Gentilomi G, Zerbini M, Gallinella G, Venturoli S, Manaresi E, Morandi R, Musiani M
Department of Clinical and Experimental Medicine, University of Bologna, Italy.
Prenat Diagn. 1998 Apr;18(4):363-8.
In our study we describe the direct detection of parvovirus B19 capsid antigens in amniotic fluid samples for the rapid and simple prenatal diagnosis of B19 induced fetal hydrops. The assay was performed on amniotic fluid specimens from fetal hydrops dotted on nylon membranes. The two capsid antigens, VP1 and VP2, which represent four per cent and 96 per cent of the capsid, respectively, were detected using a pool of monoclonal antibodies directed against these two proteins and the complex was visualized by immunoperoxidase staining. The assay could be performed in about four hours and positive results were revealed at the end of the reaction as dark blue spots on the nylon membrane. We analysed 26 amniotic fluid samples from 26 selected cases of non-immune hydrops for the presence of B19 antigens. Out of these 26 samples, 13 had previously proved positive for B19 DNA, detected by dot blot hybridization and/or in situ hybridization and/or nested PCR, and 13 had proved negative. The results obtained with our assay were compared with results obtained for the presence of B19 DNA and a close agreement was found. The method is simple and rapid to perform, does not require costly instruments, and all the reagents used in the assay are commercially available. The assay described can thus be useful for a prompt counselling and management of B19 fetal infection.
在我们的研究中，我们描述了直接检测羊水样本中的细小病毒B19衣壳抗原，用于快速、简单地产前诊断由B19引起的胎儿水肿。该检测方法是对点滴在尼龙膜上的胎儿水肿羊水标本进行的。使用针对这两种蛋白质的单克隆抗体池检测分别占衣壳4%和96%的两种衣壳抗原VP1和VP2，并通过免疫过氧化物酶染色使复合物可视化。该检测大约可在4小时内完成，反应结束时阳性结果显示为尼龙膜上的深蓝色斑点。我们分析了26例选定的非免疫性水肿病例的羊水样本中是否存在B19抗原。在这26个样本中，13个样本先前经斑点印迹杂交和/或原位杂交和/或巢式PCR检测已证明B19 DNA呈阳性，13个样本已证明为阴性。将我们检测获得的结果与B19 DNA存在情况的检测结果进行比较，发现两者吻合度很高。该方法操作简单、快速，不需要昂贵的仪器，检测中使用的所有试剂均可从市场上购得。因此，所描述的检测方法有助于对B19胎儿感染进行及时的咨询和处理。
