| 斑点免疫金渗滤法检测鸭肝炎病毒的研究 |
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| 引用本文： | 张小飞,陈毓川.斑点免疫金渗滤法检测鸭肝炎病毒的研究[J].安徽农业科学,1997,25(4):373-376. |
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| 作者姓名： | 张小飞 陈毓川 |
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| 作者单位： | 安徽省农科院畜牧兽医研究所 |
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| 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。
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| 关 键 词： | 鸭肝炎病毒（DHV） 斑点免疫金渗滤法（DIGFA） 抗原斑点试验（AST） |
Studies on the Detection for Duck Hepatitis Virus by Dot Immuno-gold Filtration Assay |
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| Abstract: | 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. |
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| Keywords: | Duck Hepatitis Virus(DHV) Dot Immuno-gold Filtration Assay(DIGFA) Antigen Spot Test(AST) |
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