Primer group used for detecting canine parvovirus and application of primer group used for detecting canine parvovirus
A technology of canine parvovirus and primer set, applied in the biological field, can solve the problems of detection of canine parvovirus, such as specificity and poor sensitivity, and achieve the effects of high sensitivity, strong practicability and strong specificity
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
Method used
Image
Examples
Embodiment 1
[0031] Embodiment 1, be used for the design of the primer set of detecting canine parvovirus
[0032] For the VP2 gene sequence in the canine parvovirus genome, download the canine parvovirus gene sequence registered in GenBank, use DNASTAR's MegAlign software for analysis and comparison, and select VP 2 For the conserved region fragment in the front of the gene (GenBankMN561321.1), use the online website for LAMP primer design (http: / / primerexplorer.jp / e / ) to design primers and design a specific LAMP primer set, which includes outer primers and inner primers pairs and circular primer pairs. Among them, the outer primer pair is composed of F3 and B3, the inner primer pair is composed of FIP and BIP, and the loop primer pair is composed of loopB and loopF. As shown in Table 1.
[0033] Table 1
[0034]
Embodiment 2
[0035] Embodiment 2, detection canine parvovirus specificity
[0036] 1. Strains: canine parvovirus and canine adenovirus were isolated from the Beijing Institute of Animal Husbandry and Veterinary Medicine, Chinese Academy of Agricultural Sciences. The clinically collected canine parvovirus suspected disease materials were inoculated into cat kidney cells (CRFK cells), and canine parvovirus was isolated after 3 passages. Virus. Canine infectious hepatitis disease materials were inoculated into canine kidney cells (MDCK cells), and canine adenovirus was isolated after three passages. Canine coronavirus is Weijia 8 vaccine strain for dogs, which was purchased from Zoetis, USA. The encephalomyocarditis virus was preserved by the Veterinary College of China Agricultural University, and the canine distemper virus was the vaccine strain CDV-11, which was purchased from Qilu Animal Health Products Co., Ltd.
[0037] 2. Extraction of viral nucleic acid: use AxyPrep™ Body Fluid Vira...
Embodiment 3
[0047] Embodiment 3, detection canine parvovirus sensitivity
[0048] The positive plasmid standard prepared in Example 2 was serially diluted 10 times, and the copy number was 3.4×10 1 Copy number / μl to 3.4×10 5 The copy number / μl samples of five gradients were subjected to LAMP reaction to determine the detection sensitivity, and the experimental operation was repeated once.
[0049] Sensitivity results of the LAMP method, the prepared positive standard plasmid standard was serially diluted 10 times, and the detection sensitivity of the canine parvovirus ring-mediated isothermal amplification method was determined to be 3.4×10 1 Copy number / μl, its sensitivity is 100 times that of ordinary PCR. Such as Figure 4 As shown, it is the fluorescent signal diagram of the sensitivity pattern of the embodiment of the present invention, such as Figure 5 Shown is the melting curve of the canine parvovirus LAMP susceptibility experiment of the embodiment of the present invention. ...
PUM
Login to View More Abstract
Description
Claims
Application Information
Login to View More