Giant salamander iridescent virus taqman real-time fluorescent quantitative PCR kit and its application

A giant salamander iridovirus, real-time fluorescence quantitative technology, applied in fluorescence/phosphorescence, determination/inspection of microorganisms, biochemical equipment and methods, etc.
CN102286642AActive Publication Date: 2011-12-21YANGTZE RIVER FISHERIES RES INST CHINESE ACAD OF FISHERY SCI
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Patent Information

Authority / Receiving Office
CN · China
Patent Type
Applications(China)
Current Assignee / Owner
YANGTZE RIVER FISHERIES RES INST CHINESE ACAD OF FISHERY SCI
Publication Date
2011-12-21

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Abstract

The invention discloses a giant salamander iridovirus TaqMan real-time fluorescent quantitative PCR (polymerase chain reaction) kit and application thereof. The kit comprises sterile double distilled water, a 10*Taq reaction buffer, 2.5mmol / L dNTPs (deoxyribonucleotide triphosphates) and a fluorescent quantitative reaction liquid. The kit is characterized in that the fluorescent quantitative reaction liquid contains primers and a fluorescent probe, wherein the primers are a forward primer and a reverse primer, the forward primer is 5'-GCGGTTCTCACACGCAGTC-3', the reverse primer is 5'-ACGGGAGTGACGCAGGTGT-3', and the fluorescent probe sequence is 5'-AGCCGACGGAAGGGTGTGTGAC-3'; the 5' terminal marker of the fluorescent probe is FAM, and the 3' terminal marker is ROX; 5U / mu L Taq DNA (deoxyribonucleic acid) polymerase and standard positive template pMCP are a pMD19-T vector composed of 1392 nucleotide segments containing giant salamander iridovirus MCP gene; and the vector proliferates in colibacillus E.coli DH5a. The invention also discloses application of the kit in detection of giant salamander iridovirus pathogen. The kit is quantitative and accurate, and has the advantage of high detection speed (the detection only takes 1 hour, and totally takes 2-3 hours plus the extraction and preparation of the DNA); the method is easy to implement and simple to operate; and the invention can be used for simultaneously detecting high-flux samples, and the accuracy is up to 98%.
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Description

technical field

[0001] The invention belongs to the technical field of giant salamander disease detection, and more specifically relates to a giant salamander iridescent virus TaqMan real-time fluorescent quantitative PCR kit. The PCR kit can not only use all types of fluorescent quantitative PCR instruments currently on the market, but more importantly, can quickly Quantitative detection of giant salamander iridescent virus. At the same time, it also relates to the use of a giant salamander iridescent virus TaqMan real-time fluorescent quantitative PCR kit. technical background

[0002] The giant salamander (Andrias davidiamus), commonly known as the giant salamander, is a second-class national protected animal and is included in the CITES (Convention on International Trade in Endangered Species of Wild Fauna and Flora) species catalog. It is the largest existing individual amphibian and has important scientific research value. and medicinal and nutritional value. However...

Claims

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