Yassein Ahmed A M, Teleb Ayaat A, Hassan Gamal M, El Fiky Zaki A
Genetics Department, Faculty of Agriculture, Fayoum University, 63514, Fayoum, Egypt.
J Genet Eng Biotechnol. 2021 May 31;19(1):81. doi: 10.1186/s43141-021-00180-9.
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.
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.
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.
多杀性巴氏杆菌是家畜多种感染的主要病因,而从商业疫苗获得的免疫力仅为短期,且需要定期接种,因此研发针对强毒多杀性巴氏杆菌的新型疫苗至关重要。
在本研究中,从十株多杀性巴氏杆菌菌株中扩增出OmpH基因,并对PCR产物进行测序和分析。用MspI酶消化OmpH基因的RFLP分析结果表明,所检测的十株菌株均具有一个限制性酶切位点和两个片段,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。一组接种疫苗大鼠的ELISA试验血清样本表明,免疫大鼠体内抗体呈高滴度存在,可作为候选疫苗进行攻毒试验。
在感染DNA疫苗和灭活疫苗的大鼠中,观察到血清抗体水平显著升高。此外,DNA疫苗为接种疫苗的大鼠提供了部分保护;然而,其保护效力大于减毒活疫苗。这种成功的重组疫苗具有免疫原性,未来有可能用作疫苗。