摘要
用胶体金标记提纯后的兔抗鸭肝炎病毒(DHV)IgG,建立了一种以微孔滤膜为固相载体,以红色胶体金为标记物的检测DHV的斑点免疫金渗滤法(DIGFA),并确定了最佳试验条件。该法既可定性,亦可定量,对纯化DHV的最小检测量为4.12ng/点,其敏感性为抗原斑点试验(AST)的2倍。特异性阻断试验和交叉反应试验证明DIGFA检测DHV具有较高的特异性。对DHV鸡胚尿囊液DIGFA法检出率为100%。DIGFA和AST法对36份临床样本检测阳性率分别为83.3%和75%,其阳性符合率为86.7%。随机抽样的6份DIGFA阳性肝脏样本经人工发病试验后电镜检查,均发现大量直径30~40nm的病毒粒子。实验结果表明,DIGFA法是一种微量、敏感、特异、快速、简便的检测DHV方法。
The purified rabbit anti-DHV IgG was labelled with the prepared colloidal gold. DIGFA with a microfilter paper as carrier and the red colloidal gold as label was developed for detection of duck hepatitis virus(DHV).The optimum working concentration of DHV antigen and rabbit anti-DHV IgG labelled with colloidal gold were determined to be 1∶320 and 1 ∶4.The detection of DIGFA had the advantages of qualitaive and quantitative analysis.The minimum amount of purified DHV antigen by the DIGFA was 4.12ng per dot,with the sensitivity of 2-fold more than the antigen spot test(AST).The high specificity of the DIGFA was shown by the specific blocking test of anti-DHV serum and the cross-rection test with DPV,GPV,NDV,IBDV,MDV,EDS76V and IBV.The positivc rate for allantoic fluid of experimentally inoculatcd embryonated eggs was 100%,The positive rate of 36 clinical duckliver samples measured by the DIGFA and AST were 83.3%and 75%,respectively. The agreement rate of DHV detected by the DIGFA and AST was 86.7%.Many virus particles were observed about 30 ̄40nm diameter by electromicroscope from specimens which were obtained by experimental infection with 6 random DIGFA-positive duck liver asmples. This study indicated that the DIGFA was a simple,trace,rapid,sensitive and specific method for the detection of DHV antigen.
出处
《安徽农业科学》
CAS
1997年第4期373-376,共4页
Journal of Anhui Agricultural Sciences
基金
农业部"九五"畜牧重点项目
关键词
鸭病
肝炎病毒
斑点免疫金渗滤
抗原斑点试验
Duck Hepatitis Virus(DHV),Dot Immuno-gold Filtration Assay(DIGFA),Antigen Spot Test(AST)