鸡源鹦鹉热衣原体MOMP基因重组腺病毒的稳定性及SPF雏鸡免疫试验

Stability test and SPF chick immune test of recombinant adenovirus containing major outer membrane protein of Chlamydia psittaci of chicken origin

目的:检测重组腺病毒质粒在HEK293细胞中的稳定性;重组腺病毒是否与减蛋综合征阳性血清发生中和反应,以及用重组腺病毒免疫SPF小鸡是否产生保护.方法:将重组腺病毒质粒在HEK293细胞中连续传代,并用PCR法检测目的基因;用减蛋综合征阳性血清在体外中和重组腺病毒,接种HEK293细胞;用重组腺病毒免疫SPF小鸡,然后用CpL菌株攻击.结果:重组腺病毒质粒在HEK293细胞中连续传代后,仍然用PCR检测能检测出目的基因;重组腺病毒与减蛋综合征阳性血清之间无交叉反应;用重组腺病毒免疫SPF鸡,用CpL菌株攻击,90%的小鸡获得了保护.结论:重组腺病毒质粒在HEK293细胞中能稳定传代;重组腺病毒不受减蛋综合征阳性血清的影响;用重组腺病毒免疫SPF小鸡,能够获得保护.

AIM： To detect the stability of the recombinant adenovirus plasmid containing major outer membrane protein （MOMP） gene from Chlamydla psittaci in HEK293 cells, to evaluate whether cross reactivity exists or not between recombinant adenovirus and positive serum of Egg Drop Symdrome （EDS）, and to evaluate whether SPF chicks vaccinated with the recombinant adenovirus were protected or not when challenged with virulent CpL strain. METHODS： The recombinant adenovirus plasmids were passaged in HEK293 cells in series and the target gene was detected by PCR. The recombinant adenovirus was mixed with positive serum of EDS in vitro and then infected HEK293 cells. The SPF chicks were vaccinated with recombinant adenovirus and then challenged with the CpL strain. RESULTS： The target gene of recombinant adenovirus plasmid could be amplified by PCR in different passages of HEK293 cells. There was no cross reactivity between recombinant adenovirus and positive serum of EDS. 90% of the SPF chicks vaccinated with recombinant adenovirus were protected when challenged with the CpL strain. CONCLUSION： The recombinant adenovirus plasmid can be passaged stably in HEK293 cells. The recombinant adenovirus can not react with positive serum of EDS. The SPF chicks inoculated with the recombinant adenovirus vaccination are protected.