensitivity and simple operation Effect

Active Publication Date: 2012-02-08
GUANGXI VETERINARY RES INST
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] The traditional detection methods of duck hepatitis I virus include serum neutralization test, agar diffusion test and ELISA, etc., but these tests have disadvantages such as time-consuming, low sensitivity, and difficult standardization, which have certain limitations in practical application
At present, although duck hepatitis I virus PCR detection methods and fluorescent

Method used

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  • Duck hepatitis virus type I LAMP (loop-mediated isothermal amplification) detection kit
  • Duck hepatitis virus type I LAMP (loop-mediated isothermal amplification) detection kit
  • Duck hepatitis virus type I LAMP (loop-mediated isothermal amplification) detection kit

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Embodiment Construction

[0023] 1. Preparation of Duck Hepatitis I Virus LAMP Detection Kit

[0024] Reaction solution A: 24 μL per tube, its composition is: 2.5ul 10× isothermal reaction buffer solution, 1ul 5U / ul AMV, 1ul40U / ul inhibitor, 1.0ul Bst DNA polymerase 8U / ul, 3.5ul 10mM dNTPs, 5ul 25mM sulfuric acid Magnesium, 0.25ul 20uM Inner Primer 1, 0.25ul 20uM Inner Primer 2, 2ul 20uM Outer Primer 1, 2ul 20uM Outer Primer 2, 1ul 20uM Loop Primer 1, 1ul 20uM Loop Primer 2, 0.5ul 5M Beet Base, 1ul 625μM calcein and 1ul 12.5mM manganese chloride and 1ul double distilled water.

[0025] Among them, the 10× isothermal reaction buffer solution contains 200mM trishydrochloride with a pH value of 8.8, 100mM potassium chloride, 100mM ammonium sulfate, 20mM magnesium sulfate and 1% Triton X-100.

[0026] The sequence of the inner primer 1 is CWTGTGGCACAGCTCCAAAGGGGTTTTAGTGTTGTGGGA (see sequence listing SEQ.ID.No.2),

[0027] The sequence of the inner primer 2 is CTTGACAGTGCTGTYAGAAACTGCAACTAATACCTAAACAATCAT...

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Abstract

The invention discloses a duck hepatitis virus type I LAMP (loop-mediated isothermal amplification) detection kit. A LAMP technology is adopted; and two specific inner primers, two specific outer primers and two specific ring primers are designed according to a gene conserved region shared by duck hepatitis viruses type I. Due to the application of the duck hepatitis virus type I LAMP detection kit and a detection method established by the invention, the defects of long detection time, large workload, cross pollution, complex operation and the like in the prior art are overcome. The duck hepatitis virus type I LAMP detection kit has the strong specificity and high sensitivity, is rapid, has low cost and simpler operation method and is particularly suitable for the requirement on field rapid detection of the duck hepatitis virus type I in the primary-level areas.

Description

technical field [0001] The invention relates to the field of duck type I hepatitis virus detection, in particular to a duck type I hepatitis virus LAMP detection kit. Background technique [0002] Duck Hepatitis Virus type I is the causative agent of a highly lethal viral infectious disease targeting ducklings. The disease mainly affects ducklings within 4 weeks of age, and the mortality rate is as high as more than 90%. It is one of the most serious infectious diseases that endanger the duck industry. [0003] The traditional detection methods of duck hepatitis I virus include serum neutralization test, agar diffusion test and ELISA, etc., but these tests have disadvantages such as time-consuming, low sensitivity, and difficult standardization, which have certain limitations in practical application. At present, although duck hepatitis I virus PCR detection methods and fluorescent quantitative PCR detection methods have been established, their detection sensitivity is rela...

Claims

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Application Information

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IPC IPC(8): C12Q1/70C12Q1/68C12R1/93
Inventor 谢芝勋谢丽基刘加波谢志勤庞耀珊邓显文范晴
Owner GUANGXI VETERINARY RES INST
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