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Rapid detection method and detection kit for porcine ridge virus combined with rt-rpa and lateral flow chromatography

A technology of RT-RPA and detection kit, which is applied in the field of molecular biology detection of viruses, can solve the problems that the detection results cannot be directly observed, the detection equipment is inconvenient to carry, and the reaction procedure is complicated, so as to achieve convenient and fast detection process and high detection results. Safe and reliable, high sensitivity effect

Active Publication Date: 2018-09-28
LANZHOU INST OF VETERINARY SCI CHINESE ACAD OF AGRI SCI
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Problems solved by technology

[0006] In order to solve the problems existing in the known technology, the present invention provides a rapid detection method of porcine crest virus and its detection kit by combining RT-RPA and lateral flow chromatography technology, which can quickly complete porcine crest virus at a lower temperature. The virus detection solves the problems of complex reaction procedures, low sensitivity, inconvenient carrying of detection instruments and inability to directly observe the detection results in the detection of porcine crest virus

Method used

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  • Rapid detection method and detection kit for porcine ridge virus combined with rt-rpa and lateral flow chromatography
  • Rapid detection method and detection kit for porcine ridge virus combined with rt-rpa and lateral flow chromatography
  • Rapid detection method and detection kit for porcine ridge virus combined with rt-rpa and lateral flow chromatography

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Embodiment 1

[0035] The porcine crista virus rapid detection kit combining RT-RPA and lateral flow chromatography technology of the present invention includes;

[0036] RNA extraction reagents, primers, probes, TR-RPA amplification reaction reagents, lateral flow chromatography test strips, negative controls;

[0037] Primer and probe design

[0038] According to the whole genome sequence of porcine kobuvirus published by GenBank, DNA Star software was used for sequence alignment to find out the conserved gene sequence regions, and a pair of primers and a probe were designed using the probe design software Primer Express5.0. The specific information is shown in Table 1 below. The primers and probes were synthesized by Dalian Bao Biological Company.

[0039] Table 1 Porcine crista virus primers and probe sequences

[0040]

[0041] The primer sequences are:

[0042] Upstream primer: Kobu-RPA-7379-F:

[0043] 5'-TTGGCAACGAGACGTATGAGATGATTGGCGGTAA-3';

[0044] Downstream primer: Kobu-...

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Abstract

The invention discloses a specific primer and a probe for quickly detecting porcine kobuvirus by combining reverse transcription-recombinase polymerase amplification (RT-RPA) and a lateral flow chromatographic technology, and creates a porcine kobuvirus quick detection method combining RT-RPA and the lateral flow chromatographic technology. The method can detect 6.4*10<3>copies / mu L of porcine kobuvirus RNA at least, so that the sensitivity is much higher than that of an existing PCR detection method. The method can detect the porcine kobuvirus as early as 10 minutes later the reaction startsand can finish detection within 10-15 minutes. The specific primer and probe designed for quick detection of the porcine kobuvirus have strong specificity for porcine kobuvirus, the detection result can be seen intuitively by naked eyes, the detection process is convenient and quick, and the detection result is safe and reliable.

Description

technical field [0001] The invention belongs to the technical field of molecular biology detection of viruses, and in particular relates to a rapid detection method for porcine crest virus and a detection kit thereof combining RT-RPA and lateral flow chromatography technology. Background technique [0002] In recent years, nucleic acid isothermal amplification technology has developed rapidly. It gets rid of the traditional temperature cycler and can rapidly carry out the amplification reaction in a short time. In 2006, Piepenburg et al. first proposed recombinase polymerase amplification (RPA), which is a new nucleic acid isothermal amplification technology that simulates the DNA replication process in organisms. The amplification process can be performed in traditional reaction tubes. The target fragment can be amplified isothermally at 37°C-42°C, and the amplification reaction can be completed in 20 minutes. Because of its sensitive characteristics, it is widely used in ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/70C12Q1/6844
Inventor 刘新生张永光王永录方玉珍周鹏刘占旭
Owner LANZHOU INST OF VETERINARY SCI CHINESE ACAD OF AGRI SCI
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