Group B streptococcus fluorescent quantitative PCR detection kit
A fluorescent quantitative and streptococcal technology, applied in the determination/testing of microorganisms, DNA/RNA fragments, biochemical equipment and methods, etc., can solve the problems of urine, feces, human genomic DNA, clotrimazole suppositories, miconazole Suppositories, nystatin ointment, Imodium or compound carrageenate suppositories cannot meet the problems of fast, early diagnosis, accuracy and sensitivity, etc., and achieve the effect of reducing synthesis cost, increasing success rate and strong sensitivity
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
Method used
Image
Examples
Embodiment 1
[0041] 1. Design 4 sets of GBS amplification primer pairs, GBS detection probes, internal reference primer pairs and internal reference probes, the nucleotide sequences are shown in Table 1, and study the detection of complex samples with different primers and probes.
[0042] Table 1 Nucleotide sequences of primers and probes
[0043]
[0044] 2. Prepare 4 kinds of kits, each kit contains only one of the 4 sets of primers and probes A, B, C, and D. Except for the primers and probes, the rest of the components are the same, and kits A, B, and D are obtained respectively. B, C, D.
[0045] The preparation steps of the kit are as follows:
[0046] A fluorescent quantitative PCR detection kit for group B streptococci comprises a set of fluorescent quantitative PCR detection primers and probes for group B streptococcus, DNA extraction solution, PCR reaction solution, positive control solution and negative control solution. Wherein said Group B Streptococcus fluorescent quanti...
Embodiment 2
[0071] Use kit A to detect group B streptococcus in clinical samples, the steps are as follows:
[0072] 1. Extract sample DNA
[0073] 1. Preparation before detection
[0074] Take out the reagents from -20°C 15-30 minutes in advance, thaw the reagents, vortex each component for 20-40s, and briefly centrifuge for 2-6s; prepare a water bath and set the temperature to 100°C.
[0075] 2. Preparation of DNA Extraction Solution
[0076] Recipe for 1mL DNA extraction solution: 1×Tris-EDTA Buffer 990μL+Triton X-100 10μL.
[0077] Mix the above-mentioned 1×Tris-EDTA Buffer and Triton X-100, heat at 30-40°C, and stir magnetically for 15-30 minutes until the glue block is completely dissolved. Among them, Triton X-100 is a very viscous liquid, which should be stirred until it is fully dissolved and mixed evenly, and the preparation should be completed within 1 hour. The obtained DNA extract is a colorless and clear liquid with a pH value of 7.5-8.2 and stored in a refrigerator (2-8...
Embodiment 3
[0109] Specific detection of kit A:
[0110] Select clinical common pathogens Group B Streptococcus, Streptococcus pneumoniae, Streptococcus pyogenes, Streptococcus thermophilus, Streptococcus mutans, Streptococcus pyogenes, Lactobacillus acidophilus, Lactobacillus reuteri, Staphylococcus epidermidis, Escherichia coli Escherichia DH5α, Candida albicans, respectively extract DNA according to the method in Example 2, prepare PCR reaction solution, and detect each bacterial strain respectively according to the detection method of Group B Streptococcus in Example 2, and use Group B Streptococcus Standard strains were used as positive controls.
[0111] The result is as figure 2 As shown, only the standard and clinical strains of group B Streptococcus were positive, indicating that the kit A has high specificity, and the kit A will not cross-react with various common clinical pathogens.
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