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Goose-origin kidney-type astrovirus GoAstV rapid diagnosis primer probe group, detecting method and application

A primer probe, astrovirus technology, applied in biochemical equipment and methods, microbial measurement/testing, recombinant DNA technology, etc., can solve the problem that it is not suitable for clinical detection of a large number of samples, the reproductive ability of goose embryos is not high, and it is not suitable for Clinical use and other issues, to achieve rapid differential diagnosis, high detection sensitivity, and reduce the cost of synthesis

Inactive Publication Date: 2018-11-16
POULTRY INST SHANDONG ACADEMY OF AGRI SCI SHANDONG SPECIFIC PATHOGEN FREEN CHICKS RES CENT
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Regarding the diagnosis of the virus, the traditional isolation and identification of the virus is time-consuming and laborious, and is not suitable for clinical use. Moreover, the virus has no hemagglutination, does not grow on chicken or duck embryos, and has low reproductive ability on goose embryos.
At present, the most common method is to use ordinary RT-PCR method to detect GoAstV, but this method is easy to cause cross-contamination between specimens, and gel electrophoresis is required for judgment, and it is not suitable for detection of a large number of clinical samples.

Method used

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  • Goose-origin kidney-type astrovirus GoAstV rapid diagnosis primer probe group, detecting method and application
  • Goose-origin kidney-type astrovirus GoAstV rapid diagnosis primer probe group, detecting method and application
  • Goose-origin kidney-type astrovirus GoAstV rapid diagnosis primer probe group, detecting method and application

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Embodiment 1

[0030] 1. Materials and methods

[0031] 1.1 Materials: The strain used in the test: GoAstV representative strain SD18, which was isolated and identified by our center.

[0032] 1.2 Method

[0033] 1.2.1 Design and synthesis of probes and primers:

[0034] According to the GoAstV 1b protein sequence (Genbank No: MF576430), primers and TaqMan MGB probes were designed using Primer ExPress3.0 software;

[0035] The sequence of the probe primer set is:

[0036] GoAstV-F: 5'TGGTGGTGGTGCGGTTTT 3';

[0037] GoAstV-R: 5' GGGCAACGTACCATCATAACG 3';

[0038] Probe: 5' FAM TGTAGAGACGGACTGGAC MGB 3';

[0039] The TaqMan MGB probe is labeled with a fluorophore at the 5' end and a quencher fluorophore at the 3' end;

[0040] The fluorescent group is FAM, and the quenched fluorescent group is MGB.

[0041] 1.2.2 Extraction of viral RNA

[0042] The RNA extraction of GoAstV strain SD18 was carried out according to Bioer’s Simple RNA kit operation, and the extracted RNA was used immedia...

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Abstract

The invention belongs to the technical field of the gene detection, and particularly relates to a goose-origin kidney-type astrovirus GoAstV rapid diagnosis primer probe group, a detecting method andapplication. The goose-origin kidney-type astrovirus GoAstV rapid diagnosis primer probe group comprises probe primer groups in the following sequences: GoAstV-F: 5' TGGTGGTGGTGCGGTTTT 3'; GoAstV-R: 5' GGGCAACGTACCATCATAACG 3'; and a TaqMan MGB probe: 5' FAM TGTAGAGACGGACTGGAC MGB 3'. The 5' end is marked as a fluorescence group by the probe, and the 3' end is marked as a quenching fluorescence group. The provided primer probe group has the advantages of higher sensibility, specificity, good repeatability, rapidness, high flux, less pollution and the like, and is used as a useful and powerfultechnology in the fields of a fundamental research, an application research and the like.

Description

technical field [0001] The invention belongs to the technical field of gene detection, and in particular relates to a rapid diagnosis primer probe set, detection method and application of goose-derived renal astrovirus GoAstV. Background technique [0002] Astrovirus (Astrovirus) is a single-stranded positive-sense RNA virus belonging to the family Astroviridae, with a genome length ranging from 6.4 to 7.9 kb. Astroviridae can be divided into mammalian genera and avian genera according to different hosts, among which mammalian genera can be divided into human astrovirus, feline astrovirus, porcine astrovirus, sheep astrovirus and mink astrovirus according to different hosts. Astroviruses; the avian Astrovirus genus includes duck astrovirus (DAstV), turkey astrovirus (TAstV), avian nephritis virus (ANV) and chicken astrovirus (CAstV). Since the second half of 2017, a new type of goose-derived astrovirus has been isolated from goose disease materials in Shandong area. Geneti...

Claims

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Application Information

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IPC IPC(8): C12Q1/70C12Q1/6851C12N15/11
CPCC12Q1/6851C12Q1/701C12Q2600/166C12Q2563/107C12Q2531/113C12Q2545/113
Inventor 袁小远孟凯王友令张玉霞艾武亓丽红
Owner POULTRY INST SHANDONG ACADEMY OF AGRI SCI SHANDONG SPECIFIC PATHOGEN FREEN CHICKS RES CENT
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