Heino P, Hukkanen V, Arstila P
Department of Virology, University of Turku, Finland.
J Virol Methods. 1989 Dec;26(3):331-7. doi: 10.1016/0166-0934(89)90115-8.
The presence of human papillomavirus (HPV) nucleotide sequences in paraffin sections of genital biopsies was examined by in situ hybridization using non-isotopic, digoxigenin-labeled probes representing HPV types 11, 16 and 18. Digoxigenin-labeling of the probes was performed using DNA labeling and a commercially provided detection kit. Hybridization was performed under stringent conditions. The hybrids were detected by using anti-digoxigenin alkaline phosphatase conjugate and visualized with enzyme catalyzed color reaction. In situ hybridization with digoxigenin-labeled probes was a useful technique for identification of HPV infection. The results were compared with the results obtained with radiolabeled DNA probes. The sensitivity of the digoxigenin-labeled probes was equal to the sensitivity of the radiolabeled probes. The background with digoxigenin-labeled probes was very low. Using nonradioactive probes the localization of hybrids at the cellular level was better than 35S-labeled probes.
采用非同位素地高辛标记探针原位杂交技术,检测了代表11、16和18型人乳头瘤病毒(HPV)的核苷酸序列在生殖器活检石蜡切片中的存在情况。使用DNA标记和市售检测试剂盒对探针进行地高辛标记。在严格条件下进行杂交。通过使用抗地高辛碱性磷酸酶结合物检测杂交体,并通过酶催化显色反应使其可视化。用地高辛标记探针进行原位杂交是鉴定HPV感染的一种有用技术。将结果与用放射性标记DNA探针获得的结果进行比较。地高辛标记探针的灵敏度与放射性标记探针的灵敏度相当。地高辛标记探针的背景非常低。使用非放射性探针时,杂交体在细胞水平的定位优于35S标记探针。