The immune response and protective efficacy of a potential DNA vaccine against virulent Pasteurella multocida.

IF 3.6 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Journal, genetic engineering & biotechnology Pub Date : 2021-05-31 DOI:10.1186/s43141-021-00180-9
Ahmed A M Yassein, Ayaat A Teleb, Gamal M Hassan, Zaki A El Fiky
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引用次数: 2

Abstract

Background: Pasteurella multocida is the main cause of several infections of farm animals, and the immunity gained from commercial vaccines is for the short term only and needs to be routinely administered, so work on new vaccines against virulent P. multocida is crucial.

Results: In this study, the OmpH gene was amplified from ten P. multocida strains, and the PCR products were sequenced and analyzed. The results of RFLP analysis of OmpH gene digested by MspI enzyme showed that all of ten strains examined possessed one restriction site and two fragments, 350 and 650 bp. The OmpH sequence of strain No. 10 was cloned into bacterial expression vector pUCP24, and the recombinant pUCP24-OmpH was expressed in E. coli DH5α. Serum samples obtained from the ELISA test from a group of vaccinated rats indicate that the antibodies were present at high titer in immunized rats and can be tested as a vaccine candidate with a challenge.

Conclusions: In rats infected with the DNA vaccine and inactivated vaccine, a significant increase in serum antibody levels was observed. In addition, the DNA vaccine provided the vaccinated rats with partial protection; however, the protective efficacy was greater than that offered by the live attenuated vaccine. This successful recombinant vaccine is immunogenic and may potentially be used as a vaccine in the future.

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一种潜在的DNA疫苗对多杀性巴氏杆菌的免疫反应和保护效果。
背景:多杀性巴氏杆菌是几种家畜感染的主要原因,而从商业疫苗中获得的免疫力仅是短期的,需要常规接种,因此研究针对强毒性多杀性巴氏杆菌的新疫苗至关重要。结果:本研究从10株多杀假单胞菌中扩增到了OmpH基因,并对PCR产物进行了测序和分析。MspI酶切OmpH基因的RFLP分析结果显示,10株菌株均具有1个限制性内切位点和2个片段,分别为350和650 bp。将10号菌株的OmpH序列克隆到细菌表达载体pUCP24中,重组pUCP24-OmpH在大肠杆菌DH5α中表达。从一组接种过疫苗的大鼠的ELISA试验中获得的血清样本表明,该抗体在接种过疫苗的大鼠中以高滴度存在,可以作为一种有挑战的候选疫苗进行测试。结论:DNA疫苗和灭活疫苗感染的大鼠血清抗体水平显著升高。此外,DNA疫苗对接种大鼠具有部分保护作用;但其保护效果优于减毒活疫苗。这种成功的重组疫苗具有免疫原性,将来可能用作疫苗。
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