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Antigen polypeptides of porcine reproductive and respiratory syndrome virus and application of antigen polypeptides

A technology for respiratory syndrome and antigenic peptides, applied in viral peptides, viruses, viruses/phages, etc., can solve problems such as inability to distinguish vaccine immunization from wild virus infection, difficulties in PRRS disease detection and prevention, and achieve good specificity.

Active Publication Date: 2021-08-31
HARBIN VETERINARY RES INST CHINESE ACADEMY OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The attenuated vaccine has played a positive role in preventing PRRS, but the existing serological differential diagnosis method cannot distinguish vaccine immunity from wild virus infection, which brings great difficulties to the detection and prevention of PRRS disease

Method used

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  • Antigen polypeptides of porcine reproductive and respiratory syndrome virus and application of antigen polypeptides
  • Antigen polypeptides of porcine reproductive and respiratory syndrome virus and application of antigen polypeptides
  • Antigen polypeptides of porcine reproductive and respiratory syndrome virus and application of antigen polypeptides

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Screening of Predominant Antigen Region of PRRSV NSP2

[0032] 1. Experimental materials

[0033] 1.1 Experimental serum

[0034] PRRSV HuN4-F112 positive sera, PRRSV HuN4-F5 positive sera, and PRRSV negative sera are preserved by our laboratory.

[0035] 1.2 Experimental reagents

[0036] Peptides were synthesized by Jill Biochemical Co., Ltd.; TMB chromogenic solution was purchased from Invitrogen; HRP goat anti-pig IgG was purchased from SIGMA; ELISA plate was purchased from Corning; PBS dry powder was purchased from Beijing Kulaibo Technology Co., Ltd.

[0037] 1.3 Experimental Instruments

[0038] The electric constant temperature incubator was purchased from Shanghai Yiheng Technology Co., Ltd.; the pH meter was purchased from; the oscillator was purchased from Qilin Bell Instrument Manufacturing Company; the plate washer was purchased from BioTek Company; the automatic microplate reader was purchased from BioTek Company; Instruments were purchased from Millip...

Embodiment 2

[0091] Establishment of ELISA method for porcine reproductive and respiratory syndrome deletion mutant strain

[0092] In this example, the deleted m1B and m2B polypeptides were used as coating antigens to establish an indirect ELISA method for detecting PRRSV deletion marker strains. Through optimization, the optimal reaction conditions for each step were determined. The optimal coating concentration of 1B2B antigen was 1.25 μg / ml, the optimal dilution of primary antibody serum was 1:40, and the optimal dilution of enzyme-labeled secondary antibody was 1:30000. The optimal incubation time for the primary antibody and the secondary antibody is 60 minutes, and the optimal color development condition for the substrate is 15 minutes at room temperature. The critical value of positive and negative is 0.237. The serum after HuN4-F112 immunization was detected by 1B2B-ELISA method, and the results showed that the specific antibody against 1B2B could be detected from the 21st day af...

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Abstract

The invention discloses a group of antigen polypeptides of porcine reproductive and respiratory syndrome virus and application of the antigen polypeptides, relates to the technical field of biological detection, and aims to solve the problem that PRRS genetic engineering marked attenuated vaccines which are immunized by vaccines and infected by wild viruses cannot be effectively distinguished in serology. The antigen peptide provided by the invention is a composition composed of one or two polypeptides as shown in SEQ ID NO: 1 and SEQ ID NO: 2 in a sequence table. The antigen peptide is used for preparing an ELISA diagnostic kit. Protein polypeptides which can be used for differential diagnosis are screened, and a corresponding indirect ELISA detection method is established. Candidate tag proteins are provided for marking attenuated vaccine strains through PRRS genetic engineering in the future, and technical support is provided for differential diagnosis of the PRRS vaccine strains and wild strains. The invention is applied to the field of pig vaccines and diagnosis.

Description

technical field [0001] The invention belongs to the technical field of biological detection, and in particular relates to a group of porcine reproductive and respiratory syndrome (PRRS) virus antigen polypeptides and a polypeptide ELISA diagnostic kit. Background technique [0002] Porcine reproductive and respiratory syndrome (PRRS) is a contagious viral disease of pigs, which has caused huge economic losses to the pig industry all over the world. It is also called "blue ear disease" (Keffaber et al., 1989) because the ears of affected pigs often turn blue and cyanotic. Clinically, it mainly manifests as reproductive disorder in pregnant sows and respiratory disorder in pigs of all ages. PRRS first appeared in the United States in 1987, and then the disease became widespread in the United States and spread to Canada and other regions. Subsequently, the disease was also reported in some countries and regions in Europe and Asia (Albina et al., 1997). In 1992, the World Org...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/08G01N33/569G01N33/543
CPCC07K14/005G01N33/56983G01N33/54306C12N2770/10022G01N2333/08
Inventor 安同庆蔡雪辉杨勇博王海伟
Owner HARBIN VETERINARY RES INST CHINESE ACADEMY OF AGRI SCI
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