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Visualized gene chip and kit for porcine pseudorabies virus and / or porcine parvovirus
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A technology of porcine pseudorabies virus and pseudorabies virus, which is applied in the field of visualized gene chips, can solve the problems of long time required and achieve the effect of short time consumption, high sensitivity and intuitive observation
Inactive Publication Date: 2015-10-14
SICHUAN AGRI UNIV
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[0003] At present, the diagnosis of these two 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 two viruses.
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Embodiment 1
[0026] Embodiment 1 Detection method of the present invention
[0027] 1. Design of probe genes and construction of plasmidbacteria containing probes
[0028] 1 material
[0029] 1.1 kind of poison
[0030] PRV Fa strain, donated by Chengdu Institute of Biological Products, Ministry of Health;
[0031] PCV2 was isolated from Ziyang disease materials in Sichuan by Zhao Chunrong in our laboratory;
[0032] Inactivated porcine parvovirus vaccine was purchased from Beijing Zhonghai Science and Technology Animal Health Technology Company.
[0038] Ultra-clean bench: SW-CJ-2FD, Suzhou Antai Air Technology Co., Ltd.;
[0039] Constant temperature shaker: vacuum machine: SinBo, Hong Kong;
[0040] Constant t...
Embodiment 2
[0160] Example 2 Using the visualized gene chip and kit of the present invention to detect actual samples
[0161] 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.
[0162] 2. Detection method
[0163] 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.
[0164] 3. Test results
[0165] The results are shown in the table below:
[0166] Table 1: Test results of clinical samples
[0167]
[0168] The positive rates of PRV and PPV were 22.3% (2.9% for the vaccine strain and 19.4% for the field strain) and 8.7% respectively, and 8.7% for PRV+PPV. ...
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Abstract
The invention discloses a genechip for detecting porcine pseudorabiesvirus and / or porcine parvovirus. The genechip comprises a solid phase carrier and a probe immobilized on the solid phase carrier; and the probe comprises gene fragments shown as SEQ ID NO: 1 and / or 3 for detecting pseudorabiesvirus and / or porcine parvovirus respectively. The invention discloses a kit for detecting porcine pseudorabiesvirus and / or porcine parvovirus. The gene chip and kit provided by the invention can effectively detect porcine pseudorabies virus and porcine parvovirus, also distinguish the vaccine strain infection from wild strain infection of the porcine pseudorabies virus, have the characteristics of good specificity, high sensitivity and long shelf life, short time consumption, rapid detection, visual observation of the detection results and no requirement of expensive equipment. The gene chip and kit can quickly grasp the infection of animal diseases, and have good prospects for clinical application.
Description
technical field [0001] The invention relates to a visual gene chip and a detection method for detecting porcine pseudorabies virus and / or porcine parvovirus. 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) and porcine parvovirus (PPV) are two important disease pathogens that cause reproductive disorders in pigs. The clinical symptoms of these two diseases are similar, and a differential diagnosis is required. [0003] At present, the diagnosis of these two 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 that is highly operable and can simultaneously identify the tw...
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