6种不同J亚群禽白血病病毒检测试剂盒的比较  

Comparison of Six Test Kits for Detecting Avian Leukosis Virus Subgroup J

作　　者：赵方钰 崔慧珍 王一新[1] 常爽[1] 任志浩[1] 崔文平 赵鹏[1] ZHAO Fangyu;CUI Huizhen;WANG Yixin;CHANG Shuang;REN Zhihao;CUI Wenping;ZHAO Peng(College of Veterinary Medicine,Shandong Agricultural University,Tai'an 271017,China)

机构地区：[1]山东农业大学动物医学院,山东泰安271017

出　　处：《中国兽医杂志》2025年第4期62-69,共8页Chinese Journal of Veterinary Medicine

基　　金：中央引导地方科技发展专项资金项目(YDZX2022127)。

摘　　要：为了比较6种不同J亚群禽白血病病毒(ALV-J)检测试剂盒的检测结果,以更好地实施禽白血病净化和鸡苗洁净度评估。本试验首先将ALV-J参考株以卵黄囊接种感染无特定病原体(SPF)鸡胚构建雏鸡携带ALV-J模型,对出壳雏鸡逐一采集胎粪和抗凝血,随后分别将胎粪滤液和血浆接种鸡胚成纤维传代(DF-1)细胞进行病毒培养,维持7 d时收取细胞上清,以A、B、C三家公司的ALV p27抗原酶联免疫吸附测定(ELISA)试剂盒以及商品化的磁微粒化学发光检测试剂盒、胶体金试纸条和实时荧光定量PCR检测试剂盒分别对胎粪、不同稀释度胎粪样品、胎粪病毒分离细胞上清和血浆病毒分离细胞上清进行检测。结果显示,6种不同试剂盒对胎粪、胎粪病毒分离细胞上清和血浆病毒分离细胞上清的检测结果判定总体一致,其中以磁微粒化学发光检测试剂盒读数判定最为直观,其阳性样品与阴性样品的读值区分明显、易于判断。对不同稀释度胎粪样品的检测结果显示,以A公司ALV p27抗原ELISA试剂盒、磁微粒化学发光检测试剂盒和实时荧光定量PCR检测试剂盒相对灵敏,可检出低滴度病原。结果表明,不同检测方法对ALV-J检测的整体判定差异不显著,但针对低滴度样品检测时不同方法的检出率有差异,需要根据情况选择方法。In order to compare the detection results of six different avian leukosis virus subgroup J(ALV-J)detection kits for better avian leukosis purification and the assessment of chicken embryo cleanliness,this study first constructed a chick model carrying ALV-J by inoculating ALV-J reference strain into the yolk sac of specific pathogen free(SPF)chicken embryos.The embryos were infected and chick feces and anticoagulated blood were collected from the newly hatched chicks.The fecal filtrates and plasma were then inoculated into chicken embryo fibroblast DF-1 cells for virus culture.After 7 days of incubation,cell supernatants were collected and tested using enzyme-linked immunosorbent assay(ELISA)kits for ALV p27 antigen from companies A,B,and C,along with commercial magnetic microparticle chemiluminescence detection kits,colloidal gold test strips,and real-time fluorescence quantitative PCR kits.The samples tested included feces,different dilution samples of feces,virus isolation cell supernatants from feces,and plasma virus isolation cell supernatants.The results showed that the detection results from the six different kits for feces,fetal feces virus isolation cell supernatant,and plasma virus isolation cell supernatant were generally consistent,with the magnetic microparticle chemiluminescence detection kit offering the most intuitive readings.The positive and negative sample values were easily distinguishable.For the different dilution samples of feces,the A company's ALV p27 antigen ELISA kit,the magnetic microparticle chemiluminescence detection kit,and the real-time fluorescence quantitative PCR kit demonstrated relatively high sensitivity,capable of detecting low titer pathogens.These findings indicate that while the overall judgment of ALV-J detection using different methods did not show significant differences,the detection rates varied for low titer samples.Therefore,the method should be chosen based on the specific situation.

关 键 词：J亚群禽白血病病毒(ALV-J) P27抗原 酶联免疫吸附测定(ELISA) 磁微粒化学发光检测试剂盒 胶体金试纸条 实时荧光定量PCR 

分 类 号：S855[农业科学—临床兽医学]