Method for detecting bovine viral diarrhea viruses as well as preparation method and use method of reverse transcription loop-mediated isothermal amplification (RT-LAMP) reaction kit
A bovine viral diarrhea, ring-mediated isothermal technology, which is applied in the directions of microorganism-based methods, biochemical equipment and methods, and microorganism determination/inspection, etc., can solve problems such as discomfort and high cost, and achieve cost reduction and convenient detection. Fast, highly sensitive effects
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
Method used
Image
Examples
specific Embodiment approach 1
[0026] Embodiment 1: The preparation method of the reverse transcription loop-mediated isothermal amplification reaction kit for detecting bovine viral diarrhea virus in this embodiment is realized in the following steps: 1. Retrieve the gene sequences of all bovine viral diarrhea viruses from the gene database , through Clustalw software analysis to find out the specific conserved target DNA sequence, and then use the Primer Explorer 4 software to design 6 primers; 2. Prepare the reverse transcription loop-mediated isothermal amplification reaction solution according to the obtained 6 primers, and then complete the detection of cattle 病毒性腹泻病毒的逆转录环介导等温扩增反应试剂盒的制备;其中步骤一中保守靶DNA序列为:TGAGGGACAAATCCTCCTTAGCGAAGGCCGAAAAGAGGCTAGCCATGCCCTTAGTAGGACTAGCAAAACAAGGAGGGTAGCAACAGTGGTGAGTTCGTTGGATGGCTGAAGCCCTGAGTACAGGGTAGTCGTCAGTGGTTCGACGCTTTGTGCGACAAGCCTCGAGATGCCACGTGGACGAGGGCATGCCCACAGCACATCTTAACCTGAGCGGGGGTCGTTCAGGTGAAAACGGTTTAACCAACCGCTACGAATACAGCCTGATAGGGTGCTGCAGAGGCCCACTGTATCGCTACTAAAAATCTCT...
specific Embodiment approach 2
[0037] Specific embodiment two: the use method of the reverse transcription loop-mediated isothermal amplification reaction kit for detection of bovine viral diarrhea virus in this embodiment: one, get 5 μ l of the RNA extract containing bovine viral diarrhea virus sample, add 20 μ l reverse transcription The loop-mediated isothermal amplification reaction solution was evenly mixed and placed at 62°C for reverse transcription loop-mediated isothermal amplification reaction for 1 hour; 2. After the amplification reaction, centrifuged at 10000r / min for 1min, there was precipitation at the bottom of the reaction tube , then there is bovine viral diarrhea virus in the sample to be tested, and there is no precipitation at the bottom of the reaction tube, then there is no bovine viral diarrhea virus in the sample to be tested.
[0038] In this embodiment, a negative control group can be set up for detection, and the negative control group uses water instead of RNA extract, and the re...
specific Embodiment approach 3
[0039] Specific embodiment three: the use method of the reverse transcription loop-mediated isothermal amplification reaction kit for detection of bovine viral diarrhea virus in this embodiment: 1. Take 5 μl of the RNA extract containing bovine viral diarrhea virus sample, add 20 μl reverse transcription The loop-mediated isothermal amplification reaction solution was uniformly mixed and placed at 62°C for reverse transcription loop-mediated isothermal amplification reaction for 1 hour; 2. After the amplification reaction, 5 μl of 10000×SYBRGREEN I was added, and then placed under a UV lamp. If the reaction liquid turns green, there is bovine viral diarrhea virus in the sample to be tested, and if the reaction liquid is orange, then there is no bovine viral diarrhea virus in the sample to be tested.
[0040] In this embodiment, a negative control group can be set up for detection, and the negative control group uses water instead of RNA extract, and the rest of the components a...
PUM
Abstract
Description
Claims
Application Information
- R&D Engineer
- R&D Manager
- IP Professional
- Industry Leading Data Capabilities
- Powerful AI technology
- Patent DNA Extraction
Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.
© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com