摘要
本研究首次报道应用银加强胶体金探针检测鸡减蛋综合征（ＥＤＳ７６）病毒获得成功。经初步提纯后的ＥＤＳ７６病毒免疫兔制备高免血清，按常规方法提取兔抗ＥＤＳ７６病毒的ＩｇＧ，成功地建立了检测ＥＤＳ７６病毒抗原的银加强胶体金探针法（ＳＥＣＧＰＡ），并确定了操作程序的最佳试验条件为：兔抗ＥＤＳＶＩｇＧ工作浓度为１４００，金标记羊抗兔ＩｇＧ的工作浓度为１２０，银染色的时间为１５ｍｉｎ。用该法对提纯的ＥＤＳ７６病毒（ＥＤＳＶ）抗原的最低检出量为０．１１７１９μｇ／ｍｌ。以此法检测了人工感染的５０只鸡采集的样本，对卵巢、输卵管峡部、咽喉部、软壳蛋的阳性率均为１００％，粪便阳性率为９２％，而检测鸡新城疫病毒、鸡传染性法氏囊病病毒、鸡传染性支气管炎病毒、鸡传染性喉气管炎病毒、大肠杆菌、巴氏杆菌、沙门氏菌等鸡源病毒和细菌样品时全部呈阴性反应。研究结果表明，该法具有快速、准确、操作简便、敏感性高、易于判读、特异性强、重复性好等优点。
A silver enhanced colloidal gold probe assay (SECGA) was developed for the detection of egg drop syndrome virus (EDSV) for the first time.The optimum working concentration of rabbit anti EDSV immunoglobulin G (IgG) was determined to be 1:400,that of sheep anti rabbit IgG labelled with colloidal gold to be 1∶20 and the optimum time of silver staining to be 15 minutes.The detection limit of purified EDSV antigen by the SECGA was 0.11719μg/ml.The high specificity of the SECGA was shown by the specific blocking test of rabbit anti EDSV serum and cross reaction test with Newcasstle Disease virus (NDV),Infectious Busal Disease Virus (IBDV),Infectious bronchitis Virus (IBV),Infectious Laryngotracheitis Virus (ILTV),Escherichia coli of chicken origin,Pasteurella multocida of chicken origin and Salmonella gallinarum.Specimens of the ovariums,tubules,throat and excrement from 50 egg laying chickens infected with EDSV were determined,the positive rate were 100%,100%,100%and 92%,respectively.This study indicates that the SECGA is a rapid,highly sensitive,highly specific and reproducible method for the detection of EDSV antigen.It is an efficient means for rapid diagnosis and epidemiological investigation of EDSV infection.
出处
《中国兽医杂志》
CAS
北大核心
1997年第7期9-11,共3页
Chinese Journal of Veterinary Medicine
基金
四川省教委资助的重点科研项目
关键词
检测
减蛋综合征
病毒
鸡病
SECGPA
egg drop syndrome virus (EDSV),Silver enhanced colloidal gold assay(SECGA)
