3种方法检测禽网状内皮组织增殖病毒人工感染鸡胚孵化雏鸡的带毒比较  

Comparison of Three Methods for Detecting the Viral Load in Chicken-embryo Chicks Artificially Infected with REV

作　　者：徐凤霞[1] 孙万里 张亚文 常爽[1] 王一新[1] 赵鹏[1] Xu Fengxia;Sun Wanli;Zhang Yawen;Chang Shuang;Wang Yixin;Zhao Peng(College of Animal Science and Veterinary Medicine,Shandong Agricultural University,Tai'an 271018,Shandong,China)

机构地区：[1]山东农业大学动物科技学院,山东泰安271018

出　　处：《中国动物检疫》2024年第2期80-85,共6页China Animal Health Inspection

基　　金：山东省重点研发计划(乡村振兴科技创新提振行动计划)项目(2022TZXD0019)。

摘　　要：为比较血液和粪便等样品采用不同方法检测禽网状内皮组织增殖病毒(REV)的准确率,从而为制定禽网状内皮组织增殖病(RE)净化程序提供参考,利用鸡胚卵黄囊接种构建REV垂直传播感染雏鸡模型,分别采用DF-1细胞病毒分离、RT-PCR检测和荧光定量RT-PCR检测,对1日龄雏鸡血液和胎粪样品,7~70日龄鸡(每隔1周)的血液和泄殖腔棉拭子样品,进行REV检测和结果比较。结果显示:对1日龄出壳的SPF鸡血浆进行检测,荧光定量RT-PCR灵敏度最高,阳性检出率为100%(25/25),其次为病毒分离,阳性检出率为96%(24/25),RT-PCR检出率最低;对1日龄出壳的SPF鸡胎粪进行检测,荧光定量RT-PCR与病毒分离灵敏度一致,阳性检出率均为80%(20/25),RT-PCR检出率最低,为28%(7/25)。对7~70日龄鸡(每隔1周)进行血液和泄殖腔棉拭子检测,3种方法阳性检出率与1日龄检测情况总体相似,检出率从高到低依次为荧光定量RT-PCR、病毒分离和RT-PCR。结果表明,在同一检测时间点使用同一检测方法,胎粪/泄殖腔棉拭子REV阳性检出率基本低于血液检出率。本研究初步比较了不同方法检测REV人工感染鸡胚孵化雏鸡的带毒情况,为实施REV净化奠定了基础。In order to compare the accuracy of various methods for detecting avian reticuloendotheliosis virus(REV)in blood and fecal samples and thus to provide a reference for the development of purification procedures for avian reticuloendotheliosis(RE),a model for REV vertical transmission to chicks was constructed using chickembryoyolksac inoculation.Blood and meconium samples from 1-day-old chicks,blood and cloacal swabs from 7 to 70-day-old chicks(every other week)were detected by RT-PCR,fluorescence quantitative RT-PCR and virus isolation method using DF-1 cells,respectively,and the results from which were compared.The results revealed that,for the plasma from 1-day-old SPF chicks,fluorescence quantitative RT-PCR had the highest sensitivity and positive detection rate of 100%,followed by virus isolation mehtod with the positive detection rate of 96%(24/25),and the last one was RT-PCR;for the meconium samples of 1-day-old SPF chicks,fluorescence quantitative RT-PCR and virus isolation shared the same sensitivity with the positive detection rate of 80%(20/25),and RT-PCR had the lowest detection rate of 28%(7/25).For the blood and cloacal swabs from 7 to 70-day-old chicks(every other week),the detection rates by the three methods were generally similar to those of 1-day-old chicks,that was,fluorescence quantitative RT-PCR,virus isolation and RT-PCR in successive order from high to low.In conclusion,the positive detection rate of REV in meconium/cloacal swabs was generally lower than that in blood using the same method for detecting samples within the same period.The methods for detecting the viral load of chicken-embryo chicks artificially infected with REV were preliminarily compared in the study,laying a foundation for future implementation of REV purification.

关 键 词：禽网状内皮组织增殖病毒 病毒分离 RT-PCR 荧光定量RT-PCR 

分 类 号：S851.3[农业科学—预防兽医学]