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High-flux fast detecting method for virus pathogen

A detection method and high-throughput technology, which is applied in the field of high-throughput rapid screening and detection of porcine epidemic virus, can solve the problems of low sensitivity and specificity, inaccurate detection results, laborious and other problems, and achieve high sensitivity and specificity , easy operation and stable test results

Inactive Publication Date: 2008-07-02
SHANXI LIFEGEN
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Problems solved by technology

[0005] The purpose of the present invention is to provide a high-throughput rapid screening and detection method for pathogens, which solves the technical problems of time-consuming, laborious, low sensitivity and specificity, and inaccurate detection results in the swine epidemic virus detection method in the background technology

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  • High-flux fast detecting method for virus pathogen
  • High-flux fast detecting method for virus pathogen
  • High-flux fast detecting method for virus pathogen

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Embodiment Construction

[0039] The virus pathogen high-throughput rapid screening and detection method adopted in the present invention has the following steps:

[0040] 1. Prepare chip:

[0041] The probe design and chip spotting in the chip preparation of the present invention can adopt known techniques.

[0042] (1.1) Design of probes: Design a highly specific 60bp oligonucleotide probe according to the conserved sequence of the viral genome.

[0043] (1.2) Chip spotting: first carry out the surface modification treatment of the substrate, then fix the probes on the substrate according to the designed array, prepare an oligonucleotide chip, and perform post-processing on the chip to obtain the oligonucleotide chip.

[0044] 2. Processing of samples to be tested:

[0045] The sample processing of the invention adopts the method of indirect labeling, the labeling efficiency is high, the detection sensitivity is improved, and the cost is low.

[0046] (2.1) Design and synthesis of primers: obtain...

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Abstract

A high-throughput method for rapid detection of virus pathogens comprises the following steps of: (1) designing probes and spotting on a chip to obtain an oligonucleotide chip; (2) designing and synthesizing primers to obtain primers corresponding to a detected pathogen; (3) performing synchronous asymmetric PCR amplification to a detected sample with the primers corresponding to the detected pathogen to obtain a single-stranded DNA product specifically complementary with the oligonucleotide; (4) labeling a fluorescent substance on the single-stranded DNA product to obtain a fluorescent-labeled single-stranded DNA product; (5) hybridizing with the oligonucleotide chip, and cleaning the chip; and (6) scanning with a fluorescent scanner to obtain virus detection and analysis results. The invention solves the time-consuming, labor-consuming, low sensibility and specificity and inaccurate detection result problems in the detection method of virus causing porcine epidemic diseases in the prior art. The invention can be used for rapid, accurate and efficient detection of multiple genes.

Description

technical field [0001] The invention relates to a screening and detection method for virus pathogens, in particular to a high-throughput rapid screening and detection method for porcine epidemic disease virus. Background technique [0002] Oligonucleotide Microarrays (Oligonucleotide Microarrays) are designed and synthesized oligonucleotide probes of a dozen to dozens of bases that are regularly arranged and fixed on the support through a spotting instrument, and then combined with fluorescent light. The labeled target sequence is hybridized under certain conditions according to the principle of base pairing. After washing, the chip is scanned by the laser confocal fluorescence detection system, and the data is compared and analyzed by the computer. One test can quickly and accurately detect multiple genes. , Efficient detection. [0003] Porcine parvovirus (PPV), porcine circovirus (PCV), and swine fever virus (CSFV) are common pathogens that cause disease in pigs, and oft...

Claims

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Application Information

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IPC IPC(8): C12Q1/70C12Q1/68
Inventor 李铮陈超高金拽王小强祁会彩黄鹤
Owner SHANXI LIFEGEN
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