Visible gene chip and kit for pig pseudorabies virus, porcine parvovirus and porcine circovirus type-II
A technology for porcine pseudorabies virus and porcine circovirus, which is applied in the directions of microorganism-based methods, biochemical equipment and methods, and microorganism determination/inspection, etc. High, observable and intuitive effect
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Embodiment 1
[0028] Embodiment 1 Detection method of the present invention
[0029] 1. Design of probe genes and construction of plasmid bacteria containing probes
[0030] 1 material
[0031] 1.1 kind of poison
[0032] PRV Fa strain, donated by Chengdu Institute of Biological Products, Ministry of Health;
[0033] PCV2 was isolated from Ziyang disease materials in Sichuan by Zhao Chunrong in our laboratory;
[0034] Inactivated porcine parvovirus vaccine was purchased from Beijing Zhonghai Science and Technology Animal Health Technology Company.
[0035] 1.2 Probe gene recombination plasmid bacteria
[0036] T / GAPDH was synthesized by Yingjun Biological Company.
[0037] 1.3 Main experimental instruments
[0038] Electronic balance: Sartorius BP 310S;
[0039] Ultrapure water meter: Milli Qplus, made in France;
[0040] Ultra-clean bench: SW-CJ-2FD, Suzhou Antai Air Technology Co., Ltd.;
[0041] Constant temperature shaker: vacuum machine: SinBo, Hong Kong;
[0042] Constant t...
Embodiment 2
[0170] Example 2 Using the visualized gene chip and kit of the present invention to detect actual samples
[0171] 1. Clinical samples: Spleen, lung, liver, kidney, lymph nodes and other tissues collected from pigs suspected of porcine reproductive disorders in Chengdu, Guang'an, Leshan, Deyang, Anyue, Panzhihua and other places in Sichuan.
[0172] 2. Detection method
[0173] The lesions of the spleen, lung, liver, kidney, and lymph nodes were collected from 103 clinical samples in Sichuan, Chongqing, Fujian, Guizhou, Gansu, etc., and the virus genome was extracted using the Tiangen Blood / Cell / Tissue Genomic DNA Extraction Kit, according to the examples 1 method for detection.
[0174] 3. Test results
[0175] The results are shown in the table below:
[0176] Table 1: Test results of clinical samples
[0177]
[0178]The positive rates of PRV, PPV, and PCV2 were 22.3% (2.9% for the vaccine strain, 19.4% for the wild strain), 8.7%, and 78.6%, respectively; PRV+PPV was...
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