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Oligonucleotide chip for synchronous detection of four porcine diarrhea viruses, and application of oligonucleotide chip

An oligonucleotide, simultaneous detection technology, applied in the field of molecular biology, can solve the problem of high-sensitivity oligonucleotide chips that have not been detected simultaneously

Active Publication Date: 2018-09-14
SICHUAN AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, there is no report on the preparation of oligonucleotide chips for porcine diarrhea, and there is no high-sensitivity oligonucleotide chip for simultaneous detection of four porcine diarrhea viruses.

Method used

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  • Oligonucleotide chip for synchronous detection of four porcine diarrhea viruses, and application of oligonucleotide chip
  • Oligonucleotide chip for synchronous detection of four porcine diarrhea viruses, and application of oligonucleotide chip
  • Oligonucleotide chip for synchronous detection of four porcine diarrhea viruses, and application of oligonucleotide chip

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Embodiment 1

[0042] Example 1 Construction of PEDV-TGEV-GAR-PDCoV oligonucleotide co-detection chip of the present invention

[0043] 1. Biomaterials

[0044] 1.1 Plasmid

[0045] Recombinant plasmids pMD-PS, pMD-PM, pMD-TS, pMD-TN, pMD-VP7, pMD-NSP4, pMD-N, pMD-M (for the preparation method, see "2 Construction of recombinant plasmids for target genes"), pMD- λ (purchased from Bao Bio (Dalian) Biotechnology Co., Ltd.), and the plasmids were constructed and preserved by our laboratory.

[0046] 1.2 Main reagents:

[0047] Total RNA extraction kit, blood / tissue / cell genome extraction kit: Tiangen Biochemical Technology Company;

[0048] Small amount of plasmid extraction kit: Omega bio-tek company;

[0049] PrimeScript TM RT Reagent Kit, positive quality control (λDNA): Treasure Bioengineering (Dalian) Co., Ltd.;

[0050] Aldehyde-based substrate, gene chip hybridization buffer, optical grade aldehyde-based substrate: Shanghai Bio-engineering Co., Ltd.;

[0051] Optical grade ald...

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Abstract

The invention provides an oligonucleotide chip for synchronous detection of four porcine diarrhea viruses. The oligonucleotide chip comprises a solid phase carrier and oligonucleotide probes fixed onthe solid phase carrier, wherein the oligonucleotide probes comprise a probe shown in SEQ ID No. 1-2, a probe shown in SEQ ID No. 3-4, a probe shown in SEQ ID No. 5-6, and a probe shown in SEQ ID No.7-8; the probes are respectively used for detecting porcine epidemic diarrhea virus (PEDV), porcine transmissible gastroenteritis virus (TGEV), porcine group A rotavirus (GAR) and porcine delta coronavirus (PDCoV). The invention also provides a kit and a method for the synchronous detection of the four porcine diarrhea viruses. The chip and the method can realize the high flux co-detection of thefour porcine diarrhea viruses, i.e., PEDV, TGEV, GAR and PDCoV, and can be applied to mass detection of clinical diseases, thus having a wide application prospect.

Description

technical field [0001] The invention belongs to the field of molecular biology, and in particular relates to an oligonucleotide chip for synchronous detection of four porcine diarrhea viruses and an application thereof. Background technique [0002] In recent years, the occurrence and prevalence of porcine infectious diarrhea disease have become increasingly serious, causing incalculable economic losses to the majority of farmers. Porcine epidemic diarrhea virus (PEDV), porcine transmissible gastroenteritis virus (TGEV), porcine group A rotavirus , GAR). The appearance characteristics of the diseases caused by these three types of viruses are basically similar after the occurrence, and they all manifest as vomiting, watery jet diarrhea and dehydration, especially in sucking piglets with severe disease and high mortality. Porcine deltacoronavirus (Porcine Deltacoronavirus, PDCoV) was first reported in pig farms in Hong Kong in 2012, and then reported in Ohio and Illinois in...

Claims

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Application Information

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IPC IPC(8): C12Q1/70C12Q1/6837C40B40/06C12N15/11
CPCC12Q1/6837C12Q1/701C40B40/06Y02A50/30
Inventor 黄小波文心田曹三杰滑翔文翼平赵勤伍锐
Owner SICHUAN AGRI UNIV
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