摘要
将IBVT株基因组S1 基因上0.6kb 片段(位于S1 基因上1121bp～1723bp 之间) 克隆到PIBPCRCloning vector 中,用地高辛标记探针,分别与9 株IBV的PCR 产物和12 株IBV 的核酸进行点杂交均呈阳性反应,而该探针不与IBDV和NDV 的RNA,EDS76 病毒的DNA及正常鸡胚尿囊液的抽提物反应。探针检测IBVPCR产物的灵敏度为100～200pg。用该探针检测不同毒株IBV在鸡胚中的繁殖动态,接种24～30 小时后的的尿囊液均获阳性结果。用M 和宜株混合感染5 日龄雏鸡12 小时能从气管粘液中检出病毒,38～46 小时后能从肾组织中检出病毒。对7 只就诊病鸡的病变肾脏进行杂交检测,有5 只呈阳性反应。核酸杂交技术为生产上提供了直接从尿囊液和感染鸡组织中快速检测病毒,诊断IBV
The cloned 0.6kb sequence of s 1 gene of IBV T strain (The DNA sequences is located at positions of s 1 genes 1121bp to 1723bp)were labeled by digoxigenin as DNA Probe.The dot hybridizations were done for detection of IBV.All the RNA of 12 strains IBV and 0.6kb PCR sequences of 9 strain IBV were positive, but necleotide extracted from IBDV,NDV,EDS_76 and allantoic fluid of the uninoculated embryonating eggs were negative. As little as 200 pg and 100 pg of known positive target DNA (PCR products of IBV)could be detected with the two digoxigenin-labeled probes.H 120 ,M 41 and Yi strains were first detected with cloned digoxigenin labeled probe by 24～30 hours after inoculating.The viral genome of IBV was able to be detected using digoxigenin labeled probe early to 12 hours,36 hours respectively from the tracheal fluid and kidney tissues taken from chickens that were experimenally inoculated with the IBV M 41 strain and Yi strain.Dot hybridization were also used to detect the viral genome of IBV from 7 chickens nephritic with nephritis lesion.The viral genome from 5 chickens was positive.The nucleic acid hybridization could be used to detect the IBV from the allantoic fluid of infected embronating eggs and tissues of infected chickens.
出处
《中国预防兽医学报》
CSCD
2000年第1期64-66,共3页
Chinese Journal of Preventive Veterinary Medicine
关键词
支气管炎病毒
地高辛标记探针
点杂交
鸡
IBV
Chicken infectious bronchitis virus(IBV)
Digoxigenin labeled probe
Dot hybridization
