Brukner Ivan, El-Ramahi Razan, Sawicki Jacob, Gorska-Flipot Izabella, Krajinovic Maja, Labuda Damian
Centre de Recherche, Hôpital Sainte-Justine, Université de Montréal, 3175 Côte Sainte-Catherine, Montréal, Que, Canada.
J Clin Virol. 2007 Jun;39(2):113-8. doi: 10.1016/j.jcv.2007.03.013. Epub 2007 May 7.
Human papillomavirus (HPV) infection with oncogenic types is a prerequisite for cervical cancer development. HPV typing is required in the management of pre-cancerous lesions, epidemiological studies, and vaccination trials. None of the available HPV assays are satisfactory for routine diagnosis.
In order to develop an assay for clinically relevant HPV types, we generated HPV probes using in vitro selection scheme of iterative hybridization.
Starting from a mixture of random oligonucleotides, through several rounds of hybridization with 39 type-specific GP5+/6+ L1 sequences, we aimed to obtain specific probes to discriminate between these HPV types.
In vitro selection led to pools of specific probes, from which individual probes were cloned and tested for their diagnostic performance at ambient temperature. Typically, 10-fold stronger hybridization signals were obtained between each of the selected probes and their specific targets compared to signals with the remaining 38 HPV types. High sensitivity and specificity of selected probes was demonstrated a series of clinical samples in the hybridization assay.
A new panel of probes for detecting HPV types is described. Probes can be adapted for use in a simple clinical setting, or incorporated into different detection systems.
致癌型人乳头瘤病毒（HPV）感染是宫颈癌发生的必要条件。在癌前病变管理、流行病学研究及疫苗试验中，HPV分型是必需的。现有的HPV检测方法均无法令人满意地用于常规诊断。
为开发一种针对临床相关HPV类型的检测方法，我们采用迭代杂交的体外筛选方案生成了HPV探针。
从随机寡核苷酸混合物开始，通过与39种型特异性GP5+/6+ L1序列进行多轮杂交，我们旨在获得能区分这些HPV类型的特异性探针。
体外筛选产生了特异性探针库，从中克隆出单个探针并在常温下测试其诊断性能。通常，与其余38种HPV类型相比，每种选定探针与其特异性靶标之间的杂交信号要强10倍。在杂交试验中，一系列临床样本证明了选定探针具有高灵敏度和特异性。
描述了一组用于检测HPV类型的新探针。这些探针可适用于简单的临床环境，或整合到不同的检测系统中。
