Sarkar F H, Crissman J D
Department of Pathology, Henry Ford Hospital, Detroit, MI 48202.
Biotechniques. 1990 Aug;9(2):180-5.
We have developed a polymerase chain reaction-based procedure for reproducible detection of the E6-E7 gene in human papilloma virus DNA sequences using formalin-fixed, paraffin-embedded tissue sections. This procedure is a simple one-step procedure which does not require any elaborate hybridization following polymerase chain reaction amplification. The protocol combines modified tissue treatment and proper primer selection for efficient amplification of target DNA in a highly specific manner allowing identification in ethidium bromide-stained gels. The procedure described here is useful for a variety of tissue preparations, particularly formalin-fixed, paraffin-embedded archival tissues.
我们已经开发出一种基于聚合酶链反应的程序,用于使用福尔马林固定、石蜡包埋的组织切片可重复地检测人乳头瘤病毒DNA序列中的E6-E7基因。该程序是一个简单的一步程序,在聚合酶链反应扩增后不需要任何复杂的杂交。该方案结合了改良的组织处理和合适引物的选择,以高度特异性的方式有效扩增靶DNA,从而可在溴化乙锭染色的凝胶中进行鉴定。这里描述的程序适用于多种组织制剂,特别是福尔马林固定、石蜡包埋的存档组织。