Bluetongue virus-8 detection reagent, detection method and application
A bluetongue virus and reagent technology, applied in biochemical equipment and methods, microbial determination/inspection, DNA/RNA fragments, etc., can solve problems such as difficulty in applying on-site detection, lack of cross-immune protection, and complex detection process, etc. Achieve the effect of preventing the spread of the epidemic, facilitating on-site testing, and ensuring production safety
A bluetongue virus and reagent technology, applied in biochemical equipment and methods, microbial determination/inspection, DNA/RNA fragments, etc., can solve problems such as difficulty in applying on-site detection, lack of cross-immune protection, and complex detection process, etc. Achieve the effect of preventing the spread of the epidemic, facilitating on-site testing, and ensuring production safety
CN110257560AActive Publication Date: 2019-09-20SHENZHEN CUSTOMS ANIMAL & PLANT INSPECTION & QUARANTINE TECH CENT +1
Image
Smart Image Click on the blue labels to locate them in the text.
Smart ImageViewing Examples
Examples
Experimental program
Comparison scheme
Effect test
Embodiment
[0040] 1. Materials and methods
[0041] 1. Plasmid and nucleic acid samples for testing
[0042] In this case, according to the bluetongue virus type 8 VP2 gene sequence published in NCBI GenBank, the gene fragment was synthesized by Sangon Bioengineering (Shanghai) Co., Ltd. and cloned into pUC vector, named BTV8-VP2. The inactivated antigenic nucleic acids used in this example include 24 bluetongue virus nucleic acids, deer epidemic hemorrhagic fever virus nucleic acids, bovine viral diarrhea virus nucleic acids and foot-and-mouth disease virus nucleic acids. Among them, twenty-four bluetongue virus nucleic acids are BTV-1 to BTV-24.
[0043] 2. Main reagents and instruments
[0044] TwistAmp exo kit (TwistDx), MMLV Reverse Transcriptase (Takara), Recombinant RNase Inhibitor (Takara), constant temperature amplification instrument (Axxin), 5415R high-speed centrifuge (Eppendorf), 7500 Fast fluorescence PCR instrument (ABI), ultra-trace nucleic acid Protein concentration a...
the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More PUM
Login to View More Abstract
The invention discloses a bluetongue virus-8 detection reagent, detection method and application. The reagent comprises a specific primer pair and a probe, sequences of forward and reverse primers of the primer pair are shown as Seq ID NO.1 and 2, and a sequence of the probe is shown as Seq ID NO.3. In the sequence shown as Seq ID NO.3, fluorescence quenching group-dT is modified by a 32th basic group, a 33th basic group is replaced by a basic group analogue, a fluorescence group-dT is modified by a 35th basic group, and C3Spacer is modified by a 3' terminal. By one-step reverse transcription recombinase polymerase amplification, sensitive, specific and efficient bluetongue virus-8 detection can be realized. By adoption of the reagent, convenient onsite bluetongue virus-8 detection can be realized, and quickness in acquisition of detection results is achieved. An important significance to quick prevention and control of bluetongue virus-8, prevention of epidemic spread and maximum guarantee of production safety is achieved.
Description
technical field [0001] The present application relates to the field of detection of bluetongue virus type 8, in particular to a reagent, detection method and application for detection of bluetongue virus type 8. Background technique [0002] Bluetongue (Bluetongue, BT) is an infectious, non-contact, viral disease transmitted by insect vectors such as Culicoides. The pathogen of the disease is bluetongue virus (Bluetongue virus, BTV), which belongs to the reo Viridae Orbivirus genus. BT primarily affects wild and domesticated ruminants such as sheep, goats, cattle, buffalo, deer, and most African antelope. The average case fatality rate of infected animals is 30%, among which the case fatality rate of sheep can be as high as 80%. BT seriously affects the development of animal husbandry and international trade in many countries. The World Health Organization lists it as a notifiable animal disease, and China lists it as a first-class animal disease. There are many serotypes...
Claims
the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More Application Information
Patent Timeline
20 Sep 2019
Publication
CN110257560A
- IPC
- C12Q1/70; C12Q1/6844; C12N15/11
- CPC
- C12Q1/701; C12Q1/6844; C12Q2521/107; C12Q2531/119; C12Q2521/507; C12Q2563/107; Y02A50/30
- Inventors
- 林彦星; 花群俊