LAMP (loop-mediated isothermal amplification) kit for detecting acute hepatopancreatic necrosis disease pathogen

A kit, hepatopancreas technology, applied in the field of microbial detection, to achieve the effect of high sensitivity, convenient operation and good specificity

Active Publication Date: 2017-12-29
广州双螺旋基因技术有限公司 +1
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

At present, there is no application of LAMP technology in the prior art to the pathogen of acute hepatopancreatic necrosis AP2 genetic testing

Method used

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  • LAMP (loop-mediated isothermal amplification) kit for detecting acute hepatopancreatic necrosis disease pathogen
  • LAMP (loop-mediated isothermal amplification) kit for detecting acute hepatopancreatic necrosis disease pathogen
  • LAMP (loop-mediated isothermal amplification) kit for detecting acute hepatopancreatic necrosis disease pathogen

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Embodiment 1

[0041]A LAMP kit for detecting the pathogen of acute hepatopancreatic necrosis, consisting of: a set of LAMP primers for detecting the pathogen of acute hepatopancreatic necrosis, DNA polymerase, 2× reaction buffer, fluorescent dye, sealing solution, positive standard Template and negative controls.

[0042] The LAMP primer set for detecting the pathogen of acute hepatopancreatic necrosis consists of outer primers, inner primers and loop primers, uses the AP2 gene of the pathogenic plasmid PAV1 of acute hepatopancreatic necrosis as the target gene, and uses the primer design software PrimerExplorer 4.0 to design the following: The 6 LAMP primers including the loop primers were synthesized by Sangon Bioengineering (Shanghai) Co., Ltd. The sequences of the outer primers are shown in SEQ ID NO:1~SEQ ID NO:2, and the sequences of the inner primers are shown in SEQ ID NO : 3 to SEQ ID NO: 4, the loop primer sequence is shown in SEQ ID NO: 5 to SEQ ID NO: 6.

[0043] The molar rati...

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Abstract

The invention discloses a LAMP (loop-mediated isothermal amplification) primer group for detecting acute hepatopancreatic necrosis disease pathogen. The LAMP primer group comprises an outer primer, an inner primer and a loop primer. The sequence of the outer primer is as shown in SEQ ID NO: 1-SEQ ID NO: 2, the sequence of the inner primer is as shown in SEQ ID NO: 3-SEQ ID NO: 4, and the sequence of the loop primer is as shown in SEQ ID NO: 5-SEQ ID NO: 6. The invention further discloses a LAMP kit for detecting the acute hepatopancreatic necrosis disease pathogen. The kit comprises the LAMP primer group for detecting the acute hepatopancreatic necrosis disease pathogen, DNA (deoxyribonucleic acid) polymerase, 2*reaction buffer, fluorescent dye, sealing liquid, a standard positive template and a negative control. The kit can directly detect the acute hepatopancreatic necrosis disease pathogen from a complex sample, has the advantages of strong specificity, high sensitivity, good experimental repeatability and convenience and rapidness in operation, can finish detection without expensive instruments and can be used for rapidly and accurately detecting the acute hepatopancreatic necrosis disease pathogen on site.

Description

technical field [0001] The invention relates to the technical field of microbial detection, in particular to a LAMP kit for detecting the pathogen of acute hepatopancreatic necrosis. Background technique [0002] Vannamei early mortality syndrome (Early Mortality Syndrome, EMS) is defined as acute hepatopancreatic necrosis (Acute Hepatopancreatic Necrosis disease, AHPND) in Thailand. Since its discovery in 2009, acute hepatopancreatic necrosis disease (AHPND) has broken out worldwide. However, in the early days, this situation was often considered by farmers to be a problem of seed quality and was not taken seriously. The disease generally spreads rapidly 20 to 30 days after the seedlings are thrown, and the mortality rate increases significantly. After the onset, prawns usually show symptoms such as lethargy, anorexia, slow growth, pale color, and soft shell. The mortality rate of prawns caused by acute hepatopancreatic necrosis can be as high as 100%, which is extremely...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12Q1/04C12N15/11C12R1/63
CPCC12Q1/6844C12Q1/689C12Q2531/119C12Q2563/107C12Q2545/113
Inventor 黄燕琼柏建山邓艳谢会张璜高美玲
Owner 广州双螺旋基因技术有限公司
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