Visual chip for synchronously detecting six avian viruses

A technology for avian virus and avian leukemia virus, which is applied in biochemical equipment and methods, resistance to vector-borne diseases, and determination/inspection of microorganisms, etc.

Active Publication Date: 2020-04-28
SICHUAN AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

This technology allows for accurate detection of different types of diseases caused by birds that are transmitted through droplets from other animals or humans during flight. It also includes methods like polymerase chain reaction (PCR) which have been previously used more efficiently than traditional techniques such as ELISA. Overall this new technique provides an efficient way to diagnose various bird conditions quickly and easily without expensive equipment.

Problems solved by technology

This patents describes various techniques such as diagnosing birds against these illnesses like classical cytology based on their ability to produce certain proteins called antibodies from cells within affected areas during fetal development stages. These technical problem addressed by current vaccine researchers include lack of efficient testing tools for identifying all three types of coronavirus including avian rickettsia, new castle disease virus type 1, and newly emerging strain associated with each etiological agent causing gastrointestinal tract damage leading to economic degradations worldwide. Current solutions involve expensive labor intensive procedures involving physical examination and live animal sacrifice.

Method used

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  • Visual chip for synchronously detecting six avian viruses
  • Visual chip for synchronously detecting six avian viruses
  • Visual chip for synchronously detecting six avian viruses

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0078] Embodiment 1 The preparation of visualized gene chip

[0079] 1. Probe Synthesis

[0080] Probes were synthesized according to Table 1, each with an amino group at the 5' end of the probe.

[0081] Table 1 Probe sequence list

[0082]

[0083]

[0084] 2. Array Design

[0085] According to the size of the aldehyde-based substrate and the number of probes, the gene chip array is designed, and each chip has 3 repeated arrays, and each repeated array is designed with 4 rows of probe genes. The positions of the probes of each viral target gene on the substrate are as follows: the first three in the first row are negative controls, the last three in the first row are positive controls (λ positioning genes), the first three in the second row are ALV, and the second three in the second row are positive controls. The last three in the third row are MDV, the first three in the third row are IBDV, the last three in the third row are NDV, the first three in the fourth row...

Embodiment 2

[0092] Example 2 Visualization gene chip detection

[0093] 1. Materials

[0094] Target gene recombination plasmid: T / AIV-NP, T / DNV-F, T / ILTV-TK, T / IBV-N; lambda positioning gene recombination plasmid; Escherichia coli DH5α; MDV, IBDV virus tissue fluid; ALV DNA nucleic acid.

[0095] 2 methods

[0096] 2.1 Design and synthesis of primers

[0097] Using Megaalign of DNASTAR software, the gene sequences of ALV, MDV and IBDV strains included in NCBI were compared and analyzed, and Primer5.0 was used to design 3 pairs of primers (env gene of ALV, meq gene of MDV, VP2 gene of IBDV). The gene amplification length is between 200 and 800bp, and the Tm is at (55±5)°C. The primers are synthesized by Beijing Qingke Xinye Biotechnology Co., Ltd. (see Table 2). Make touch ups.

[0098] Table 2 Target gene primers of ALV, MDV and IBDV

[0099]

[0100] 2.2 Amplification and identification of ALV-env, MDV-meq, and IBDV-VP2 target genes

[0101] Extract the total RNA of virus strai...

Embodiment 3

[0117] Example 3 Visual gene chip spraying times optimization

[0118] 1. Method

[0119] Add the oligonucleotide probe solution with a final concentration of 50p mol / L into the corresponding wells of the 384-well sampling plate, and add the solution with ddH 2 O served as a negative control. Using a chip spotter, according to step 2.5.4, spray samples on three aldehyde-based substrates for 1, 2, and 3 times in sequence, prepare chips with different spraying times, and seal and fix them. Subsequently, observe the results according to the steps of visual chip detection in Example 2.

[0120] 2. Results

[0121] The results showed that there was no obvious difference in the spot signal when the samples were sprayed once or twice, but when the samples were sprayed for the third time, the displacement between the probes caused the hybridization signals to be connected together, which affected the judgment of the final hybridization result. And as the number of spraying samples...

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Abstract

The invention provides a visual gold-labeled silver staining gene chip. The chip can be used for simultaneously detecting at least one of avian leukosis virus, Marek's disease virus, infectious bursaldisease virus, newcastle disease virus, avian influenza virus and infectious laryngotracheitis virus. The invention also provides a detection method of the avian leukosis virus, the Marek's disease virus, the infectious bursal disease virus, the newcastle disease virus, the avian influenza virus or the infectious laryngotracheitis virus. The method is realized by utilizing the chip. The method has the advantages of low cost, simplicity in operation and high flux, and has a wide application prospect in the field of avian virus detection.

Description

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Claims

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Application Information

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Owner SICHUAN AGRI UNIV
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