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Primers for identifying avian infectious bronchitis virus strain type, RT-PCR detection kit, method and application

A technology of RT-PCR and bronchitis, which is applied in the field of typing and identification of common chicken infectious bronchitis virus vaccine strains, can solve the problems of time-consuming and laborious, and achieve the effect of low cost, strong specificity and high sensitivity

Active Publication Date: 2018-07-24
青岛嘉智生物技术有限公司 +2
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Virus serotyping requires neutralization experiments, which is time-consuming and labor-intensive

Method used

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  • Primers for identifying avian infectious bronchitis virus strain type, RT-PCR detection kit, method and application
  • Primers for identifying avian infectious bronchitis virus strain type, RT-PCR detection kit, method and application
  • Primers for identifying avian infectious bronchitis virus strain type, RT-PCR detection kit, method and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0057] Example 1 Primers

[0058] The embodiment of the present invention provides a kind of primer for identifying the strain type of chicken infectious bronchitis virus, including four pairs of typing primers, and the sequences of the primers are as follows:

[0059] Primer one:

[0060] H120 upstream primer: 5'-ATTGCTTACGGTCCTCT-3',

[0061] H120 downstream primer: 5'-CTGCTGGTTGACATCTT -3';

[0062] Primer 2:

[0063] QX upstream primer: 5'-GTACAGGGTCTTGTCCTA-3',

[0064] QX downstream primer: 5'-GTGTTGCTTAATTCACCT -3';

[0065] Primer three:

[0066] LDT3 upstream primer: 5'-CGCCACAGCAGGAAGAAT -3',

[0067] LDT3 downstream primer: 5'-GTCCGTAGTTGGAATGAAGA -3';

[0068] Primer four:

[0069] 4 / 91 upstream primer: 5'-CCAGATAGGCGGTGTTAG -3',

[0070] 4 / 91 downstream primer: 5'- TCGGCAATAGGAAAGTGT-3'.

Embodiment 2

[0071] Embodiment 2 detection reagent

[0072] The embodiment of the present invention provides a RT-PCR detection reagent for identifying the strain type of chicken infectious bronchitis virus, including the following components: RNA extraction reagent, reverse transcription reagent, PCR reaction reagent, four pairs of typing primers , positive control sample;

[0073] Wherein, the sequence of primer is as follows:

[0074] Primer one:

[0075] H120 upstream primer: 5'-ATTGCTTACGGTCCTCT-3',

[0076] H120 downstream primer: 5'-CTGCTGGTTGACATCTT -3';

[0077] Primer 2:

[0078] QX upstream primer: 5'-GTACAGGGTCTTGTCCTA-3',

[0079] QX downstream primer: 5'-GTGTTGCTTAATTCACCT -3';

[0080] Primer three:

[0081] LDT3 upstream primer: 5'-CGCCACAGCAGGAAGAAT -3',

[0082] LDT3 downstream primer: 5'-GTCCGTAGTTGGAATGAAGA -3';

[0083] Primer four:

[0084] 4 / 91 upstream primer: 5'-CCAGATAGGCGGTGTTAG -3',

[0085] 4 / 91 downstream primer: 5'- TCGGCAATAGGAAAGTGT-3'.

[0086]...

Embodiment 3

[0091] Example 3 Detection Kit

[0092] The embodiment of the present invention provides an RT-PCR detection kit for identifying the strain type of chicken infectious bronchitis virus, specifically the RT-PCR detection kit for identifying the strain type of chicken infectious bronchitis virus provided in Example 2. - A kit of PCR detection reagents.

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Abstract

The invention discloses primers for identifying an avian infectious bronchitis virus strain type, an RT-PCR detection kit, a method and application. Sequences of the primers are as follows: an H120 forward primer: 5'-ATTGCTTACGGTCCTCT-3', and an H120 reverse primer: 5'-CTGCTGGTTGACATCTT-3'; a QX forward primer: 5'-GTACAGGGTCTTGTCCTA-3', and a QX reverse primer: 5'-GTGTTGCTTAATTCACCT-3'; an LDT3 forward primer: 5'-CGCCACAGCAGGAAGAAT-3', and an LDT3 reverse primer: 5'-GTCCGTAGTTGGAATGAAGA-3'; a 4 / 91 forward primer: 5'-CCAGATAGGCGGTGTTAG-3', and a 4 / 91 reverse primer: 5'-TCGGCAATAGGAAAGTGT-3'. The primers, the RT-PCR detection kit, the method and the application have the beneficial effects that the kit provided by the invention separately amplifies four fragments of different sizes by only once gradient RT-PCR, and then, vaccine strains and popular wild strains of infectious bronchitis of chickens can be rapidly identified and diagnosed; the kit has the characteristics of high efficiency,low cost, high specificity, high sensitivity and the like; a convenient and quick detection method is provided for the differential diagnosis and molecular epidemiological analysis of avian infectious bronchitis viruses and prevention and control of the avian infectious bronchitis viruses.

Description

technical field [0001] The invention relates to the technical field of molecular biology detection, in particular to a primer for identifying the strain type of chicken infectious bronchitis virus, a RT-PCR detection kit and its application, which is applicable to common chicken infectious bronchitis virus vaccine viruses Type identification of strains. Background technique [0002] Avian infectious bronchitis is an acute, highly contagious respiratory disease that occurs only in chickens and is caused by avian infectious bronchitis virus with worldwide distribution. All ages are susceptible, young chickens are characterized by panting, and egg production and quality decline in laying hens. The prevalence of infectious bronchitis has caused huge economic losses to the poultry industry and seriously restricted the development of the poultry industry. [0003] Avian infectious bronchitis virus belongs to the Coronaviridae family, the genus Coronaviridae, and its genome has...

Claims

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Application Information

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IPC IPC(8): C12Q1/70C12Q1/686C12N15/11C12R1/93
CPCC12Q1/686C12Q1/701C12Q2565/125
Inventor 高绪慧秦立廷李志中魏笑笑吴海洋沈志勇刁秀国武梅
Owner 青岛嘉智生物技术有限公司
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