Visible gene chips and kit for porcine pseudorabies virus and/or porcine circovirus type 2

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, fast detection effect

Active Publication Date: 2015-09-09
SICHUAN AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[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.

Method used

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  • Visible gene chips and kit for porcine pseudorabies virus and/or porcine circovirus type 2
  • Visible gene chips and kit for porcine pseudorabies virus and/or porcine circovirus type 2
  • Visible gene chips and kit for porcine pseudorabies virus and/or porcine circovirus type 2

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] Embodiment 1 Detection method of the present invention

[0028] 1. Design of probe genes and construction of plasmid bacteria containing probes

[0029] 1 material

[0030] 1.1 kind of poison

[0031] PRV Fa strain, donated by Chengdu Institute of Biological Products, Ministry of Health;

[0032] PCV2 was isolated from Ziyang disease materials in Sichuan by Zhao Chunrong in our laboratory;

[0033] Inactivated porcine parvovirus vaccine was purchased from Beijing Zhonghai Science and Technology Animal Health Technology Company.

[0034] 1.2 Probe gene recombination plasmid bacteria

[0035] T / GAPDH was synthesized by Yingjun Biological Company.

[0036] 1.3 Main experimental instruments

[0037] Electronic balance: Sartorius BP 310S;

[0038] Ultrapure water meter: Milli Qplus, made in France;

[0039] Ultra-clean bench: SW-CJ-2FD, Suzhou Antai Air Technology Co., Ltd.;

[0040] Constant temperature shaker: vacuum machine: SinBo, Hong Kong;

[0041] 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 PCV2 were 22.3% (2.9% for the vaccine strain and 19.4% for the wild strain) and 78.6% respectively, and 22.3% for PRV+PCV...

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Abstract

The invention discloses gene chips for detecting porcine pseudorabies virus and / or porcine circovirus type 2. The gene chips comprise solid-phase carriers and probes fixed on the solid-phase carriers, wherein the probes comprises gene segments shown as SEQ ID NO:1 and / or 3, and used for detecting the porcine pseudorabies virus and / or the porcine circovirus type 2. The invention further discloses a kit for detecting the porcine pseudorabies virus and / or the porcine circovirus type 2. The gene chips and the kits can be used for effectively detecting the porcine pseudorabies virus and / or the porcine circovirus type 2, and distinguishing the infection of porcine pseudorabies virus vaccine strains from that of porcine pseudorabies virus wild strains, and has the advantages of being high in specificity and sensitivity, and long in storage life; moreover, the time consumption is low, the detection is fast, the detection results can be visibly observed, no expensive equipment is needed, the infection situation of livestock and poultry plague can be rapidly mastered, and the clinical application prospect is good.

Description

technical field [0001] The invention relates to a visual gene chip and a detection method for detecting porcine pseudorabies virus and / or 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) and porcine circovirus type 2 (PCV2) 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 ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/70C12Q1/68C12R1/93
CPCC12Q1/701
Inventor 文心田曹三杰常晓霞黄小波文翼平伍锐张仙马锐杨国淋
Owner SICHUAN AGRI UNIV
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