鸭瘟病毒疫苗株UL35缺失株的构建及其免疫保护效果评价

doi:10.11843/j.issn.0366-6964.2025.08.031

Abstract

Abstract:

This study aimed to study the biological characteristics and the immunoprotective effect of DEV vaccine strain with UL35 gene deletion. A UL35 gene-deletion virus strain was constructed based on the bacterial artificial chromosome (BAC) clone pDEV-EF1 which carries DEV vaccine strain full-length genome. The BAC clone pDEV-ΔVP26 of UL35 gene deletion was generated by two-step Red/ET recombination in E. coli. The recombinant virus rDEV-ΔVP26 was rescued from chicken embryo fibroblasts (CEFs) by calcium phosphate precipitation. And then the biological characteristics and immunoprotective capacity of recombinant virus were evaluated. Growth curves show that the virus titer of rDEV-ΔVP26 was stable increasing from 24 to 84 h, up to the peak value 10^5.36 TCID₅₀·0.1 mL^-1. During this period, the titer of rDEV-ΔVP26 was slightly reduced compared to the parental strain rDEV-EF1, and the plaque size of rDEV-ΔVP26 was decreased 10.60% compared to the parental virus. Animal experiments showed that 10⁶ TCID₅₀ rDEV-ΔVP26 is safe to 30-day-old ducks, and pre-immune ducks with 10⁵ TCID₅₀ rDEV-ΔVP26 could completely protect ducks from high virulent DEV challenge, this is similar to the rDEV-EF1 control group. The results showed that UL35 was non-essential gene of DEV, and absence of UL35 did not affect the immune protection capacity of DEV vaccine strain. These studies have laid a foundation for developing differential diagnosis vaccine between infected and vaccinated animals.

Key words: duck enteritis virus, VP26, nonessential gene, differential diagnosis

CLC Number:

S852.659.1

CHEN Liu, XIANG Shengrui, YUN Tao, NI Zheng, HUA Jionggang, ZHU Yinchu, ZHANG Cun, YE Weicheng. Construction and Evaluation of Immune Protection Capacity of a Recombinant Duck Enteritis Virus Deleting UL35 Gene[J]. Acta Veterinaria et Zootechnica Sinica, 2025, 56(8): 3933-3941.

Figures/Tables 8

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References 28

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引物名称Primers	序列Sequences
DEV-ΔVP26-for	5′- agagaacgctcttcatacaacaccacgtctacggcataacaacaaataaatgagtatgattcaata AGGATGACGACGATAAGTAGGG-3′
DEV-ΔVP26-rev	5′-aataaaagcttttatgaatctttattgaatcatactcatttatttgttgttatgccgtagacgtggCAA CCAATTAACCAATTCTGATTAG-3′
ΔVP26-JD-F	5′-gtgtgttttgtacgtatgccttgtg-3′
ΔVP26-JD-R	5′-gacgttcgcacatcaataatagct-3′

组别 Group	接种毒株 Virus	接种剂量/体积 Inoculation dose/volume	发病率/% Morbidity	死亡率/% Mortality
1缺失株Missing strain	rDEV-ΔVP26	10⁶ TCID₅₀/ 1 mL	0	0
2亲本毒株Parent strain	rDEV-EF1	10⁶ TCID₅₀/ 1 mL	0	0
3缺失株Missing strain	rDEV-ΔVP26	10⁵ TCID₅₀/ 1 mL	0	0
4亲本毒株Parent strain	rDEV-EF1	10⁵ TCID₅₀/ 1 mL	0	0
5对照Control	DMEM	1 mL	0	0

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Construction and Evaluation of Immune Protection Capacity of a Recombinant Duck Enteritis Virus Deleting UL35 Gene

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