Pig virus gene chip and detection method thereof

A virus gene and chip detection technology, applied in the field of microbial detection, can solve the problems of false positive judgment, limited sensitivity, time-consuming, etc., achieve high-throughput accurate detection, facilitate combination, and avoid false positive effects

Active Publication Date: 2012-07-25
BEIJING UNIV OF AGRI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the traditional methods used for the detection of infectious viruses in pigs generally include: virus isolation method, electron microscope observation, polymerase chain reaction, indirect immunofluorescence test, immunohistochemical technique, enzyme-linked immunosorbent assay, etc. Many defects: The advantage of the polymerase chain reaction (PCR) method is high sensitivity, but the detection of PCR results by gel ele

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  • Pig virus gene chip and detection method thereof
  • Pig virus gene chip and detection method thereof
  • Pig virus gene chip and detection method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0010] A porcine virus gene chip, the gene chip includes probes immobilized on a matrix carrier, and the probes are characteristic fragments selected from the following viruses: classical swine fever virus (CSFV), porcine reproductive and respiratory syndrome virus ( PRRSV), pseudorabies virus (PRV), porcine circovirus (PCV), porcine parvovirus (PPV). The matrix carrier is a glass sheet that has undergone aldehyde treatment. The names of the probes in this example are detailed in Table 4, Table 6, and Table 9, and the nucleotide sequences of each probe are detailed in Table 10.

[0011] A method for porcine virus gene chip detection of porcine virus, said gene chip includes probes fixed on a matrix carrier, said probes are selected from the characteristic fragments of the following viruses: classical swine fever virus (CSFV), porcine reproductive and Respiratory Syndrome Virus (PRRSV), Pseudorabies Virus (PRV), Porcine Circovirus (PCV), Porcine Parvovirus (PPV); the method in...

Embodiment 2

[0020] This example is the preparation method of the gene chip in Example 1. The steps of this method are as follows:

[0021] A. Select the appropriate slide. The glass slides should meet the following conditions: choose an optical glass slide as the raw material for the preparation of the aldehyde-based substrate, with a size of 75.5 mm in length, 25.2 mm in width, and 1.0 mm in thickness; The requirements of the instrument for spotting substrates require that the error of length and width is less than or equal to plus or minus 0.2 mm, and the error of thickness is less than or equal to plus or minus 0.05 mm; at the same time, the appearance is required to be free from scratches and defects.

[0022] The reason why glass slides are selected in this embodiment is that the gene chip analysis has high requirements on the surface of the substrate, and the preparation of the surface is a strict process. The quality of surface treatment determines the degree of immobilization of...

Embodiment 3

[0046] This example is a preferred solution based on Example 1, using a gene chip detection system using random primer PCR method. This example uses the gene chip in Example 2. A method for porcine virus gene chip detection of porcine virus, said gene chip includes probes immobilized on a matrix carrier, said probes include characteristic fragments selected from the following viruses: classical swine fever virus (CSFV), porcine reproductive With Respiratory Syndrome Virus (PRRSV), Pseudorabies Virus (PRV), Porcine Circovirus (PCV), Porcine Parvovirus (PPV), the method includes the steps of:

[0047] A. Extracting viral nucleic acid to obtain a sample to be tested. In this example, classical swine fever virus (CSFV), porcine reproductive and respiratory syndrome virus (PRRSV), pseudorabies virus (PRV), porcine circovirus (PCV), porcine parvovirus (PPV), and porcine encephalitis virus (JEV), bovine viral diarrhea virus (BVDV) 7 kinds of virus standard nucleic acid (DNA / RNA) as...

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Abstract

The invention provides a pig virus gene chip and a detection method thereof. The gene chip comprises a probe fixedly arranged on a substrate carrier, wherein the probe is selected from the characteristic segments of the following viruses: a classical swine fever virus (CSFV), a porcine reproductive and respiratory comprehensive virus (PRRSV), a pseudorabies virus (PRV), a porcine circovirus (PCV) and a porcine parvovirus (PPV). The types of the viruses detected by the method are various and the viruses cover common porcine viruses substantially. Random primer polymerase chain reaction (PCR) and multiplex-PCR are adopted to mark, so that false positive which is easy to cause when a PCR result is detected by gel electrophoresis is avoided, and high-flux accurate detection with short time is realized. The substrate carrier is a glass sheet which is subjected to aldehyde treatment, so that the combination between the probe and a target is facilitated, and higher noise is not brought to detection.

Description

Technical field [0001] The invention is the field of microbial testing, which involves a pork virus gene chip and its detection method.The genetic chip includes a probe fixed on the substrate carrier.(PRV), pig ring virus (PCV), pork fine virus (PPV). Background technique [0002] Animal quarantine issues have always been a hot issue that the international community and governments have paid attention to. Especially after joining the WTO in my country, due to the influence of technical trade measures, the hygiene quality requirements for domestic livestock products are getting higher and higher. Many, such as pathogenic bacteria, viruses, etc.At the same time, in order to ensure the quality of food hygiene and protect the health and safety of Chinese consumers, the safety testing of livestock products must be strengthened.The traditional methods currently used in the detection of pig -affected infectious viruses are roughly: virus separation, electron microscope observation, poly...

Claims

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Application Information

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IPC IPC(8): C12Q1/70C12Q1/68C40B40/06C12R1/93
Inventor 穆祥董虹张涛姜代勋胡格段慧琴张岩王建舫李佳
Owner BEIJING UNIV OF AGRI
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