利用聚合酶链反应对宫颈癌中人乳头瘤病毒16、18和33型转化基因区域进行扩增及特异性检测。
Amplification and specific detection of transforming gene region of human papillomavirus 16, 18 and 33 in cervical carcinoma by means of the polymerase chain reaction.
作者信息
Shimada M, Fukushima M, Mukai H, Kato I, Nishikawa A, Fujinaga K
机构信息
Department of Molecular Biology, Cancer Research Institute, Sapporo.
出版信息
Jpn J Cancer Res. 1990 Jan;81(1):1-5. doi: 10.1111/j.1349-7006.1990.tb02498.x.
Abstract
We have established a highly sensitive method for specific detection of human papillomavirus (HPV) 16, 18 and 33, by using the polymerase chain reaction (PCR). A HPV-related sequence (140 bp) in the E6 transforming region was specifically amplified and detected by gel electrophoresis and by the use of a specific oligonucleotide probe. The PCR could detect 10(5)-10(6) copies per cell (maximum sensitivity). Furthermore, HPV 16, 18 and 33 DNAs were synthesized in a common reaction solution and specifically detected by HPV type-specific probes. The PCR detected the HPV sequence from tissues which were negative to Southern hybridization. This detection technique may contribute significantly to the precise analysis of HPV in small proliferative lesions in the cervix.
摘要
我们通过聚合酶链反应（PCR）建立了一种用于特异性检测人乳头瘤病毒（HPV）16、18和33型的高灵敏度方法。E6转化区中一段与HPV相关的序列（140碱基对）通过凝胶电泳并使用特异性寡核苷酸探针进行特异性扩增和检测。PCR能够检测到每个细胞10⁵ - 10⁶个拷贝（最大灵敏度）。此外，HPV 16、18和33型DNA在同一反应溶液中合成，并通过HPV型特异性探针进行特异性检测。PCR能从Southern杂交呈阴性的组织中检测到HPV序列。这种检测技术可能对精确分析子宫颈小增殖性病变中的HPV有显著贡献。
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