Rowland G F, Engelbrecht D J, Pool E J, Schmollgruber E C, Thompson G J, van der Merwe K J
Bioclones (Pty) Ltd, c/o Department of Biochemistry, University of Stellenbosch, Republic of South Africa.
J Virol Methods. 1989 Sep;25(3):259-69. doi: 10.1016/0166-0934(89)90053-0.
An ELISA test system for detection of plant viruses in field samples is described, based on the unlabelled antibody method using a peroxidase-antiperoxidase (PAP) complex. Novel features of the system include the use of acid-treated naked bacteria as combined carrier-adjuvants for the production of rabbit antiviral antibodies, and the use of acid-treated chicken antibodies (IgY) for antigen trapping in the ELISA. Systems for detection of potato virus Y (PVY), potato leafroll virus (PLRV), grapevine fanleaf virus (GFV) and grapevine virus A (GVA) were developed and compared with conventional direct double antibody sandwich (DAS) systems in tests with both purified virus and field samples. The PAP systems offer improved sensitivity, no background problems in the outer rows of the microtitre plates and are much easier to visualize with the naked eye if no plate reader is available.
本文描述了一种基于使用过氧化物酶 - 抗过氧化物酶（PAP）复合物的未标记抗体法检测田间样本中植物病毒的ELISA检测系统。该系统的新特点包括使用酸处理的裸细菌作为联合载体佐剂来生产兔抗病毒抗体，以及在ELISA中使用酸处理的鸡抗体（IgY）进行抗原捕获。开发了用于检测马铃薯Y病毒（PVY）、马铃薯卷叶病毒（PLRV）、葡萄扇叶病毒（GFV）和葡萄病毒A（GVA）的系统，并在对纯化病毒和田间样本的检测中与传统的直接双抗体夹心（DAS）系统进行了比较。PAP系统具有更高的灵敏度，在微量滴定板的外周没有背景问题，并且如果没有酶标仪，用肉眼更容易观察到结果。
