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Dual fluorescent rt-lamp detection group, kit and application for visual identification of foot-and-mouth disease virus and bluetongue virus

A technology of RT-LAMP and bluetongue virus, applied in the field of double fluorescence RT-LAMP detection group, can solve problems such as uncertainty

Active Publication Date: 2020-10-20
GUANGXI VETERINARY RES INST
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although many scholars at home and abroad have established multiple LAMP detection methods, these methods have certain shortcomings: they can only detect whether the sample is diseased, but cannot determine which pathogen caused the result

Method used

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  • Dual fluorescent rt-lamp detection group, kit and application for visual identification of foot-and-mouth disease virus and bluetongue virus
  • Dual fluorescent rt-lamp detection group, kit and application for visual identification of foot-and-mouth disease virus and bluetongue virus
  • Dual fluorescent rt-lamp detection group, kit and application for visual identification of foot-and-mouth disease virus and bluetongue virus

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Experimental program
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Effect test

Embodiment 1

[0029] 1. Materials and Methods

[0030] 1.1 Reagents and equipment

[0031] LAMP RNA amplification kit, Loopamp LA-320C real-time turbidimeter was purchased from Japan Eiken Company; RNA / DNA extraction kit, plasmid mini-extraction kit were purchased from Quanshijin Company; premixTaq PCR kit was purchased from Dalian The pGM-T vector was purchased from Tiangen Company; the T7 in vitro transcription kit was purchased from Ferments; the NanoDrop 2000 nucleic acid analyzer was purchased from ThermoFisher Scientific; the multicolor fluorescence imaging analysis system was purchased from BIO-RAD.

[0032] 1.2 Strains

[0033] Foot-and-mouth disease type O, type A, and Asia Ⅰ inactivated vaccines were purchased from Lanzhou Veterinary Research Institute, foot-and-mouth disease type O, A, Asia Ⅰ type inactivated virus, bluetongue inactivated virus (type 4, 8, 9, 15) , Type 17, Type 18), Peste des petits ruminants inactivated virus (PPRV), vesicular stomatitis (VSV) inactivated vir...

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Abstract

The invention discloses a duplex fluorescent RT-LAMP detection group and kit for visually identifying foot-and-mouth disease virus and Bluetongue virus and an application of the detection group and the kit. The duplex fluorescent RT-LAMP detection group comprises two groups of specific primers and probes, the first group of specific primers and probes comprise FMDV-F3, FMDV-B3, FMDV-FIB(F1c-F2), FMDV-BIP(B1c-B2) and FMDV-probes, the second group of specific primers and probes comprise BTV-F3, BTV-B3, BTV-FIB(F1c-F2), BTV-BIP(B1c-B2) and BTV-probes, and the two groups of specific primers and probes are as shown in a sequence table SEQ ID No.1 to SEQ ID No.10. The duplex fluorescent RT-LAMP detection group can identify and diagnose the foot-and-mouth disease virus and the Bluetongue virus inthe same reaction tube and has the advantages of good specificity, high sensibility, less pollution, convenience, rapidness and the like, a detection result can be directly observed by naked eyes andis judged according to colors of reaction products, and the detection group can be used for basic-level quarantine with poor conditions.

Description

technical field [0001] The invention relates to the technical field of bovine virus detection, in particular to a dual fluorescent RT-LAMP detection group, kit and application thereof for visually distinguishing foot-and-mouth disease virus and bluetongue virus. Background technique [0002] Foot-and-mouth disease is an acute and highly contagious infectious disease common to cloven-hoofed animals caused by foot-and-mouth disease virus (FMDV). There are more than 70 kinds of domestic and wild mammals in 20 families, including sheep, camels and sheep. Bluetongue disease is a non-contact infectious disease of ruminants caused by bluetongue virus (BTV). It can infect hosts through insects such as Culicoides, mainly infecting sheep, cattle and wild ruminants. Animals infected with the two viruses showed elevated body temperature, depression, and blisters and ulcers on the oral mucosa, teats and hoofs. The symptoms were similar and difficult to distinguish. It is listed as a le...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/70C12Q1/6844C12N15/11
CPCC12Q1/6844C12Q1/701C12Q2531/119C12Q2537/143C12Q2563/107C12Q2521/107
Inventor 谢芝勋范晴谢志勤谢丽基黄娇玲熊文婕张艳芳曾婷婷王盛罗思思
Owner GUANGXI VETERINARY RES INST
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