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Visible gene chip and kit for pig pseudorabies virus, porcine parvovirus and porcine circovirus type-II
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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
Active Publication Date: 2015-08-26
SICHUAN AGRI UNIV
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[0003] At present, the diagnosis of these three diseases mostly adopts isolation and identification and conventional serological methods, which takes a long time. There is also a method for identification by PCR technology, but there is no method that is highly operable and can simultaneously identify the three viruses.
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Embodiment 1
[0028] Embodiment 1 Detection method of the present invention
[0029] 1. Design of probe genes and construction of plasmidbacteria 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.
[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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Abstract
The invention discloses a visible genechip for a pig pseudorabiesvirus, porcine parvovirus and porcine circovirus type-II. The visible genechip comprises a solid phase carrier and a probe fixed on the solid phase carrier, wherein the probe comprises gene segments as shown in SEQ ID NO:1, 3, and 4; and the gene segments are used for respectively detecting the pig pseudorabiesvirus, the porcine parvovirus and the porcine circovirus type-II. The invention further discloses a kit for detecting the pig pseudorabiesvirus, the porcine parvovirus and the porcine circovirus type-II. By adopting the gene chip and the kit disclosed by the invention, pathogens of the pig pseudorabies virus, the porcine parvovirus and the porcine circovirus type-II can be effectively detected, infection caused by vaccine strains and wild strains of the pig pseudorabies virus can be distinguished, the gene chip and the kit have the characteristics of good specificity, high sensitivity and long preservation period, are short in time, rapid in detection, visible in detection result observation and good in clinical application prospect, no expensive equipment is needed, and the infection situation of livestock diseases can be rapidly handled.
Description
technical field [0001] The invention relates to a visual gene chip and a detection method for detecting porcine pseudorabies virus, porcine parvovirus and porcine circovirus type 2. Background technique [0002] With the development of my country's pig industry in the direction of intensification and scale, large-scale pig farms have frequent epidemics and mixed infections of various pathogens, which have caused certain losses to the pig industry. Among them, porcine pseudorabies virus (PRV), porcine parvovirus (PPV) and porcine circovirus type 2 (PCV2) are three important disease pathogens that cause reproductive disorders in pigs. The clinical symptoms of these three diseases are similar, and differential diagnosis is required. [0003] At present, the diagnosis of these three diseases mostly adopts isolation and identification and conventional serological methods, which take a long time. There is also a method for identification by PCR technology, but there is no method ...
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