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采用地高辛标记探针原位杂交法检测生殖器活检标本中的人乳头瘤病毒(HPV)DNA 。

Detection of human papilloma virus (HPV) DNA in genital biopsy specimens by in situ hybridization with digoxigenin-labeled probes.

作者信息

Heino P, Hukkanen V, Arstila P

机构信息

Department of Virology, University of Turku, Finland.

出版信息

J Virol Methods. 1989 Dec;26(3):331-7. doi: 10.1016/0166-0934(89)90115-8.

Abstract

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.

摘要

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。

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