H6 subtype avian influenza virus RT-LAMP (reverse transcription loop-mediated isothermal amplification) kit

A technology of RT-LAMP and avian influenza virus, which is applied in the field of H6 subtype avian influenza virus RT-LAMP kit, can solve the problems of long detection cycle, low specificity, and low sensitivity, and achieve the effect of short reaction time

Active Publication Date: 2012-02-08
GUANGXI VETERINARY RES INST
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  • Abstract
  • Description
  • Claims
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AI Technical Summary

Problems solved by technology

However, these methods have long detection cycle, low sensitivity, low specificity, and are not suitable for grass-roots live poultry markets or the detection of a large number of samples imported and exported.

Method used

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  • H6 subtype avian influenza virus RT-LAMP (reverse transcription loop-mediated isothermal amplification) kit
  • H6 subtype avian influenza virus RT-LAMP (reverse transcription loop-mediated isothermal amplification) kit
  • H6 subtype avian influenza virus RT-LAMP (reverse transcription loop-mediated isothermal amplification) kit

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Embodiment Construction

[0028] In the following, the H6 subtype avian influenza virus RT-LAMP kit of the present invention will be further described in detail in conjunction with the accompanying drawings and specific embodiments.

[0029] 1. Preparation of H6 subtype avian influenza virus RT-LAMP kit

[0030] Reaction Solution A

[0031] 23 μL per tube, which consists of: 2.5 μL 10× isothermal reaction buffer, 1.0 μL AMV 5U / μL, 1.0 μL Rnasin 20 U / μL, 1.0 μL Bst DNA PoLymerase 8 U / μL, 5 μL 25 mM magnesium sulfate, 3.5 μL 10 mM dNTP, 5 μL 5M betaine, 0.5 μL of 10 μM outer primer 1 and 0.5 μL of 10 μM outer primer 2, 1.0 μL of 30 μM inner primer 1 and 1.0 μL of 30 μM inner primer 2, 0.5 μL of 40 μM loop primer 1 and 0.5 μL of 40 μM loop primer 2.

[0032] Wherein, the 10× isothermal reaction buffer contains 200 mM trishydrochloride with a pH value of 8.8, 100 mM potassium chloride, 100 mM ammonium sulfate, 20 mM magnesium sulfate and 1% Triton X-100.

[0033] The sequence of the outer primer 1 is GG...

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Abstract

The invention discloses an H6 subtype avian influenza virus RT-LAMP (reverse transcription loop-mediated isothermal amplification) kit. A reverse transcription loop-mediated isothermal amplification (RT-LAMP) technology is adopted; and six specific primers of LAMP are designed according to eight loci in a conserved region of an H6 subtype AIV (avian influenza virus) hemagglutinin (HA) gene sequence. Due to the application of the H6 subtype avian influenza virus RT-LAMP kit and a detection method established by the invention, the defects of long detection period, low sensitivity, poor specificity, complex operation, high instrument requirement in the prior art are overcome. The H6 subtype avian influenza virus RT-LAMP kit is particularly suitable for the requirement on field detection of the H6 subtype avian influenza virus in the primary-level areas and frontier ports and has important significance for preventing and controlling the H6 subtype avian influenza virus.

Description

technical field [0001] The invention relates to the field of bird flu virus detection, in particular to an H6 subtype bird flu virus RT-LAMP kit. Background technique [0002] Avian influenza AI is an acute and severe infectious disease listed as Class A by the Organization for Animal Health (OIE), and it is also listed as a Class I disease in China. AI is caused by the avian influenza virus AIV, which mainly causes respiratory diseases in poultry and is characterized by acute onset, high mortality and decreased egg production rate. Although H6 subtype AIV (H6 subtype avian influenza virus) is not a highly pathogenic avian influenza virus, it can be transmitted among poultry, causing invisible infection of poultry, and may also drift or transform into a new, more virulent virus strain, so early, rapid and accurate diagnosis is very important. [0003] Conventional avian influenza virus diagnosis methods include: virus isolation, hemagglutination (HA) and hemagglutination i...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/70C12Q1/68C12R1/93
Inventor 谢芝勋周辰瑜刘加波庞耀珊邓显文谢志勤谢丽基
Owner GUANGXI VETERINARY RES INST
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