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Visual gene chip and kit used for detecting porcine epidemic encephalitis b viruses and/or hog cholera viruses

A technology of Japanese encephalitis virus and swine fever virus, applied in the direction of combinatorial chemistry, chemical library, organic compound library, etc., can solve the problems of unfavorable promotion and use, high price, high cost, etc., and achieve good clinical application prospects, low cost, short time effect

Inactive Publication Date: 2015-11-04
SICHUAN AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] The patent application document with the publication number: CN104293981A discloses a chip for detecting porcine encephalitis Japanese encephalitis virus and / or swine fever virus, which uses fluorescently labeled primers for detection. Expensive laser converging scanner to scan and analyze the results, the cost is too high, not conducive to popularization

Method used

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  • Visual gene chip and kit used for detecting porcine epidemic encephalitis b viruses and/or hog cholera viruses
  • Visual gene chip and kit used for detecting porcine epidemic encephalitis b viruses and/or hog cholera viruses
  • Visual gene chip and kit used for detecting porcine epidemic encephalitis b viruses and/or hog cholera viruses

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Experimental program
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Effect test

Embodiment 1

[0041] Embodiment 1 The preparation of the visualized gene chip of the present invention and its detection method

[0042] 1. Materials and instruments

[0043] The same experimental materials and instruments as mentioned above.

[0044] 2 Preparation and performance testing of the gene chip of the present invention

[0045] 2.1 Preparation of primers and probes

[0046] In this study, the virus-specific DNA fragment immobilized on the amino substrate is called the probe, and the sample to be tested labeled with biotin is called the target gene.

[0047] 2.1.1 Synthesis of primers

[0048] The primers were synthesized by Shanghai Sangon Bioengineering Co., Ltd., and two reverse primers were synthesized, one unlabeled and one 5' end labeled with Biotin.

[0049] Table 1 Target gene amplification primers

[0050]

[0051] Table 2 Mapping gene primers

[0052]

[0053] 2.1.2 Probe genes and positioning genes

[0054] JEV-C (SEQ ID NO: 1):

[0055] Gene size and sequen...

experiment example 1

[0171] Experimental example 1 CSFV-JEV visual co-examination gene chip detection of clinical samples

[0172] 1 material

[0173] 1.1 Positive and negative samples

[0174] 1.1.1 Positive samples

[0175] CSFV: live swine fever vaccine (cell source), purchased from Shandong Yakang Pharmaceutical; Japanese encephalitis virus: JEV SCYA20120201 strain, isolated from a case with abortion symptoms in a pig farm in Ya'an.

[0176] 1.1.2 Negative samples

[0177] Negative samples were taken from lymph nodes, lungs and other tissues of healthy pigs from a pig farm in Ya'an.

[0178] 1.2 Clinical samples

[0179] From January 2014 to January 2015, 102 clinical samples from Meishan, Mianyang, Leshan, Shuangliu, Dazhou, Xichang, Chongqing and other places in Sichuan were collected from the tissues of sick pigs with clinical manifestations of suspected porcine reproductive disorder or a miscarriage.

[0180] 1.3 Main reagents

[0181] Total RNA extraction kit (DP419) was purchased ...

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Abstract

The invention discloses a visual gene chip and kit used for detecting porcine epidemic encephalitis b viruses and / or hog cholera viruses. With the visual gene chip prepared by using a specific method, detection results can be visually seen with naked eyes, and an expensive laser confocal scanner is not needed; and the visual gene chip and kit have the advantages of low cost, high sensitivity which is almost equal to sensitivity of conventional fluorescence labeling methods, good specificity, short detection time, rapid detection and good application prospects.

Description

technical field [0001] The invention relates to a visualized gene chip and a kit for detecting porcine epidemic Japanese encephalitis virus and / or swine fever virus. Background technique [0002] Mixed infection with multiple pathogens is a common phenomenon in current pig farm diseases. The disease symptoms caused by these two pig viruses are similar and often mixed infection. They are two important viruses that endanger the pig industry at present. Therefore, early differential diagnosis and timely understanding of the epidemic situation are particularly important for controlling the prevalence of these two diseases. At present, the isolation of pathogens and conventional serological methods are mostly used in the diagnosis of these two diseases, but it takes a long time. Recently, the isolation and identification of pathogens, immune colloidal gold technology, enzyme-linked immunosorbent assay, serum neutralization test, polymerase chain reaction, etc. have been establis...

Claims

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Application Information

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IPC IPC(8): C12Q1/70C12Q1/68C40B40/06C40B50/18C12R1/93
CPCC12Q1/701
Inventor 文心田黄小波欧阳达曹三杰伍锐文翼平常晓霞杨国淋马锐
Owner SICHUAN AGRI UNIV
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