Chimeric strain of porcine reproductive and respiratory syndrome virus and application thereof

A technology for respiratory syndrome and swine reproduction, applied in the directions of antiviral agents, viruses/phages, and viral antigen components, which can solve the problems of poor cross-protection of different subtype strains and limited immune protection of NADC30-like strains. Good immune protection, low toxicity

Active Publication Date: 2019-05-17
NANJING AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, commercial PRRS vaccines mainly include attenuated live vaccines and inactivated vaccines. Attenuated vaccines have better immune efficacy than inactivated vaccines, but have poor cross-protection against different subtypes of strains.
For example, attenuated vaccines such as TJM-F92, HuN4-F112, JXA1-R and R98 can effectively alleviate the clinical symptoms caused by HP-PRRSV and reduce piglet mortality, but their immune protection against NADC30-like strains is limited, so new broad-spectrum vaccines are being studied very necessary

Method used

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  • Chimeric strain of porcine reproductive and respiratory syndrome virus and application thereof
  • Chimeric strain of porcine reproductive and respiratory syndrome virus and application thereof
  • Chimeric strain of porcine reproductive and respiratory syndrome virus and application thereof

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Embodiment

[0042] 1 Materials and methods

[0043] 1.1 Viruses, cells and plasmids

[0044] The highly pathogenic strain BB0907 of PRRSV and the NADC30-like strain FJ1402 were isolated, identified and preserved by our laboratory. The infectious cDNA clone plasmid pCMV-TJM of the strain was constructed and preserved by our laboratory. Porcine primary alveolar macrophages (PAMs) were prepared according to the methods in the literature; Marc-145 cells were cryopreserved in liquid nitrogen in our laboratory and purchased from ATCC in the United States.

[0045] 1.2 Other reagents and experimental animals

[0046] RNA extraction kit Total RNA Kit I was purchased from OMEGA; SuperScript TM IIIReverse Transcriptase was purchased from Invitrogen; restriction enzymes AscI and SpeI were purchased from NEB; DMEM / F12 medium was purchased from Vicente; low melting point agar (Agarose) was purchased from SIGMA; IDEXX HerdChek PRRS X3ELISA kit was purchased from IDEXX. 30-35-day-old PRRSV, PCV2, ...

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Abstract

The invention discloses a chimeric strain of a porcine reproductive and respiratory syndrome virus and application thereof. Based on a full-length infectious cDNA clone of an attenuated strain of highly pathogenic PRRSV TJM-F92 vaccine and a NADC30-like subtype strain FJ1402, a reverse genetic technology is utilized to construct the chimeric strain of the porcine reproductive and respiratory syndrome virus. It is verified by tests that the chimeric strain has a lower virus growth speed and virus titer on Marc-145 cells than the original strain TJM-F92, and the virulence is low. The chimeric strain has a good immunoprotective effect on HP-PRRSV strain BB0907 and NADC30-like strain FJ1402 and can provide effective cross-immunization protection against infections of highly pathogenic strainsand NADC30-like strains, which lays an important foundation for the development of new broad-spectrum vaccines against this disease.

Description

technical field [0001] The invention relates to a new chimeric strain of porcine reproductive and respiratory syndrome virus, and also relates to a construction method of the chimeric strain and its application in preparing candidate vaccines for preventing porcine reproductive and respiratory syndrome epidemic strains. Background technique [0002] Porcine reproductive and respiratory syndrome (PRRS) is caused by porcine reproductive and respiratory syndrome virus (porcine reproductive and respiratory syndrome virus, PRRSV), a viral disease that seriously endangers the pig industry. Its clinical symptoms Complex and varied, characterized by reproductive failure in sows or respiratory disease in herds of all ages. The virus was isolated for the first time in my country in 1996, and then gradually spread throughout the country. In the summer and autumn of 2006, the highly pathogenic PRRSV (HP-PRRSV) broke out, its main clinical features were high fever, high morbidity and mo...

Claims

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Application Information

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IPC IPC(8): C12N7/01A61K39/12A61P31/14
Inventor 姜平张乔亚张日腾白娟姜辰龙
Owner NANJING AGRICULTURAL UNIVERSITY
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