Kit and detection method for detecting multiple common duck-origin viruses
A kit and virus technology, applied in biochemical equipment and methods, microbe determination/inspection, DNA/RNA fragments, etc., can solve the problems of time-consuming and labor-intensive isolation and culture, low sensitivity of immunoassay, etc., and achieve good room temperature preservation Stability, lower fatality rate, and shorter detection time
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
Method used
Image
Examples
Embodiment 1
[0036] Example 1 A kit for detecting a variety of common duck-derived viruses
[0037] In this embodiment, multiplex fluorescent PCR method is used to detect a variety of common duck-derived virus kits, including reaction reagents, positive control substances and negative control substances.
[0038] (1) Reagent: take a 10mL volumetric flask, add 0.5mol / L Trizma® HCl 96μL, 0.5mol / L Trizma® Base 504μL, 1 mol / L MgCl respectively 235 μL, 1mol / L KCl 1200 μL, DMSO 100 μL, dNTPs 135 μL, 50 μmol / L duck hepatitis A type 1 upstream and downstream primers 120 μL each, 50 μmol / L duck hepatitis A type 1 probe 30 μL, 50 μmol / L duck hepatitis A Hepatitis A type 3 upstream and downstream primers 120 µL each, 50 µmol / L Duck hepatitis A type 3 probe 30 µL, 50 µmol / L duck astrovirus upstream and downstream primers 120 µL each, 50 µmol / L duck astrovirus probe 30 µL, 50 µmol / L 120 μL of upstream and downstream primers for duck paramyxovirus, 30 μL of 50 μmol / L duck paramyxovirus probe, 1 μL of H...
Embodiment 2
[0052] Example 2 A detection method for detecting multiple common duck-derived viruses
[0053] The multiple fluorescent PCR detection method of multiple common duck-derived viruses in the present embodiment comprises the following specific steps:
[0054] 1. Main kit: the kit of Example 1 is used.
[0055] 2. Nucleic acid extraction: Use a qualified nucleic acid extraction kit to extract nucleic acid from the sample to be tested. After extraction, use a trace ultraviolet spectrophotometer to measure the nucleic acid purity. The OD260 / OD280 should be between 1.6-2.0.
[0056] 3. Load the sample on the machine
[0057] (1) Dissolution of the positive control substance: Take 250 μL of the negative control substance and add it to the positive control lyophilized powder, mix well and set aside.
[0058] (2) Adding samples: Take out the eight-tube strip of reaction reagents, add 25 μL sample nucleic acid, 25 μL positive control substance or 25 μL negative control substance respec...
Embodiment 3
[0066] Example 3 Specificity, repeatability and sensitivity detection
[0067] 1. Specific detection
[0068] (1) Specific samples: Prepare 10 specific samples, among which specific samples 1-4 are normal saline, sample 5 is a chicken Newcastle disease virus sample, sample 6 is a classic reovirus sample, and sample 7 is a new type of reovirus Virus samples, sample 8 is a duck Tembusu virus sample, sample 9 is a duck circovirus sample, and sample 10 is a duck adenovirus sample.
[0069] (2) Experimental process: Using the kit of Implementation 1, the multiplex fluorescent PCR detection method in Example 2 was used to detect the above 10 specific samples respectively. The detection results are shown in Table 1 and figure 1 Shown: figure 1 In , the ordinate represents the fluorescence value, and the abscissa represents the CT value.
[0070] Table 1 Detection results of four fluorescence channels of specific samples 1-10
[0071]
[0072] (3) Experimental results: from Tab...
PUM
Login to View More Abstract
Description
Claims
Application Information
Login to View More