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Visual gene chip and kit used for detecting porcine epidemic encephalitis b viruses and/or porcine reproductive and respiratory syndrome viruses

A technology of Japanese encephalitis virus and respiratory syndrome, applied in the direction of combinatorial chemistry, chemical library, organic compound library, etc., can solve the problems of high price, high cost, unfavorable promotion and use, etc., and achieve low cost, short time consumption, Promising results for clinical application

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 publication number CN104293976A discloses a chip for detecting porcine encephalitis Japanese encephalitis virus and porcine reproductive and respiratory syndrome virus. Requires an expensive laser co-polymerization scanner to scan and analyze the results, the cost is too high to be conducive to popularization

Method used

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  • Visual gene chip and kit used for detecting porcine epidemic encephalitis b viruses and/or porcine reproductive and respiratory syndrome viruses
  • Visual gene chip and kit used for detecting porcine epidemic encephalitis b viruses and/or porcine reproductive and respiratory syndrome viruses
  • Visual gene chip and kit used for detecting porcine epidemic encephalitis b viruses and/or porcine reproductive and respiratory syndrome viruses

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Experimental program
Comparison scheme
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 PRRSV-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] PRRSV: porcine reproductive and respiratory syndrome live vaccine (JXA1-R strain), purchased from China Animal Husbandry Co., Ltd.; Japanese encephalitis virus: JEV SCYA20120201 strain, obtained from a case with abortion symptoms in a pig farm in Ya'an. 1.1.2 Negative samples

[0176] Negative samples were taken from lymph nodes, lungs and other tissues of healthy pigs from a pig farm in Ya'an.

[0177] 1.2 Clinical samples

[0178] 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.

[0179] 1.3 Main reagents

[0180] Total RNA extrac...

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Abstract

The invention discloses a visual gene chip and kit used for detecting porcine epidemic encephalitis b viruses and / or porcine reproductive and respiratory syndrome 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 visual gene chip and a kit for detecting porcine epidemic Japanese encephalitis virus and / or porcine reproductive and respiratory syndrome virus. Background technique [0002] Mixed infection with multiple pathogens is a common phenomenon in current pig farm diseases. Porcine reproductive and respiratory syndrome and porcine epidemic encephalitis B have similar disease symptoms and are often mixed infections. Therefore, early differential diagnosis and timely understanding of the epidemic situation are particularly important for controlling the epidemic 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 es...

Claims

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Application Information

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IPC IPC(8): C12Q1/70C12Q1/68C40B40/06C40B50/18C12R1/93
CPCC12Q1/701
Inventor 文心田黄小波张仙曹三杰文翼平伍锐常晓霞马锐杨国淋
Owner SICHUAN AGRI UNIV
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