不同禽胚和接种途径分离GoAstV JL01株的效果研究  

Isolation effects of GoAstV JL01 using different avian embryos and inoculation routes

作　　者：张雪 张玺 王文月 杨小艺 王尧 张亚如 门嘉琪 李洪涛 马波[1,2] 王君伟 ZHANG Xue;ZHANG Xi;WANG Wenyue;YANG Xiaoyi;WANG Yao;ZHANG Yaru;MEN Jiaqi;LI Hongtao;MA Bo;WANG Junwei(School of Veterinary Medicine,Northeast Agricultural University,Harbin 150030,China;School of Basic Medicine,Youjiang Medical University for Nationalities,Baise Guangxi 533000,China)

机构地区：[1]东北农业大学动物医学学院,哈尔滨150030 [2]右江民族医学院基础医学院,广西百色533000

出　　处：《东北农业大学学报》2024年第3期41-51,共11页Journal of Northeast Agricultural University

基　　金：国家自然科学基金项目(31502049)。

摘　　要：鹅星状病毒(Goose astrovirus,GoAstV)可导致1~15日龄雏鹅痛风甚至死亡。为探究有效分离GoAstV方法,将RT-PCR鉴定为GoAstV阳性的雏鹅肝脏、肾脏研磨液通过尿囊腔、绒毛尿囊膜及卵黄囊途径分别接种至SPF鸡胚、SPF鸭胚及非SPF鹅胚,通过多次传代观察胚体剖检变化、死亡情况、核酸、病毒载量检测等,综合判断GoAstV JL01株在禽胚上的最佳分离方式。结果表明,病料经尿囊腔途径接种非SPF鹅胚及SPF鸭胚,绒毛尿囊膜途径接种非SPF鹅胚、SPF鸡胚及SPF鸭胚,卵黄囊途径接种SPF鸭胚均无法使胚体发生显著的剖检变化及规律性死亡,RT-PCR及RT-qPCR检测表明,病毒无法在相应胚体、尿囊液及胚体其他组织中有效增殖,卵黄囊途径接种SPF鸡胚所传的10代胚体均发生发育迟缓、出血等特征性病变,除F_(1)代其余9代均死亡,卵黄囊和胚体中病毒载量为10^(4.5)~10^(6.0)copies·μL^(-1)。因此,卵黄囊途径接种8日龄SPF鸡胚为GoAstV JL01株最适分离方式。研究为GoAstV分离提供可借鉴路径。Goose astrovirus(GoAstV)can cause gout and even death in goslings aged between 1 to 15 days.In order to explore effective methods for isolating GoAstV,the homogenized samples of gosling liver and kidney identified as GoAstV positive by RT-PCR were inoculated into SPF chicken embryos,SPF duck embryos and non-SPF goose astrovirus by the embryo yolk sac(YA),chorioallantoic membrane(CAM)and chorioallantoic antrum(CAA)routes.The optimal method for isolating GoAstV JL01 on different avian embryos would be determined by comprehensive analyzing the results of the pathological changes in the embryos,mortality rates,and the detection of nucleic acid and viral loads after multiple passages.The results indicated that inoculation through the CAA route to non-SPF goose astrovirus and SPF chicken embryos,the CAM route to non-SPF goose astrovirus,SPF chicken embryos,and SPF duck embryos,and the YA route to SPF duck embryos did not lead to significant pathological changes or regular mortality in the embryos.RT-PCR and RT-qPCR tests revealed that the virus could not effectively replicate in the embryos,allantoic fluid or other tissues.However,when SPF chicken embryos were inoculated through the YA route and passed through 10 generations,all exhibited characteristic lesions such as retardation and hemorrhaging.All the generations except for the F_(1) died.The yolk sac and embryo tissues contained 10^(4.5)-10^(6.0) copies of goose astrovirus per 1μL of tissue cDNA.Therefore,inoculation via the YA route in 8-day-old SPF chicken embryos was identified as the optimal method for isolating GoAstV JL01.This study provided a reference pathway for the isolation of GoAstV.

关 键 词：鹅星状病毒 卵黄囊 鸡胚 分离 

分 类 号：S852.65[农业科学—基础兽医学]