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Koi herpesvirus RPA primer pair, probe and detection kit and application thereof

A koi herpes virus, detection kit technology, applied in biochemical equipment and methods, microbial determination/inspection, DNA/RNA fragments, etc. Carp herpes virus detection method, high lethality problems, to achieve stable detection effect, shorten detection time, and high sensitivity

Active Publication Date: 2021-06-29
PEARL RIVER FISHERY RES INST CHINESE ACAD OF FISHERY SCI
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Problems solved by technology

But the koi herpes virus seriously affects the normal breeding of carp and koi. It is highly contagious and has a high lethal rate. The fish will die after 24 to 48 hours of onset of symptoms. the harm of
[0004] In related technologies, there is no rapid, sensitive and accurate quantitative detection method for koi herpes virus, nor is there any specific veterinary drug or vaccine for koi herpes virus disease

Method used

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  • Koi herpesvirus RPA primer pair, probe and detection kit and application thereof
  • Koi herpesvirus RPA primer pair, probe and detection kit and application thereof
  • Koi herpesvirus RPA primer pair, probe and detection kit and application thereof

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Embodiment Construction

[0056] In order to make the object, technical solution and technical effect of the present invention clearer, the present invention will be further described in detail below in conjunction with specific embodiments. It should be understood that the specific implementations described in this specification are only for explaining the present invention, not for limiting the present invention.

[0057] The experimental materials and reagents used, unless otherwise specified, are conventional consumables and reagents that can be obtained from commercial channels.

[0058] Primer design and screening

[0059] 1. Acquisition of virus strain DNA:

[0060] The virus strain used in this example is the KHV-GZ1301 strain (preserved in the Pearl River Fisheries Research Institute of the Chinese Academy of Fishery Sciences).

[0061] The KHV-GZ1301 strain was inoculated into the CCB sensitive cell line, and when the cells showed obvious cytopathic changes, the DNA of the virus strain was ...

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Abstract

The invention discloses a koi herpesvirus RPA primer pair, a probe and a detection kit and application thereof. Sequences of the RPA primer pair are as shown in SEQ ID NO.4-19, and the sequences of the probe are as shown in SEQ ID NO.2-3. The probe and the primers provided by the invention are high in sensitivity, stable in detection effect and high in specificity. On the basis, the constructed detection reagents and detection kit have very high detection sensitivity, the lowest detection limit is 100 copy / mu L, and the detection time is only 4-18 minutes. Compared with KHV detection gold standard PCR and real-time PCR, the detection reagents and the detection kit have the advantages of time and detection accuracy, with extremely high application value.

Description

technical field [0001] The invention belongs to the technical field of gene detection, and in particular relates to a koi herpes virus RPA primer set, a probe, a detection kit and applications thereof. Background technique [0002] Koi herpesvirus belongs to the order Herpesvirales, the family Alloherpesviridae, and the genus Cyprinivirus. It is a double-stranded DNA virus with an envelope. The diameter of the virus particle is 167-200nm. 295kbp, encoding a total of 156 open reading frames (Open reading frame, ORF), is the largest known herpes virus genome. Koi herpesvirus (Koi herpesvirus, KHV), also known as cyprinid herpesvirus 3 (CyHV-3), is a disease that causes carp (Cyprinus carpio), koi (Cyprinus carpio koi) and its variants to infect koi The pathogen of herpes virus disease (Koi herpesvirus disease, KHVD). Koi herpes virus disease has been widely distributed all over the world. The World Organization for Animal Health has listed koi herpes virus as a fish infectio...

Claims

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Application Information

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IPC IPC(8): C12Q1/70C12Q1/6844C12N15/11C12R1/93
CPCC12Q1/6844C12Q1/705C12Q2600/136C12Q2521/107C12Q2521/507C12Q2522/101C12Q2531/119C12Q2563/107
Inventor 李莹莹王庆尹纪元王英英吴斯宇曾伟伟常藕琴任燕石存斌
Owner PEARL RIVER FISHERY RES INST CHINESE ACAD OF FISHERY SCI
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